WO2019183117A1 - Methods and compositions comprising genetically engineered vaccines for the treatment and prevention of cancer - Google Patents

Abstract

Provided herein are methods and compositions for treating cancer in an individual comprising administering to the individual an effective amount of a dendritic cell therapeutic vaccine incorporating the p53 tumor suppressor and administering at least one CD122/CD132 agonist and at least one immune checkpoint inhibitor to treat a cancer in a subject. Also provided herein are methods of enhancing anti-tumor efficacy by administering the agents described above in combination with other cancer therapies.

Description

METHODS AND COMPOSITIONS COMPRISING GENETICALLY

ENGINEERED VACCINES FOR THE TREATMENT AND PREVENTION OF

CANCER

[0001] This application claims the benefit of United States Provisional Patent Application No. 62/645,044, filed March 19, 2018, which is incorporated herein by reference in its entirety.

BACKGROUND

1. Field

[0002] The present invention relates generally to the fields of biology and medicine. More particularly, it concerns methods and compositions that utilize gene and cellular vaccines which induce anti-p53 immune responses for the treatment and prevention of cancer.

2. Description of Related Art

[0003] Current therapies for cancer involve locoregional treatments like surgery or radiation and systemically administered agents like chemotherapy and monoclonal antibodies. More recently, multiple immune therapies for cancer have been approved by regulatory agencies such as immune checkpoint inhibitors like Yervoy, Keytruda and Optivo; oncolytic viruses like TVEC; dendritic cell vaccines like Provenge; recombinant protein immune stimulatory cytokines like Roferon and Aldesleukin; and chimeric antigen receptor (CAR) T cell therapies like Kymriah (tisagenlecleucel) and Yescarta (axicabtagene ciloleucel). A plethora of immunotherapeutic agents are being evaluated as monotherapies and in combinations in numerous pre-clinical and clinical studies.

[0004] Despite these advances in cancer immunotherapy, it is generally appreciated that the majority of patients do not respond to these treatments and it is estimated there will be approximately 500,000 deaths from cancer yearly in the United States. Hence, there is an unmet need for improved cancer immune therapies which can increase their efficacy. SUMMARY

[0005] In certain embodiments, the present disclosure provides methods to treat cancer in a subject by administering an effective amount of a p53 dendritic cell vaccine and at least one preferential CD122/CD132 agonist to said subject. In some aspects, the p53 dendritic cell vaccine is further defined as a dendritic cell therapeutic vaccine incorporating the p53 tumor suppressor as an antigen to generate an anti-p53 immune response. In some aspects, the preferential CD122/CD132 agonist preferentially binds to the CD122/CD132 receptor complex and has lower affinity binding for the CD25 receptor and/or the IL-l5a receptor. In certain aspects, more than one preferential CD122/CD132 agonist is administered. In particular aspects, the subject is a human. In some aspects, the p53 dendritic cell vaccine is prepared using an adenoviral p53 vector.

[0006] In certain aspects, the CD122/CD132 agonist preferentially binds to the CD122/CD132 receptor complex and has lower affinity binding for CD25 or the IL15 alpha receptor as compared to the affinity binding to the CD 122/CD 132 receptor complex. In specific aspects, the one or more CD122/CD132 agonists are an IL-2/anti-IL-2 immune complex, an IL-l5/anti-IL-l5 immune complex, an IL-15/IL-15 Receptor a-IgGl-Fc (IL-l5/IL-l5Ra- IgGl-Fc) immune complex, PEGylated IL-2, PEGylated IL-15, IL-2 mutein and/or IL-15 mutein. The CD122/CD132 agonist may be an IL-15 mutant (e.g., IL-15N72D) bound to an IL-15 receptor a/IgGl Fc fusion protein, such as ALT-803. In certain aspects, IL-15 is pre- complexed with IL-15 receptor alpha (IL-l5Ra) to preferentially bind to CD122/CD132. In particular aspects, the IL-2 receptor agonist is not F42K.

[0007] In some aspects, the at least one preferential CD123/CD132 agonist is delivered at doses ranging between 5-100 ug/kg. In certain aspects, the at least one preferential CD123/CD32 agonist is administered subcutaneously (SQ) or intravenously (IV). In some aspects, the at least one preferential CD123/CD132 agonist is administered at intervals ranging from weekly to every 2-4 weeks per treatment cycle. In some aspects, more than one treatment cycle of the preferential CD 123/CD 132 agonist is given.

[0008] In some aspects, the method further comprises administering at least one additional anticancer treatment. In certain aspects, the at least one additional anticancer treatment is surgical therapy, chemotherapy, radiation therapy, hormonal therapy, immunotherapy, small molecule therapy, receptor kinase inhibitor therapy, anti- angiogenic therapy, cytokine therapy, cryotherapy, radioablation or a biological therapy. In some aspects, the biological therapy is a monoclonal antibody, siRNA, miRNA, antisense oligonucleotide, ribozyme, gene editing, cellular therapy or gene therapy.

[0009] In some aspects, the at least one additional anticancer treatment is an immune checkpoint inhibitor. In certain aspects the immune checkpoint inhibitor is of CTLA-4, PD-l, PD-L1, PD-L2, LAG-3, BTLA, B7H3, B7H4, TIM3, KIR, or A2aR. In some aspects, the at least one immune checkpoint inhibitor is an anti-CTLA-4 antibody. In some aspects, the anti- CTLA-4 antibody is tremelimumab or ipilimumab. In certain aspects, the at least one immune checkpoint inhibitor is an anti-killer-cell immunoglobulin-like receptor (KIR) antibody. In some embodiments, the anti-KIR antibody is lirilumab. In some aspects, the inhibitor of PD- Ll is durvalumab, atezolizumab, or avelumab. In some aspects, the inhibitor of PD-L2 is rHIgMl2B7. In some aspects, the LAG3 inhibitor is IMP321, or BMS-986016. In some aspects, the inhibitor of A2aR is PBF-509. In some aspects, the at least one immune checkpoint inhibitor is a human programmed cell death 1 (PD-l) axis binding antagonist. In certain aspects, the PD-l axis binding antagonist is selected from the group consisting of a PD-l binding antagonist, a PDL1 binding antagonist and a PDL2 binding antagonist. In some aspects, the PD-l axis binding antagonist is a PD-l binding antagonist. In certain aspects, the PD-l binding antagonist inhibits the binding of PD-l to PDL1 and/or PDL2. In particular, the PD- 1 binding antagonist is a monoclonal antibody or antigen binding fragment thereof. In some embodiments, the PD-l binding antagonist is nivolumab, pembrolizumab, pidilizumab, AMP- 514, REGN2810, CT-011, BMS 936559, MPDL3280A or AMP-224.

[0010] In some aspects, the at least one additional therapy is a histone deacetylase (HD AC) inhibitor. In certain aspects, the HD AC inhibitor is tractinostat (CHR-3996 or VRx- 3996). In certain aspects, the method further comprises providing an extracellular matrix degrading protein, such as relaxin, hyaluronidase or decorin.

[0011] In some aspects, the at least one additional anticancer treatment is an oncolytic vims. In some aspects, the oncolytic vims is engineered to express p53, MDA-7, IL-12, TGF- b inhibitor, and/or IL-10 inhibitor. In certain aspects, the oncolytic virus is a single- or double- stranded DNA vims, RNA vims, adenovirus, adeno-associated vims, retrovirus, lentivirus, herpes virus, pox virus, vaccinia virus, vesicular stomatitis virus, polio virus, Newcastle’s Disease vims, Epstein-Barr virus, influenza virus, reoviruses, myxoma virus, maraba virus, rhabdovims, enadenotucirev or coxsackie virus. In some aspects, the oncolytic virus is engineered to express a cytokine, such as granulocyte-macrophage colony- stimulating factor (GM-CSF) or IL-12. In some aspects, the oncolytic virus is further defined as talimogene laherparepvec (T-VEC). In some aspects, the oncolytic adenoviral vector is derived from an Elb deleted adenovirus, and adenovirus where the Ad Ela gene is driven by the alpha- fetoprotein (AFP) promoter, a modified TERT Promoter Oncolytic Adenovirus, the HRE-E2F- TERT Hybrid Promoter Oncolytic Adenovirus, and/or an adenovirus with a modified Ela regulatory sequence wherein at least one Pea3 binding site, or a functional portion thereof, is deleted with an Elb-l9K clone insertion site, which may all be modified to express therapeutic genes.

[0012] In certain aspects, the at least one additional anticancer treatment is a protein kinase or growth factor signaling pathways inhibitor. In certain aspects, the protein kinase or growth factor signaling pathways inhibitor is Afatinib, Axitinib, Bevacizumab, Bosutinib, Cetuximab, Crizotinib, Dasatinib, Erlotinib, Fostamatinib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Mubritinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Saracatinib, Sorafenib, Sunitinib, Trastuzumab, Vandetanib, AP23451, Vemurafenib, CAL101, PX-866, LY294002, rapamycin, temsirolimus, everolimus, ridaforolimus, Alvocidib, Genistein, Selumetinib, AZD-6244, Vatalanib, P1446A-05, AG- 024322, ZD1839, P276-00 or GW572016. In some aspects, the protein kinase inhibitor is a PI3K inhibitor, such as a PI3K delta inhibitor.

[0013] In some aspects, the immunotherapy comprises a cytokine, such as GM-CSF, an interleukin (e.g., IL-2) and/or an interferon (e.g., IFNa) or heat shock proteins. In certain aspects, the immunotherapy comprises a co-stimulatory receptor agonist, a stimulator of innate immune cells, or an activator of innate immunity. In certain aspects, the co- stimulatory receptor agonist is an anti-OX40 antibody, anti-GITR antibody, anti-CD 137 antibody, anti-CD40 antibody, or an anti-CD27 antibody. In some aspects, the stimulator of immune cells is an inhibitor of a cytotoxicity-inhibiting receptor or an agonist of immune stimulating toll like receptors (TLR). In some aspects, the cytotoxicity-inhibiting receptor is an inhibitor of NKG2A/CD94 or CD96 TACTILE. In some aspects, the TLR agonist is a TLR7 agonist, TLR8 agonist, or TLR9 agonist. In some aspects, the immunotherapy comprises a combination of a PD-L1 inhibitor, a 4-1BB agonist, and an 0X40 agonist. In certain aspects, the immunotherapy comprises a stimulator of interferon genes (STING) agonist. In some aspects, the activator of innate immunity is an IDO inhibitor, TGF inhibitor, or IL-10 inhibitor. In some aspects, when these immunotherapies are proteins, they may be delivered as polypeptides or their corresponding nucleic acids administered by replication competent and/or replication incompetent viral and/or non-viral gene therapy. In some aspects, the chemotherapy comprises a DNA damaging agent, such as gamma- irradiation, X-rays, UV-irradiation, microwaves, electronic emissions, adriamycin, 5- fluorouracil (5FU), capecitabine, etoposide (VP-16), camptothecin, actinomycin-D, mitomycin C, cisplatin (CDDP), or hydrogen peroxide.

[0014] In some aspects, the at least one additional anticancer therapy is at least one oncolytic vims. In some aspects, the at least one oncolytic vims is selected from the group consisting of a single- or double- stranded DNA virus, RNA vims, adenovirus, adeno- associated vims, retrovims, lentivims, herpes vims, pox virus, vaccinia vims, vesicular stomatitis virus, polio virus, Newcastle’s Disease vims, Epstein-Barr virus, influenza vims, reoviruses, myxoma virus, maraba vims, rhabdovims, enadenotucirev, and coxsackie virus.

[0015] In some aspects, the vimses employed in the above embodiments comprise replication competent and/or replication defective vimses. In certain aspects, the replication competent or replication incompetent vims is a single or double stranded DNA vims, RNA virus, adenovims, adeno-associated virus, retrovirus, lentivirus, herpes virus, pox virus, vaccinia vims, vesicular stomatitis virus, polio virus, Newcastle’s Disease vims, myxoma virus, Epstein-Barr vims, influenza virus, reovirus, maraba virus, rhabdovims, enadenotucirev or coxsackie virus. In certain aspects, one or more vimses are utilized. In certain aspects, the vims composition comprises a combination of replication competent and replication incompetent viruses.

[0016] In further aspects, the replication competent vimses in the above embodiments may be one or more oncolytic vimses. These oncolytic viruses may be engineered to express p53 and/or IL24 and/or to express a gene other than p53 and/or IL24, such as a cytokine (e.g. IL12) and/or another immune stimulatory gene (e.g., TGF-beta inhibitors or IL10 inhibitors or heat shock proteins). In certain aspects, the oncolytic virus may be used in lieu of or in addition to p53 and/or IL24 tumor suppressor therapy. Examples of oncolytic vimses include single or double stranded DNA vimses, RNA viruses, adenoviruses, adeno-associated vimses, retrovimses, lentiviruses, herpes vimses, pox viruses, vaccinia viruses, vesicular stomatitis vimses, polio vimses, Newcastle’s Disease viruses, Epstein-Barr vimses, influenza vimses and reoviruses, myxoma vimses, maraba vimses, rhabdovimses, enadenotucirev or coxsackie vimses. Exemplary oncolytic viruses include, but are not limited to, Ad5- y CD/mutTKS R39rep -hIL 12 , Cavatak™, CG0070, DNX-2401, G207, HF10, IMLYGIC™, JX-594, MG1-MA3, MV-NIS, OBP-301, Reolysin®, Toca 511, Oncorine (H101), Onyx-Ol5, H102, H103, RIGVIR, an adenovirus overexpressing the adenoviral death protein (ADP), such as VirRx007, an N1L deleted vaccinia virus or an N1L deleted vaccinia virus expressing IL12.

[0017] In certain aspects of the above embodiments, the method further comprises providing an extracellular matrix-degrading protein. In some aspects, this comprises administering an expression cassette encoding the extracellular matrix-degrading protein. In some embodiments, the extracellular matrix-degrading protein is relaxin, hyaluronidase or decorin. In particular aspects, the extracellular matrix-degrading protein is relaxin. In some aspects, the expression cassette is in a viral vector. In certain aspects, the viral vector is an adenoviral vector, a retroviral vector, a vaccinia viral vector, an adeno-associated viral vector, a herpes viral vector, a vesicular stomatitis viral vector, or a polyoma viral vector or another type of viral or non- viral gene therapy vector.

[0018] In some aspects, the subject is further administered a tumor suppressor immune gene therapy. In some aspects, the subject is further administered additional viral and/or non- viral gene therapies.

[0019] In particular aspects, the treated subject is a mammal or human. In certain aspects, the treatment is provided to prevent or treat a pre-malignant or a malignant hyperproliferative condition. In certain aspects of prevention, the subject is a healthy subject. In other aspects of prevention, the subject comprises a pre-malignant lesion, such as, for example, a leukoplakia or a dysplastic lesion. In other aspects of prevention, the subject is at risk of developing cancer, such as, for example, by being a smoker or having a family history of cancer. In certain aspects, the treatment is for initial or recurrent hyperproliferative conditions. In some aspects, the treatment is administered to augment or reverse resistance to another therapy. In certain aspects, the resistance to treatment is known historically for a particular population of hyperproliferative condition patients. In certain aspects, the resistance to treatment is observed in individual hyperproliferative condition patients.

[0020] In some aspects, the p53 dendritic cell vaccine composition is administered to non-tumor sites by routes utilized for infectious disease vaccines which include intradermally, subcutaneously, intramuscularly, intra-peritoneally, orally, by inhalation, or by other forms of mucosal exposure. In other aspects, the p53 dendritic cell vaccine composition is administered to tumor sites intratumorally, intraarterially, intravenously, intravascularly, intrapleuraly, intraperitoneally, intratracheally, intrathecally, intramuscularly, endoscopically, intralesionally, percutaneously, subcutaneously, regionally, stereotactically, or by direct injection or perfusion. In some aspects, administering is via continuous infusion, intratumoral injection, intravenous injection, intra-arterial injection, intra-peritoneal injection, intrapleural injection, or intra-thecal injection. In certain aspects, the p53 dendritic cell vaccine composition is administered to non-tumor sites and to tumor sites either sequentially or concurrently.

[0021] In certain aspects, the subject is administered the p53 dendritic cell vaccine composition after the at least one preferential CD122/CD132 agonist. In certain aspects, the subject is administered the p53 dendritic cell vaccine composition before the at least one preferential CD122/CD132 agonist. In certain aspects, the subject is administered the p53 dendritic cell vaccine composition simultaneously with the at least one preferential CD122/CD132 agonist. In some aspects, the p53 dendritic cell vaccine composition is administered to the subject at non-tumor sites and enhances therapeutic effects or reverses resistance to CD122/CD132 agonist and/or immune checkpoint inhibitor treatments on distant tumors. In some aspects, the p53 dendritic cell vaccine composition is administered to the subject at tumor sites and enhances therapeutic effects or reverses resistance to CD122/CD132 agonist and/or immune checkpoint inhibitor treatments on distant tumors that are not injected with the p53 dendritic cell vaccine composition.

[0022] In some embodiments, the p53 dendritic cell vaccine component comprises administering at least 1, 2, 3, 4, 5 or 6 doses of dendritic cells per treatment cycle at 1 x 10⁶ to 1 x 10⁷ cells per dose, including about 1 x 10⁶, 2 x 10⁶, 3 x 10⁶, 4 x 10⁶, 5 x 10⁶, 6 x 10⁶, 7 x 10⁶, 8 x 10⁶, 9 x 10⁶, and 1 x 10⁷ cells per dose. These doses can be administered at 1, 2, 3, 4, 5, or 6 day intervals or 1, 2, 3, or 4 week intervals. Multiple cycles of p53 dendritic cell vaccinations can be given.

[0023] In certain embodiments, the p53 dendritic cell vaccine component comprises the sole vaccine for tumor antigen priming and booster immunizations. In some embodiments, the p53 dendritic cell vaccine component is combined with other p53 vaccines for either priming and/or boosting the anti-p53 immune response. In some aspects the other p53 vaccine may be a protein or peptide vaccine, an RNA or DNA vaccine, and/or a genetically engineered vims vaccine expressing p53. In certain aspects, the p53 priming and boosting vaccinations comprise the same or different p53 vaccine constructs. In particular aspects, the priming and boosting vaccinations are given by the same route of administration to non-tumor sites while in other embodiments the p53 vaccines are given by different routes of administration to non tumor sites. In some embodiments the p53 vaccines are given to non-tumor sites and to tumor sites. In certain aspects, the viral vaccines expressing p53 are replication competent or replication incompetent. The p53 viral vaccines may be a single or double stranded DNA vims, RNA vims, adenovims, adeno-associated vims, retrovims, lentivirus, herpes vims, pox virus, vaccinia vims, vesicular stomatitis virus, polio vims, Newcastle’s Disease vims, myxoma vims, Epstein-Barr vims, influenza virus, reovirus, maraba virus, rhabdovims, enadenotucirev or coxsackie vims. In certain aspects, one or more viruses are utilized.

[0024] In certain aspects, the cancer is melanoma, non-small cell lung, small-cell lung, lung, hepatocarcinoma, retinoblastoma, astrocytoma, glioblastoma, leukemia, neuroblastoma, head, neck, breast, pancreatic, prostate, renal, bone, testicular, ovarian, mesothelioma, cervical, gastrointestinal, urogenital, respiratory tract, hematopoietic, musculoskeletal, neuroendocrine, carcinoma, sarcoma, central nervous system, peripheral nervous system, lymphoma, brain, colon or bladder cancer. In some aspects, the cancer is metastatic.

[0025] In some aspects, the methods further comprise administering at least one additional anticancer treatment. In certain aspects, the at least one additional anticancer treatment is surgical therapy, chemotherapy (e.g., administration of a protein kinase inhibitor or a EGFR- targeted therapy), embolization therapy, chemoembolization therapy, radiation therapy, cryotherapy, hyperthermia treatment, phototherapy, radioablation therapy, hormonal therapy, immunotherapy, small molecule therapy, receptor kinase inhibitor therapy, anti- angiogenic therapy, cytokine therapy or a biological therapies such as monoclonal antibodies, siRNA, miRNA, antisense oligonucleotides, ribozymes or gene therapy. In particular aspects, the at least one additional anticancer treatment is a protein kinase inhibitor, such as a tyrosine kinase inhibitor. In one specific aspect, the protein kinase inhibitor is a Bruton’s tyrosine kinase (BTK) inhibitor (e.g., ibrutinib, acalabrutinib (ACP-196), ONO-4059, spebrutinib (CC-292), HM-71224, CG-036806, GDC-0834, ONO-4049, RN-486, SNS-062, TAS-5567, AVL-101, AVL-291, PCI-45261, HCI-1684, PLS-123, or BGB-3111). In some aspects, one or more BTK inhibitors are administered in combination with the virus composition. In certain aspects, one or more BTK inhibitors are administered in combination with the vims composition and at least one immune checkpoint inhibitor. In some aspects, the at least one additional anticancer treatment is an inhibitor (e.g., small molecule inhibitor) of HDM2 (also known as MDM2) and/or HDM4, such as to reverse its inhibition of p53 activity. In specific aspects, the small molecule inhibitor of HDM2 is HDM201, cis-imidazolines (e.g., Nutlins), benzodiazepines (BDPs), spiro-oxindoles.

[0026] In some aspects, the immunotherapy comprises a cytokine. In particular aspects, the cytokine is granulocyte macrophage colony-stimulating factor (GM-CSF), an interleukin such as IL-2, and/or an interferon such as IFN-alpha. Additional approaches to boost tumor-targeted immune responses include additional immune checkpoint inhibition. In some aspects, the immune checkpoint inhibition includes anti-CTLA4, anti-PD-l, anti-PD- Ll, anti-PD-L2, anti-TIM-3, anti-LAG-3, anti-A2aR, or anti-KIR antibodies. In some aspects, the immunotherapy comprises co-stimulatory receptor agonists such as anti-OX40 antibody, anti-GITR antibody, anti-CD 137 antibody, anti-CD40 antibody, and anti-CD27 antibody. In certain aspects, the immunotherapy comprises suppression of T regulatory cells (Tregs), myeloid derived suppressor cells (MDSCs) and cancer associated fibroblasts (CAFs). In further aspects, the immunotherapy comprises stimulation of innate immune cells, such as natural killer (NK) cells, macrophages, and dendritic cells. Additional immune stimulatory treatments may include IDO inhibitors, TGF-beta inhibitors, IL-10 inhibitors, stimulator of interferon genes (STING) agonists, toll like receptor (TLR) agonists (e.g., TLR7, TLR8, or TLR9), tumor vaccines (e.g., whole tumor cell vaccines, peptides, and recombinant tumor associated antigen vaccines), and adoptive cellular therapies(ACT) (e.g., T cells, natural killer cells, TILs, and LAK cells), and ACT with genetically engineered receptors (e.g., chimeric antigen receptors (CAR) and T cell receptors (TCR). In certain aspects, combinations of these agents may be used such as combining immune checkpoint inhibitors, checkpoint inhibition plus agonism of T-cell costimulatory receptors, and checkpoint inhibition plus TIL ACT. In certain aspects, additional anti-cancer treatment includes a combination of an immune checkpoint inhibitor (e.g., Avelumab), a 4-1BB (CD-137) agonist (e.g. Utomilumab), and an 0X40 (TNFRS4) agonist.

[0027] In some aspects, the chemotherapy comprises a DNA damaging agent. In some embodiments, the DNA damaging agent is gamma- irradiation, X-rays, UV-irradiation, microwaves, electronic emissions, adriamycin, 5- fluorouracil (5FU), capecitabine, etoposide (VP-16). camptothecin. actinomycin-D, mitomycin C, cisplatin (CDDP), or hydrogen peroxide. In particular aspects, the DNA damaging agent is 5FU or capecitabine. In some aspects, the chemotherapy comprises a cisplatin (CDDP), carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, bisulfan, nitrosurea, dactinomycin, daunorubicin, doxombicin, bleomycin, plicomycin, mitomycin, etoposide (VP16), tamoxifen, taxotere, taxol, transplatinum, 5-fluorouracil, vincristin, vinblastin, methotrexate, an HDAC inhibitor or any analog or derivative variant thereof.

[0028] In some aspects, the at least one additional anticancer treatment is a replication competent or replication incompetent vims. In certain aspects, the replication competent or replication incompetent virus is a single or double stranded DNA virus, RNA virus, adenovirus, adeno-associated vims, retrovims, lentivims, herpes virus, pox vims, vaccinia vims, vesicular stomatitis virus, polio virus, Newcastle’s Disease vims, myxoma vims, Epstein-Barr vims, influenza vims, reovims, maraba vims, rhabdovims, enadenotucirev or coxsackie virus. In particular aspects, the replication competent or replication incompetent vims is herpes simplex vims. In some aspects, the replication competent or replication incompetent vims is engineered to express a transgene, such as a tumor suppressor (e.g., p53) and/or a cytokine (e.g., IL-24 or IL-12) and/or heat shock proteins. In some embodiments, the cytokine is granulocyte- macrophage colony- stimulating factor (GM-CSF). In some embodiments, the replication competent or replication incompetent virus is further defined as talimogene laherparepvec (T- VEC) (e.g., IMLYGIC™). In some embodiments, the additional replication competent or replication incompetent virus is administered before, simultaneously, or after the p53 dendritic cell therapeutic vaccine and CD122/CD132 agonist therapy.

[0029] In some aspects, the at least one additional cancer treatment is a protein kinase inhibitor or a monoclonal antibody that inhibits receptors involved in protein kinase or growth factor signaling pathways. For example, the protein kinase or receptor inhibitor can be an EGER, VEGFR, AKT, Erbl, Erb2, ErbB, Syk, Bcr-Abl, JAK, Src, GSK-3, PI3K, Ras, Raf, MAPK, MAPKK, mTOR, c-Kit, eph receptor or BRAF inhibitor. In particular aspects, the protein kinase inhibitor is a PI3K inhibitor. In some embodiments, the PI3K inhibitor is a PI3K delta inhibitor. For example, the protein kinase or receptor inhibitor can be Afatinib, Axitinib, Bevacizumab, Bosutinib, Cetuximab, Crizotinib, Dasatinib, Erlotinib, Fostamatinib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Mubritinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Saracatinib, Sorafenib, Sunitinib, Trastuzumab, Vandetanib, AP23451, Vemurafenib, CAL101, PX-866, LY294002, rapamycin, temsirolimus, everolimus, ridaforolimus, Alvocidib, Genistein, Selumetinib, AZD-6244, Vatalanib, P1446A-05, AG- 024322, ZD1839, P276-00, GW572016, or a mixture thereof. In certain aspects, the protein kinase inhibitor is an AKT inhibitor (e.g., MK-2206, GSK690693, A-443654, VQD-002, Miltefosine or Perifosine). In certain aspects, EGFR-targeted therapies for use in accordance with the embodiments include, but are not limited to, inhibitors of EGFR/ErbBl/HER, ErbB2/Neu/HER2, ErbB3/HER3, and/or ErbB4/HER4. A wide range of such inhibitors are known and include, without limitation, tyrosine kinase inhibitors active against the receptor(s) and EGFR-binding antibodies or aptamers. For instance, the EGFR inhibitor can be gefitinib, erlotinib, cetuximab, matuzumab, panitumumab, AEE788; 0-1033, HKI-272, HKI-357, or EKB-569. The protein kinase inhibitor may be a BRAF inhibitor such as dabrafenib, or a MEK inhibitor such as trametinib.

[0030] In a further embodiment, there is provided a pharmaceutical composition comprising (a) p53 dendritic cell therapeutic vaccine (b) at least one preferential CD122/CD132 agonist therapy; (c) at least one immune checkpoint inhibitor and (d) at least one HDAC inhibitor. In certain aspects, the p53 dendritic cell therapeutic vaccine is generated by infecting dendritic cells with an adenovirus expressing a p53 gene.

[0031] Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0033] FIG. 1: Generation and Administration of p53 Dendritic Cells. The patient has a leukapheresis from which peripheral blood mononuclear cells (PBMC) are isolated by Ficoll separation. The PBMCs are cultured ex vivo with GM-CSF and IL-4 to produce autologous dendritic cells (DC). A replication defective adenoviral vector expressing the wild type p53 gene (Ad-p53) is utilized ex vivo to transduce the patient’s DCs which are employed as antigen presenting cells for the final product which is then used to vaccinate the patient.

[0034] FIGS. 2A-2C: Tumor Responses. It is generally appreciated that tumor responses with reductions in tumor size following treatment are associated with important therapeutic benefits. FIG. 2A depicts the tumor response rates of reductions in tumor size > 50% for each of the treatment groups. There was an unexpectedly high rate of tumor responses in the Ad-p53 DC vaccine + CD 122/132 + anti-PD-l treatment group with 87.5% (7/8 animals) demonstrating a > 50% reduction in tumor volume. None of the other treatment combinations demonstrated synergistic effects as their combined response rates were either similar or lower to each of their monotherapy results. For example, the Ad-p53 DC vaccine and CD122/CD132 treatment combination appeared to have an inhibitory effect with 0% responders. The combination of CD122/CD132 and anti-PD-l treatment had no greater response rate than CD122/CD132 monotherapy. A Chi-square analysis comparing the treatment groups revealed a highly statistically significant difference between the treatments (p=0.0044). Fisher’s exact test demonstrated a statistically significant increased response rate for tumor size reductions > 50% in the Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group (p-value = 0.0010 by two-sided Fisher’s Exact Test comparing Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group vs. animals in all other Ad-p53 DC vaccine, CD 122/132 and anti-PD-l treatment groups). The specific tumor size reductions in the Ad-p53 DC vaccine + CD 122/132 + anti-PD-l treatment group are shown in FIG. 2B. The overall mean tumor volume reductions for the treatment groups are shown in FIG. 2C. There was a surprisingly high mean tumor volume reduction in the Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group (- 67.1%). A statistical analysis of variance (ANOVA) comparison of these mean tumor volume reductions was statistically significant (p-value = 0.0013). Only the Ad-p53 DC vaccine + CD122/132 + anti-PD-l group demonstrated a statistically significant decrease in mean tumor volume reduction vs. the vehicle control group (p-value = 0.0005). None of the other treatment combinations demonstrated synergistic or even substantial additive effects as their mean tumor volume reductions were either similar or lower to each of their monotherapy results. The synergistic mean tumor volume reduction for the Ad-p53 DC vaccine + CD 122/132 + anti-PD- 1 treatment group was highly remarkable because the combination of Ad-p53 DC + CD122/132 treatment had a significant inhibitory effect on efficacy. DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0035] It is well known that tumors evolve during their initiation and progression to evade destruction by the immune system. While the recent use of immune checkpoint inhibitors and other immune therapies to reverse this resistance has demonstrated some success, the majority of patients do not respond these treatments. In certain embodiments, the present disclosure provides methods and compositions for inducing anti-p53 immune responses for the treatment and prevention of cancer.

[0036] In certain embodiments, the present disclosure provides methods to treat cancer by administering an effective amount of a dendritic cell therapeutic vaccine incorporating the p53 tumor suppressor as an antigen to generate an anti-p53 immune response and administering at least one preferential CD122/CD132 agonist to treat a cancer in a subject. In this p53 dendritic cell therapeutic vaccine CD122/CD132 agonist therapy, the CD122/CD132 agonist in the present disclosure may preferentially bind to the CD 122/CD 132 receptor complex and have lower affinity binding for CD25 or the ILl5a receptor. In certain aspects, more than one preferential CD122/CD132 agonist is administered.

[0037] It is generally appreciated that tumor responses with reductions in tumor size following treatment are associated with important therapeutic benefits. There was an unexpected, surprisingly high rate of tumor responses in the Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group with 87.5% demonstrating a > 50% reduction in tumor volume. None of the other treatment combinations demonstrated synergistic or substantial additive effects as their combined response rates were either similar or lower to each of their monotherapy results. There was a surprisingly high mean tumor volume reduction in the Ad- p53 DC vaccine + CD122/132 + anti-PD-l treatment group (-67.1%). None of the other treatment combinations demonstrated synergistic or substantial additive effects as their mean tumor volume reductions were either similar or lower to each of their monotherapy results. The synergistic mean tumor volume reduction Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group for the Ad-p53 DC and CD122/CD132 treatment combination was remarkable because the combination of Ad-p53 DC + CD122/132 treatment had a significant inhibitory effect on efficacy.

[0038] The preferential CD122/CD132 agonist may be an IL-2/anti-IL-2 immune complex, an IL-l5/anti-IL-l5 immune complex, an IL-15/IL-15 Receptor a-IgGl-Fc (IL- l5/IL-l5Ra-IgGl-Fc) immune complex, PEGylated IL-2, PEGylated IL-15, IL-2 mutein and/or IL-15 mutein or PEGylated IL-2 mutein and/or IL-15 mutein. Additional types of CD122/132 agonists are listed in Table 1.

[0039] The p53 dendritic cell therapeutic vaccine composition may comprise dendritic cells engineered to present p53 tumor antigen epitopes by transduction with replication incompetent adenoviral p53 vectors in combination with a preferential CD 122/CD 132 agonist. Particularly, the Ad-p53 dendritic cell therapeutic vaccine composition may be administered to a non-tumor site intra-dermally, intra-muscularly or subcutaneously while the preferential CD 122/CD 132 agonist is administered intravenously or subcutaneously.

[0040] In some embodiments, the p53 dendritic cell therapeutic vaccine and CD 122/CD 132 agonist treatment is administered in combination with one or more immune checkpoint inhibitors such as an anti-PDl antibody or an anti-CTLA4 antibody to enhance the immune response. The p53 dendritic cell therapeutic vaccine CD122/CD132 agonist treatment may be administered before, concurrently with, or after immune checkpoint inhibitor therapy.

[0041] Further, the methods of treatment can include additional anti-cancer therapies such as HDAC inhibitors, oncolytic viruses, replication incompetent viral gene therapy, cytokines or chemotherapeutics to enhance the anti-tumor effect of the combination therapy provided herein. For example, the oncolytic virus could be a replication competent adenovirus or vaccinia virus, the cytokine could be GM-CSF and the chemotherapy could be 5-fluorouracil (5FU), capecitabine, cyclophosphamide, or a PI3K inhibitor. The replication competent and/or replication incompetent viral gene therapy may deliver one or more therapeutic genes which could be tumor suppressor genes or immune stimulatory genes.

I. Definitions

[0042] As used herein,“essentially free,” in terms of a specified component, is used herein to mean that none of the specified component has been purposefully formulated into a composition and/or is present only as a contaminant or in trace amounts. The total amount of the specified component resulting from any unintended contamination of a composition is therefore well below 0.05%, preferably below 0.01%. Most preferred is a composition in which no amount of the specified component can be detected with standard analytical methods. [0043] As used herein the specification,“a” or“an” may mean one or more. As used herein in the claim(s), when used in conjunction with the word“comprising,” the words“a” or “an” may mean one or more than one.

[0044] The use of the term“or” in the claims is used to mean“and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and“and/or.” As used herein “another” may mean at least a second or more.

[0045] Throughout this application, the term“about” is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value, or the variation that exists among the study subjects.

[0046] As used herein“wild-type” refers to the naturally occurring sequence of a nucleic acid at a genetic locus in the genome of an organism, and sequences transcribed or translated from such a nucleic acid. Thus, the term "wild-type" also may refer to the amino acid sequence encoded by the nucleic acid. As a genetic locus may have more than one sequence or alleles in a population of individuals, the term“wild-type” encompasses all such naturally occurring alleles. As used herein the term“polymorphic” means that variation exists (/._<?., two or more alleles exist) at a genetic locus in the individuals of a population. As used herein, "mutant" refers to a change in the sequence of a nucleic acid or its encoded protein, polypeptide, or peptide that is the result of recombinant DNA technology.

[0047] The term“exogenous,” when used in relation to a protein, gene, nucleic acid, or polynucleotide in a cell or organism refers to a protein, gene, nucleic acid, or polynucleotide that has been introduced into the cell or organism by artificial or natural means; or in relation to a cell, the term refers to a cell that was isolated and subsequently introduced to other cells or to an organism by artificial or natural means. An exogenous nucleic acid may be from a different organism or cell, or it may be one or more additional copies of a nucleic acid that occurs naturally within the organism or cell. An exogenous cell may be from a different organism, or it may be from the same organism. By way of a non-limiting example, an exogenous nucleic acid is one that is in a chromosomal location different from where it would be in natural cells, or is otherwise flanked by a different nucleic acid sequence than that found in nature. [0048] By“expression construct” or“expression cassette” is meant a nucleic acid molecule that is capable of directing transcription. An expression construct includes, at a minimum, one or more transcriptional control elements (such as promoters, enhancers or a structure functionally equivalent thereof) that direct gene expression in one or more desired cell types, tissues or organs. Additional elements, such as a transcription termination signal, may also be included.

[0049] A“vector” or“construct” (sometimes referred to as a gene delivery system or gene transfer“vehicle”) refers to a macromolecule or complex of molecules comprising a polynucleotide to be delivered to a host cell, either in vitro or in vivo.

[0050] A“plasmid,” a common type of a vector, is an extra-chromosomal DNA molecule separate from the chromosomal DNA that is capable of replicating independently of the chromosomal DNA. In certain cases, it is circular and double- stranded.

[0051] An“origin of replication” (“ori”) or“replication origin” is a DNA sequence, e.g., in a lymphotrophic herpes virus, that when present in a plasmid in a cell is capable of maintaining linked sequences in the plasmid and/or a site at or near where DNA synthesis initiates. As an example, an ori for EBV includes FR sequences (20 imperfect copies of a 30 bp repeat), and preferably DS sequences; however, other sites in EBV bind EBNA-l, e.g., Rep* sequences can substitute for DS as an origin of replication (Kirshmaier and Sugden, 1998). Thus, a replication origin of EBV includes FR, DS or Rep* sequences or any functionally equivalent sequences through nucleic acid modifications or synthetic combination derived therefrom. For example, the present invention may also use genetically engineered replication origin of EBV, such as by insertion or mutation of individual elements, as specifically described in Lindner, et. al, 2008.

[0052] A “gene,” “polynucleotide,” “coding region,” “sequence,” “segment,” “fragment,” or“transgene” that“encodes” a particular protein, is a nucleic acid molecule that is transcribed and optionally also translated into a gene product, e.g., a polypeptide, in vitro or in vivo when placed under the control of appropriate regulatory sequences. The coding region may be present in either a cDNA, genomic DNA, or RNA form. When present in a DNA form, the nucleic acid molecule may be single-stranded (/._<?., the sense strand) or double-stranded. The boundaries of a coding region are determined by a start codon at the 5' (amino) terminus and a translation stop codon at the 3' (carboxy) terminus. A gene can include, but is not limited to, cDNA from prokaryotic or eukaryotic mRNA, genomic DNA sequences from prokaryotic or eukaryotic DNA, and synthetic DNA sequences. A transcription termination sequence will usually be located 3' to the gene sequence.

[0053] The term “control elements” refers collectively to promoter regions, polyadenylation signals, transcription termination sequences, upstream regulatory domains, origins of replication, internal ribosome entry sites (IRES), enhancers, splice junctions, and the like, which collectively provide for the replication, transcription, post-transcriptional processing, and translation of a coding sequence in a recipient cell. Not all of these control elements need be present so long as the selected coding sequence is capable of being replicated, transcribed, and translated in an appropriate host cell.

[0054] The term“promoter” is used herein in its ordinary sense to refer to a nucleotide region comprising a DNA regulatory sequence, wherein the regulatory sequence is derived from a gene that is capable of binding RNA polymerase and initiating transcription of a downstream (3' direction) coding sequence. It may contain genetic elements at which regulatory proteins and molecules may bind, such as RNA polymerase and other transcription factors, to initiate the specific transcription of a nucleic acid sequence. The phrases “operatively positioned,”“operatively linked,”“under control,” and“under transcriptional control” mean that a promoter is in a correct functional location and/or orientation in relation to a nucleic acid sequence to control transcriptional initiation and/or expression of that sequence.

[0055] By “enhancer” is meant a nucleic acid sequence that, when positioned proximate to a promoter, confers increased transcription activity relative to the transcription activity resulting from the promoter in the absence of the enhancer domain.

[0056] By“operably linked” or co-expressed” with reference to nucleic acid molecules is meant that two or more nucleic acid molecules (e.g., a nucleic acid molecule to be transcribed, a promoter, and an enhancer element) are connected in such a way as to permit transcription of the nucleic acid molecule.“Operably linked” or“co-expressed” with reference to peptide and/or polypeptide molecules means that two or more peptide and/or polypeptide molecules are connected in such a way as to yield a single polypeptide chain, /._<?., a fusion polypeptide, having at least one property of each peptide and/or polypeptide component of the fusion. The fusion polypeptide is preferably chimeric, i.e., composed of heterologous molecules.

[0057]“Homology” refers to the percent of identity between two polynucleotides or two polypeptides. The correspondence between one sequence and another can be determined by techniques known in the art. For example, homology can be determined by a direct comparison of the sequence information between two polypeptide molecules by aligning the sequence information and using readily available computer programs. Alternatively, homology can be determined by hybridization of polynucleotides under conditions that promote the formation of stable duplexes between homologous regions, followed by digestion with single strand-specific nuclease(s), and size determination of the digested fragments. Two DNA, or two polypeptide, sequences are“substantially homologous” to each other when at least about 80%, preferably at least about 90%, and most preferably at least about 95% of the nucleotides, or amino acids, respectively match over a defined length of the molecules, as determined using the methods above.

[0058] The term“nucleic acid" will generally refer to at least one molecule or strand of DNA, RNA or a derivative or mimic thereof, comprising at least one nucleobase, such as, for example, a naturally occurring purine or pyrimidine base found in DNA (e.g., adenine“A,” guanine“G,” thymine“T,” and cytosine“C”) or RNA (e.g. A, G, uracil“U,” and C). The term "nucleic acid" encompasses the terms“oligonucleotide” and“polynucleotide.” The term “oligonucleotide” refers to at least one molecule of between about 3 and about 100 nucleobases in length. The term“polynucleotide” refers to at least one molecule of greater than about 100 nucleobases in length. These definitions generally refer to at least one single- stranded molecule, but in specific embodiments will also encompass at least one additional strand that is partially, substantially or fully complementary to the at least one single- stranded molecule. Thus, a nucleic acid may encompass at least one double-stranded molecule or at least one triple- stranded molecule that comprises one or more complementary strand(s) or“complement(s)” of a particular sequence comprising a strand of the molecule.

[0059] The term“therapeutic benefit” used throughout this application refers to anything that promotes or enhances the well-being of the patient with respect to the medical treatment of his cancer. A list of nonexhaustive examples of this includes extension of the patient's life by any period of time; decrease or delay in the neoplastic development of the disease; decrease in hyperproliferation; reduction in tumor growth; delay of metastases; reduction in the proliferation rate of a cancer cell or tumor cell; induction of apoptosis in any treated cell or in any cell affected by a treated cell; and a decrease in pain to the patient that can be attributed to the patient's condition.

[0060] An“effective amount” is at least the minimum amount required to effect a measurable improvement or prevention of a particular disorder. An effective amount herein may vary according to factors such as the disease state, age, sex, and weight of the patient, and the ability of the antibody to elicit a desired response in the individual. An effective amount is also one in which any toxic or detrimental effects of the treatment are outweighed by the therapeutically beneficial effects. For prophylactic use, beneficial or desired results include results such as eliminating or reducing the risk, lessening the severity, or delaying the onset of the disease, including biochemical, histological and/or behavioral symptoms of the disease, its complications and intermediate pathological phenotypes presenting during development of the disease. For therapeutic use, beneficial or desired results include clinical results such as decreasing one or more symptoms resulting from the disease, increasing the quality of life of those suffering from the disease, decreasing the dose of other medications required to treat the disease, enhancing effect of another medication such as via targeting, delaying the progression of the disease, and/or prolonging survival. In the case of cancer or tumor, an effective amount of the drug may have the effect in reducing the number of cancer cells; reducing the tumor size; inhibiting (/._<?., slow to some extent or desirably stop) cancer cell infiltration into peripheral organs; inhibit (/._<?., slow to some extent and desirably stop) tumor metastasis; inhibiting to some extent tumor growth; and/or relieving to some extent one or more of the symptoms associated with the disorder. An effective amount can be administered in one or more administrations. For purposes of this invention, an effective amount of drug, compound, or pharmaceutical composition is an amount sufficient to accomplish prophylactic or therapeutic treatment either directly or indirectly. As is understood in the clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable result may be or is achieved.

[0061] As used herein,“carrier” includes any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic and absorption delaying agents, buffers, carrier solutions, suspensions, colloids, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.

[0062] The term“pharmaceutical formulation” refers to a preparation which is in such form as to permit the biological activity of the active ingredient to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered. Such formulations are sterile. “Pharmaceutically acceptable” excipients (vehicles, additives) are those which can reasonably be administered to a subject mammal to provide an effective dose of the active ingredient employed.

[0063] As used herein, the term“treatment” refers to clinical intervention designed to alter the natural course of the individual or cell being treated during the course of clinical pathology. Desirable effects of treatment include decreasing the rate of disease progression, ameliorating or palliating the disease state, and remission or improved prognosis. For example, an individual is successfully“treated” if one or more symptoms associated with cancer are mitigated or eliminated, including, but are not limited to, reducing the proliferation of (or destroying) cancerous cells, decreasing symptoms resulting from the disease, increasing the quality of life of those suffering from the disease, decreasing the dose of other medications required to treat the disease, and/or prolonging survival of individuals.

[0064] An“anti-cancer” agent is capable of negatively affecting a cancer cell/tumor in a subject, for example, by promoting killing of cancer cells, inducing apoptosis in cancer cells, reducing the growth rate of cancer cells, reducing the incidence or number of metastases, reducing tumor size, inhibiting tumor growth, reducing the blood supply to a tumor or cancer cells, promoting an immune response against cancer cells or a tumor, preventing or inhibiting the progression of cancer, or increasing the lifespan of a subject with cancer.

[0065] The term“antibody” herein is used in the broadest sense and specifically covers monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g. , bispecific antibodies), and antibody fragments so long as they exhibit the desired biological activity. [0066] The term“monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible mutations, e.g., naturally occurring mutations, that may be present in minor amounts. Thus, the modifier“monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies. In certain embodiments, such a monoclonal antibody typically includes an antibody comprising a polypeptide sequence that binds a target, wherein the target-binding polypeptide sequence was obtained by a process that includes the selection of a single target binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, or recombinant DNA clones. It should be understood that a selected target binding sequence can be further altered, for example, to improve affinity for the target, to humanize the target binding sequence, to improve its production in cell culture, to reduce its immunogenicity in vivo, to create a multispecific antibody, etc. , and that an antibody comprising the altered target binding sequence is also a monoclonal antibody of this invention. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.

[0067] The term“p53 dendritic cell vaccine” refers to dendritic cells which present p53 epitopes for antigen presentation to generate anti-p53 immune responses. The p53 dendritic cell vaccine may be generated by combining dendritic cells with p53 polypeptides, or by transducing dendritic cells or antigen presenting cells with adenoviruses expressing a p53 transgene.

[0068] The term“CD122/CD132 agonist” or“preferential CD122/CD132 agonist” refers to an agent that preferentially binds to the CD 122/CD 132 receptor complex and has lower affinity binding for the IL-2 a receptor (CD25) or the IL-15 a receptor. Known preferential CD 122/CD 132 agonists comprise an IL2/anti-IL2 monoclonal antibody immunocomplex (see, for example, U.S. Patent Publication No. US20170183403A1; incorporated herein by reference in its entirety); a genetically engineered IL-2 mutein that has a modified amino acid sequence compared to wild type IL-2 (see, for example, U.S. Patent Publication No. US 2017/0044229 Al ; incorporated herein by reference in its entirety); a genetically engineered IL-2 mutein that has a modified amino acid sequence compared to wild type IL-2 combined with an anti-IL2 monoclonal antibody immunocomplex (see, for example, International Patent Publication No. W02014100014A1; incorporated herein by reference in its entirety); a PEGylated form of IL-2, such as NKTR-214 (see, for example, Charych et al, 2016; incorporated herein by reference in its entirety), an IL-l5/anti-IL-l5 monoclonal antibody immunocomplex; an IL15/IL15 Receptor a-IgGl-Fc (ILl5/ILl5Ra-IgGl-Fc) immunocomplex (see, for example, U.S. Patent Publication No. US20060257361A1, EP2724728A1 and Dubois et al., 2008; all incorporated herein by reference); a genetically engineered IL-15 mutein that has a modified amino acid sequence compared to wild type IL- 15 combined with an ILl5Ra-IgGl-Fc immunocomplex (see, for example, U.S. Patent Publicaiton No. US20070160578; incorporated herein by reference in its entirety); or a PEGylated form of IL-15 with preferential binding to CD122/CD132.

[0069] The term“immune checkpoint” refers to a molecule such as a protein in the immune system which provides inhibitory signals to its components in order to balance immune reactions. Known immune checkpoint proteins comprise CTLA-4, PD-l and its ligands PD-L1 and PD-L2 and in addition LAG-3, BTLA, B7H3, B7H4, TIM3, KIR. The pathways involving LAG3, BTLA, B7H3, B7H4, TIM3, and KIR are recognized in the art to constitute immune checkpoint pathways similar to the CTLA-4 and PD-l dependent pathways (see e.g. Pardoll, 2012. Nature Rev Cancer 12:252-264; Mellman et al, 2011. Nature 480:480- 489).

[0070] The term“PD-l axis binding antagonist” refers to a molecule that inhibits the interaction of a PD-l axis binding partner with either one or more of its binding partners, so as to remove T-cell dysfunction resulting from signaling on the PD-l signaling axis - with a result being to restore or enhance T-cell function (e.g., proliferation, cytokine production, target cell killing). As used herein, a PD-l axis binding antagonist includes a PD-l binding antagonist, a PD-L1 binding antagonist and a PD-L2 binding antagonist.

[0071] The term“PD-l binding antagonist” refers to a molecule that decreases, blocks, inhibits, abrogates or interferes with signal transduction resulting from the interaction of PD-l with one or more of its binding partners, such as PD-L1 and/or PD-L2. In some embodiments, the PD- 1 binding antagonist is a molecule that inhibits the binding of PD- 1 to one or more of its binding partners. In a specific aspect, the PD-l binding antagonist inhibits the binding of PD-l to PD-L1 and/or PD-L2. For example, PD-l binding antagonists include anti-PD-l antibodies, antigen binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that decrease, block, inhibit, abrogate or interfere with signal transduction resulting from the interaction of PD-l with PD-L1 and/or PD-L2. In one embodiment, a PD-l binding antagonist reduces the negative co- stimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes mediated signaling through PD-l so as render a dysfunctional T-cell less dysfunctional (e.g., enhancing effector responses to antigen recognition). In some embodiments, the PD-l binding antagonist is an anti-PD-l antibody. In a specific aspect, a PD-l binding antagonist is MDX-1106 (nivolumab). In another specific aspect, a PD-l binding antagonist is MK-3475 (pembrolizumab). In another specific aspect, a PD-l binding antagonist is CT-011 (pidilizumab). In another specific aspect, a PD-l binding antagonist is AMP-224.

[0072] The term“PD-L1 binding antagonist” refers to a molecule that decreases, blocks, inhibits, abrogates or interferes with signal transduction resulting from the interaction of PD-L1 with either one or more of its binding partners, such as PD-l or B7-1. In some embodiments, a PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partners. In a specific aspect, the PD-L1 binding antagonist inhibits binding of PD- Ll to PD-l and/or B7-1. In some embodiments, the PD-L1 binding antagonists include anti- PD-L1 antibodies, antigen binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that decrease, block, inhibit, abrogate or interfere with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners, such as PD-l or B7-1. In one embodiment, a PD-L1 binding antagonist reduces the negative co- stimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes mediated signaling through PD-L1 so as to render a dysfunctional T-cell less dysfunctional (e.g., enhancing effector responses to antigen recognition). In some embodiments, a PD-L1 binding antagonist is an anti-PD-Ll antibody. In a specific aspect, an anti-PD-Ll antibody is YW243.55.S70. In another specific aspect, an anti-PD-Ll antibody is MDX-1105. In still another specific aspect, an anti-PD-Ll antibody is MPDL3280A. In still another specific aspect, an anti-PD-Ll antibody is MEDI4736.

[0073] The term“PD-L2 binding antagonist” refers to a molecule that decreases, blocks, inhibits, abrogates or interferes with signal transduction resulting from the interaction of PD-L2 with either one or more of its binding partners, such as PD-L In some embodiments, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to one or more of its binding partners. In a specific aspect, the PD-L2 binding antagonist inhibits binding of PD- L2 to PD-l. In some embodiments, the PD-L2 antagonists include anti-PD-L2 antibodies, antigen binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that decrease, block, inhibit, abrogate or interfere with signal transduction resulting from the interaction of PD-L2 with either one or more of its binding partners, such as PD-l. In one embodiment, a PD-L2 binding antagonist reduces the negative co-stimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes mediated signaling through PD-L2 so as render a dysfunctional T-cell less dysfunctional (e.g., enhancing effector responses to antigen recognition). In some embodiments, a PD-L2 binding antagonist is an immunoadhesin.

[0074] An“immune checkpoint inhibitor” refers to any compound inhibiting the function of an immune checkpoint protein. Inhibition includes reduction of function and full blockade. In particular the immune checkpoint protein is a human immune checkpoint protein. Thus, the immune checkpoint protein inhibitor in particular is an inhibitor of a human immune checkpoint protein.

[0075] An“extracellular matrix degradative protein” or“extracellular matrix degrading protein” refers any protein which acts on the integrity of the cell matrix, in particular exerting a total or partial degrading or destabilizing action on at least one of the constituents of the said matrix or on the bonds which unite these various constituents.

[0076] An“abscopal effect” is referred to herein as a shrinking of tumors outside the scope of the localized treatment of a tumor. For example, localized treatment with a vims composition provided herein in combination with systemic treatment with an immune checkpoint therapy can result in an abscopal effect at distant tumors that is not injected with the vims composition.

II. Virus Composition

[0077] Certain embodiments of the present disclosure concern a replication-deficient human type 5 adenovirus (Ad5) encoding for expression of the p53 tumor suppressor genes utilized to prepare the p53 dendritic cell vaccine. The construction, properties and purification of the vector have been reported for Ad5/CMV p53 (Zhang 1994; incorporated herein by reference in its entirety). In some aspects, the p53 dendritic cell vaccine is prepared using an adenoviral p53 vector and administered with at least one immune checkpoint inhibitor (U.S. Patent No. 8,668,905; International Patent Publication No. PCTUS2016025912; both of which are incorporated herein by reference in their entirety).

III. Nucleic Acids

[0078] In some aspects, the subject is further administered a tumor suppressor immune gene therapy (PCT/US2016/060833; incorporated herein by reference in its entirety). In some aspects, the subject is further administered additional viral and/or non-viral gene therapies (PCT/US2017/065861 incorporated herein by reference in its entirety).

[0079] In some embodiments, a subject is administered a tumor suppressor therapy, such as a p53 and/or MDA-7 therapy. The nucleic acids encoding p53 and/or MDA-7 may be provided in various methods known in the art.

[0080] In some aspects, the p53 and MDA-7 tumor suppressor therapies incorporate nucleic acid variants to increase their activities. In certain aspects, the variant tumor suppressor nucleic acids are negative regulation-resistant p53 variants (Yun et al., 2012; incorporated herein by reference in its entirety).

[0081] A nucleic acid may be made by any technique known to one of ordinary skill in the art. Non-limiting examples of a synthetic nucleic acid, particularly a synthetic oligonucleotide, include a nucleic acid made by in vitro chemical synthesis using phosphotriester, phosphite or phosphoramidite chemistry and solid phase techniques such as described in EP 266,032, or via deoxynucleoside H-phosphonate intermediates as described by Froehler et al., 1986, and U.S. Patent No. 5,705,629. A non-limiting example of enzymatically produced nucleic acid includes one produced by enzymes in amplification reactions such as PCR™ (see for example, U.S. Patent No. 4,683,202 and U.S. Patent No. 4,682,195), or the synthesis of oligonucleotides described in U.S. Patent No. 5,645,897. A non-limiting example of a biologically produced nucleic acid includes recombinant nucleic acid production in living cells, such as recombinant DNA vector production in bacteria (see for example, Sambrook et al. 1989).

[0082] The nucleic acid(s), regardless of the length of the sequence itself, may be combined with other nucleic acid sequences, including but not limited to, promoters, enhancers, polyadenylation signals, restriction enzyme sites, multiple cloning sites, coding segments, and the like, to create one or more nucleic acid construct(s). The overall length may vary considerably between nucleic acid constructs. Thus, a nucleic acid segment of almost any length may be employed, with the total length preferably being limited by the ease of preparation or use in the intended recombinant nucleic acid protocol.

A. Nucleic Acid Delivery by Expression Vector

[0083] Vectors provided herein are designed, primarily, to express a therapeutic gene (e.g., an immune stimulatory gene such as IL-12 and/or a prodrug converting gene like cytochrome p450 and/or a viral derived lysis promoting gene like ADP) and/or extracellular matrix degradative gene (e.g., relaxin) under the control of regulated eukaryotic promoters (i.e., constitutive, inducible, repressable, tissue- specific). In some aspects, the therapeutic genes may be co-expressed in a vector. In another aspect, the therapeutic genes may be co-expressed with an extracellular matrix degradative gene. Also, the vectors may contain a selectable marker if, for no other reason, to facilitate their manipulation in vitro.

[0084] One of skill in the art would be well-equipped to construct a vector through standard recombinant techniques (see, for example, Sambrook et al, 2001 and Ausubel et al, 1996, both incorporated herein by reference). Vectors include but are not limited to, plasmids, cosmids, viruses (bacteriophage, animal viruses, and plant viruses), and artificial chromosomes (e.g., YACs), such as retroviral vectors (e.g. derived from Moloney murine leukemia vims vectors (MoMLV), MSCV, SFFV, MPSV, SNV etc), lentiviral vectors (e.g. derived from HIV- 1, HIV-2, SIV, BIV, FIV etc.), adenoviral (Ad) vectors including replication competent, replication deficient and gutless forms thereof, adeno-associated viral (AAV) vectors, simian vims 40 (SV-40) vectors, bovine papilloma virus vectors, Epstein-Barr vims vectors, herpes vims vectors, vaccinia virus vectors, Harvey murine sarcoma virus vectors, murine mammary tumor vims vectors, Rous sarcoma vims vectors, parvovirus vectors, polio vims vectors, vesicular stomatitis vims vectors, maraba vims vectors and group B adenovims enadenotucirev vectors.

1. Viral Vectors

[0085] Viral vectors encoding a therapeutic gene may be provided in certain aspects of the present disclosure. In generating recombinant viral vectors, non-essential genes are typically replaced with a gene or coding sequence for a heterologous (or non-native) protein. A viral vector is a kind of expression construct that utilizes viral sequences to introduce nucleic acid and possibly proteins into a cell. The ability of certain viruses to infect cells or enter cells via receptor mediated- endocytosis, and to integrate into host cell genomes and express viral genes stably and efficiently have made them attractive candidates for the transfer of foreign nucleic acids into cells (e.g., mammalian cells). Non- limiting examples of vims vectors that may be used to deliver a nucleic acid of certain aspects of the present invention are described below.

[0086] Lentiviruses are complex retroviruses, which, in addition to the common retroviral genes gag, pol, and env, contain other genes with regulatory or structural function. Lentiviral vectors are well known in the art (see, for example, Naldini et al, 1996; Zufferey et al, 1997; Blomer et al, 1997; U.S. Patents 6,013,516 and 5,994,136).

[0087] Recombinant lentiviral vectors are capable of infecting non-dividing cells and can be used for both in vivo and ex vivo gene transfer and expression of nucleic acid sequences. For example, recombinant lentivirus capable of infecting a non-dividing cell— wherein a suitable host cell is transfected with two or more vectors carrying the packaging functions, namely gag, pol and env, as well as rev and tat— is described in U.S. Patent 5,994,136, incorporated herein by reference. a. Adenoviral Vector

[0088] One method for delivery of the tumor suppressor and/or extracellular matrix degradative gene involves the use of an adenovirus expression vector. Although adenovirus vectors are known to have a low capacity for integration into genomic DNA, this feature is counterbalanced by the high efficiency of gene transfer afforded by these vectors. Adenovirus expression vectors include constructs containing adenovirus sequences sufficient to (a) support packaging of the construct and (b) to ultimately express a recombinant gene construct that has been cloned therein.

[0089] Adenovirus growth and manipulation is known to those of skill in the art and exhibits broad host range in vitro and in vivo. This group of viruses can be obtained in high titers, e.g., 109-1011 plaque-forming units per ml, and they are highly infective. The life cycle of adenovirus does not require integration into the host cell genome. The foreign genes delivered by adenovirus vectors are episomal and, therefore, have low genotoxicity to host cells. No side effects have been reported in studies of vaccination with wild-type adenovirus (Couch et al., 1963; Top et al., 1971), demonstrating their safety and therapeutic potential as in vivo gene transfer vectors. [0090] Knowledge of the genetic organization of adenovirus, a 36 kb, linear, double- stranded DNA virus, allows substitution of large pieces of adenoviral DNA with foreign sequences up to 7 kb (Grunhaus and Horwitz, 1992). In contrast to retrovirus, the adenoviral infection of host cells does not result in chromosomal integration because adenoviral DNA can replicate in an episomal manner without potential genotoxicity. Also, adenoviruses are structurally stable, and no genome rearrangement has been detected after extensive amplification.

[0091] Adenovirus is particularly suitable for use as a gene transfer vector because of its mid-sized genome, ease of manipulation, high titer, wide target-cell range and high infectivity. Both ends of the viral genome contain 100-200 base pair inverted repeats (ITRs), which are cis elements necessary for viral DNA replication and packaging. The early (E) and late (L) regions of the genome contain different transcription units that are divided by the onset of viral DNA replication. The El region (E1A and E1B) encodes proteins responsible for the regulation of transcription of the viral genome and a few cellular genes. The expression of the E2 region (E2A and E2B) results in the synthesis of the proteins for viral DNA replication. These proteins are involved in DNA replication, late gene expression and host cell shut-off (Renan, 1990). The products of the late genes, including the majority of the viral capsid proteins, are expressed only after significant processing of a single primary transcript issued by the major late promoter (MLP). The MLP, (located at 16.8 m.u.) is particularly efficient during the late phase of infection, and all the mRNA's issued from this promoter possess a 5'- tripartite leader (TPL) sequence which makes them particular mRNA's for translation.

[0092] A recombinant adenovirus provided herein can be generated from homologous recombination between a shuttle vector and provirus vector. Due to the possible recombination between two proviral vectors, wild-type adenovirus may be generated from this process. Therefore, a single clone of virus is isolated from an individual plaque and its genomic structure is examined.

[0093] The adenovirus vector may be replication competent, replication defective, or conditionally defective, the nature of the adenovirus vector is not believed to be crucial to the successful practice of the invention. The adenovirus may be of any of the 42 different known serotypes or subgroups A-F. Adenovirus type 5 of subgroup C is the particular starting material in order to obtain the conditional replication-defective adenovirus vector for use in the present invention. This is because Adenovirus type 5 is a human adenovirus about which a great deal of biochemical and genetic information is known, and it has historically been used for most constructions employing adenovirus as a vector. However, other serotypes of adenovirus may be similarly utilized.

[0094] Nucleic acids can be introduced to adenoviral vectors as a position from which a coding sequence has been removed. For example, a replication defective adenoviral vector can have the El -coding sequences removed. The polynucleotide encoding the gene of interest may also be inserted in lieu of the deleted E3 region in E3 replacement vectors as described by Karlsson et al. (1986) or in the E4 region where a helper cell line or helper vims complements the E4 defect.

[0095] Generation and propagation of replication deficient adenovirus vectors can be performed with helper cell lines. One unique helper cell line, designated 293, was transformed from human embryonic kidney cells by Ad5 DNA fragments and constitutively expresses El proteins (Graham et al. , 1977). Since the E3 region is dispensable from the adenovirus genome (Jones and Shenk, 1978), adenovirus vectors, with the help of 293 cells, carry foreign DNA in either the El, the E3, or both regions (Graham and Prevec, 1991).

[0096] Helper cell lines may be derived from human cells such as human embryonic kidney cells, muscle cells, hematopoietic cells or other human embryonic mesenchymal or epithelial cells. Alternatively, the helper cells may be derived from the cells of other mammalian species that are permissive for human adenovirus. Such cells include, e.g., Vero cells or other monkey embryonic mesenchymal or epithelial cells. As stated above, a particular helper cell line is 293.

[0097] Methods for producing recombinant adenovirus are known in the art, such as U.S. Patent No. 6740320, incorporated herein by reference. Also, Racher et al. (1995) have disclosed improved methods for culturing 293 cells and propagating adenovirus. In one format, natural cell aggregates are grown by inoculating individual cells into 1 liter siliconized spinner flasks (Techne, Cambridge, UK) containing 100-200 ml of medium. Following stirring at 40 rpm, the cell viability is estimated with trypan blue. In another format, Fibra-Cel microcarriers (Bibby Sterlin, Stone, UK) (5 g/l) are employed as follows. A cell inoculum, resuspended in 5 ml of medium, is added to the carrier (50 ml) in a 250 ml Erlenmeyer flask and left stationary, with occasional agitation, for 1 to 4 hours. The medium is then replaced with 50 ml of fresh medium and shaking initiated. For virus production, cells are allowed to grow to about 80% confluence, after which time the medium is replaced (to 25% of the final volume) and adenovirus added at an MOI of 0.05. Cultures are left stationary overnight, following which the volume is increased to 100% and shaking commenced for another 72 hours. b. Retroviral Vector

[0098] Additionally, the viral composition may comprise a retroviral vector. The retroviruses are a group of single-stranded RNA viruses characterized by an ability to convert their RNA to double-stranded DNA in infected cells by a process of reverse-transcription (Coffin, 1990). The resulting DNA then stably integrates into cellular chromosomes as a provirus and directs synthesis of viral proteins. The integration results in the retention of the viral gene sequences in the recipient cell and its descendants. The retroviral genome contains three genes, gag, pol, and env that code for capsid proteins, polymerase enzyme, and envelope components, respectively. A sequence found upstream from the gag gene contains a signal for packaging of the genome into virions. Two long terminal repeat (LTR) sequences are present at the 5' and 3' ends of the viral genome. These contain strong promoter and enhancer sequences and are also required for integration in the host cell genome (Coffin, 1990).

[0099] In order to construct a retroviral vector, a nucleic acid encoding a gene of interest is inserted into the viral genome in the place of certain viral sequences to produce a vims that is replication-defective. In order to produce virions, a packaging cell line containing the gag, pol, and env genes but without the LTR and packaging components is constructed (Mann et al, 1983). When a recombinant plasmid containing a cDNA, together with the retroviral LTR and packaging sequences is introduced into this cell line (by calcium phosphate precipitation for example), the packaging sequence allows the RNA transcript of the recombinant plasmid to be packaged into viral particles, which are then secreted into the culture media (Nicolas and Rubenstein, 1988; Temin, 1986; Mann et al, 1983). The media containing the recombinant retroviruses is then collected, optionally concentrated, and used for gene transfer. Retroviral vectors are able to infect a broad variety of cell types. However, integration and stable expression require the division of host cells (Paskind et al, 1975).

[00100] Concern with the use of defective retrovirus vectors is the potential appearance of wild-type replication-competent vims in the packaging cells. This can result from recombination events in which the intact sequence from the recombinant vims inserts upstream from the gag, pol, env sequence integrated in the host cell genome. However, packaging cell lines are available that should greatly decrease the likelihood of recombination (Markowitz et al, 1988; Hersdorffer el al, 1990). c. Adeno-associated Viral Vector

[00101] Adeno-associated vims (AAV) is an attractive vector system for use in the present disclosure as it has a high frequency of integration and it can infect nondividing cells, thus making it useful for delivery of genes into mammalian cells (Muzyczka, 1992). AAV has a broad host range for infectivity (Tratschin, et al. , 1984; Laughlin, et al. , 1986; Lebkowski, et al , 1988; McLaughlin, et al., 1988), which means it is applicable for use with the present invention. Details concerning the generation and use of rAAV vectors are described in U.S. Patent No. 5,139,941 and U.S. Patent No. 4,797,368.

[00102] AAV is a dependent parvovirus in that it requires coinfection with another vims (either adenovims or a member of the herpes virus family) to undergo a productive infection in cultured cells (Muzyczka, 1992). In the absence of coinfection with helper vims, the wild-type AAV genome integrates through its ends into human chromosome 19 where it resides in a latent state as a provims (Kotin et al., 1990; Samulski et al., 1991). rAAV, however, is not restricted to chromosome 19 for integration unless the AAV Rep protein is also expressed (Shelling and Smith, 1994). When a cell carrying an AAV provirus is superinfected with a helper vims, the AAV genome is "rescued" from the chromosome or from a recombinant plasmid, and a normal productive infection is established (Samulski et al, 1989; McLaughlin et al., 1988; Kotin et al, 1990; Muzyczka, 1992).

[00103] Typically, recombinant AAV (rAAV) vims is made by cotransfecting a plasmid containing the gene of interest flanked by the two AAV terminal repeats (McLaughlin et al, 1988; Samulski et al, 1989; each incorporated herein by reference) and an expression plasmid containing the wild-type AAV coding sequences without the terminal repeats, for example pIM45 (McCarty et al, 1991). The cells are also infected or transfected with adenovims or plasmids carrying the adenovirus genes required for AAV helper function. rAAV vims stocks made in such fashion are contaminated with adenovirus which must be physically separated from the rAAV particles (for example, by cesium chloride density centrifugation). Alternatively, adenovims vectors containing the AAV coding regions or cell lines containing the AAV coding regions and some or all of the adenovims helper genes could be used (Yang et al, 1994; Clark et al, 1995). Cell lines carrying the rAAV DNA as an integrated provims can also be used (Flotte et al, 1995). d. Other Viral Vectors

[00104] Other viral vectors may be employed as constructs in the present disclosure. Vectors derived from viruses such as vaccinia vims (Ridgeway, 1988; Baichwal and Sugden, 1986; Coupar et al , 1988) and herpesviruses may be employed. They offer several attractive features for various mammalian cells (Friedmann, 1989; Ridgeway, 1988; Baichwal and Sugden, 1986; Coupar et al, 1988; Horwich et al, 1990).

[00105] A molecularly cloned strain of Venezuelan equine encephalitis (VEE) vims has been genetically refined as a replication competent vaccine vector for the expression of heterologous viral proteins (Davis et al, 1996). Studies have demonstrated that VEE infection stimulates potent CTL responses and has been suggested that VEE may be an extremely useful vector for immunizations (Caley et al, 1997).

[00106] In further embodiments, the nucleic acid is housed within an infective vims that has been engineered to express a specific binding ligand. The vims particle will thus bind specifically to the cognate receptors of the target cell and deliver the contents to the cell. A novel approach designed to allow specific targeting of retrovims vectors was recently developed based on the chemical modification of a retrovirus by the chemical addition of lactose residues to the viral envelope. This modification can permit the specific infection of hepatocytes via sialoglycoprotein receptors.

[00107] For example, targeting of recombinant retrovimses was designed in which biotinylated antibodies against a retroviral envelope protein and against a specific cell receptor were used. The antibodies were coupled via the biotin components by using streptavidin (Roux et al, 1989). Using antibodies against major histocompatibility complex class I and class II antigens, they demonstrated the infection of a variety of human cells that bore those surface antigens with an ecotropic vims in vitro (Roux et al, 1989).

2. Regulatory Elements

[00108] Expression cassettes included in vectors useful in the present disclosure in particular contain (in a 5'-to-3' direction) a eukaryotic transcriptional promoter operably linked to a protein-coding sequence, splice signals including intervening sequences, and a transcriptional termination/polyadenylation sequence. The promoters and enhancers that control the transcription of protein encoding genes in eukaryotic cells are composed of multiple genetic elements. The cellular machinery is able to gather and integrate the regulatory information conveyed by each element, allowing different genes to evolve distinct, often complex patterns of transcriptional regulation. A promoter used in the context of the present invention includes constitutive, inducible, and tissue-specific promoters. a. Promoter/Enhancers

[00109] The expression constructs provided herein comprise a promoter to drive expression of the tumor suppressor and/or extracellular matrix degradative gene. A promoter generally comprises a sequence that functions to position the start site for RNA synthesis. The best known example of this is the TATA box, but in some promoters lacking a TATA box, such as, for example, the promoter for the mammalian terminal deoxynucleotidyl transferase gene and the promoter for the SV40 late genes, a discrete element overlying the start site itself helps to fix the place of initiation. Additional promoter elements regulate the frequency of transcriptional initiation. Typically, these are located in the region 30110 bp- upstream of the start site, although a number of promoters have been shown to contain functional elements downstream of the start site as well. To bring a coding sequence“under the control of’ a promoter, one positions the 5' end of the transcription initiation site of the transcriptional reading frame“downstream” of (/._<?., 3' of) the chosen promoter. The“upstream” promoter stimulates transcription of the DNA and promotes expression of the encoded RNA.

[00110] The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or moved relative to one another. In the tk promoter, the spacing between promoter elements can be increased to 50 bp apart before activity begins to decline. Depending on the promoter, it appears that individual elements can function either cooperatively or independently to activate transcription. A promoter may or may not be used in conjunction with an“enhancer,” which refers to a ex acting regulatory sequence involved in the transcriptional activation of a nucleic acid sequence.

[00111] A promoter may be one naturally associated with a nucleic acid sequence, as may be obtained by isolating the 5' non-coding sequences located upstream of the coding segment and/or exon. Such a promoter can be referred to as“endogenous.” Similarly, an enhancer may be one naturally associated with a nucleic acid sequence, located either downstream or upstream of that sequence. Alternatively, certain advantages will be gained by positioning the coding nucleic acid segment under the control of a recombinant or heterologous promoter, which refers to a promoter that is not normally associated with a nucleic acid sequence in its natural environment. A recombinant or heterologous enhancer refers also to an enhancer not normally associated with a nucleic acid sequence in its natural environment. Such promoters or enhancers may include promoters or enhancers of other genes, and promoters or enhancers isolated from any other vims, or prokaryotic or eukaryotic cell, and promoters or enhancers not “naturally occurring,” i.e., containing different elements of different transcriptional regulatory regions, and/or mutations that alter expression. For example, promoters that are most commonly used in recombinant DNA construction include the b lactamase (penicillinase), lactose and tryptophan (trp-) promoter systems. In addition to producing nucleic acid sequences of promoters and enhancers synthetically, sequences may be produced using recombinant cloning and/or nucleic acid amplification technology, including PCR™, in connection with the compositions disclosed herein (see U.S. Patent Nos. 4,683,202 and 5,928,906, each incorporated herein by reference). Furthermore, it is contemplated that the control sequences that direct transcription and/or expression of sequences within non nuclear organelles such as mitochondria, chloroplasts, and the like, can be employed as well.

[00112] Naturally, it will be important to employ a promoter and/or enhancer that effectively directs the expression of the DNA segment in the organelle, cell type, tissue, organ, or organism chosen for expression. Those of skill in the art of molecular biology generally know the use of promoters, enhancers, and cell type combinations for protein expression, (see, for example Sambrook et al. 1989, incorporated herein by reference). The promoters employed may be constitutive, tissue-specific, inducible, and/or useful under the appropriate conditions to direct high level expression of the introduced DNA segment, such as is advantageous in the large-scale production of recombinant proteins and/or peptides. The promoter may be heterologous or endogenous.

[00113] Additionally, any promoter/enhancer combination (as per, for example, the Eukaryotic Promoter Data Base EPDB, through world wide web at epd.isb-sib.ch/) could also be used to drive expression. Use of a T3, T7 or SP6 cytoplasmic expression system is another possible embodiment. Eukaryotic cells can support cytoplasmic transcription from certain bacterial promoters if the appropriate bacterial polymerase is provided, either as part of the delivery complex or as an additional genetic expression construct.

[00114] Non-limiting examples of promoters include early or late viral promoters, such as, SV40 early or late promoters, cytomegalovirus (CMV) immediate early promoters, Rous Sarcoma Vims (RSV) early promoters; eukaryotic cell promoters, such as, e. g. , beta actin promoter (Ng, 1989; Quitsche et al , 1989), GADPH promoter (Alexander et al , 1988, Ercolani et al , 1988), metallothionein promoter (Karin et al, 1989; Richards et al, 1984); and concatenated response element promoters, such as cyclic AMP response element promoters (ere), serum response element promoter (sre), phorbol ester promoter (TPA) and response element promoters (tre) near a minimal TATA box. It is also possible to use human growth hormone promoter sequences (e.g., the human growth hormone minimal promoter described at Genbank, accession no. X05244, nucleotide 283-341) or a mouse mammary tumor promoter (available from the ATCC, Cat. No. ATCC 45007). In certain embodiments, the promoter is CMV IE, dectin-l, dectin-2, human CDllc, F4/80, SM22, RSV, SV40, Ad MLP, beta-actin, MHC class I or MHC class II promoter, however any other promoter that is useful to drive expression of the therapeutic gene is applicable to the practice of the present invention.

[00115] In certain aspects, methods of the disclosure also concern enhancer sequences, i.e., nucleic acid sequences that increase a promoter’s activity and that have the potential to act in cis, and regardless of their orientation, even over relatively long distances (up to several kilobases away from the target promoter). However, enhancer function is not necessarily restricted to such long distances as they may also function in close proximity to a given promoter. b. Initiation Signals and Linked Expression

[00116] A specific initiation signal also may be used in the expression constructs provided in the present disclosure for efficient translation of coding sequences. These signals include the ATG initiation codon or adjacent sequences. Exogenous translational control signals, including the ATG initiation codon, may need to be provided. One of ordinary skill in the art would readily be capable of determining this and providing the necessary signals. It is well known that the initiation codon must be“in-frame” with the reading frame of the desired coding sequence to ensure translation of the entire insert. The exogenous translational control signals and initiation codons can be either natural or synthetic. The efficiency of expression may be enhanced by the inclusion of appropriate transcription enhancer elements.

[00117] In certain embodiments, the use of internal ribosome entry sites (IRES) elements are used to create multigene, or polycistronic, messages. IRES elements are able to bypass the ribosome scanning model of 5' methylated Cap dependent translation and begin translation at internal sites (Pelletier and Sonenberg, 1988). IRES elements from two members of the picornavirus family (polio and encephalomyocarditis) have been described (Pelletier and Sonenberg, 1988), as well an IRES from a mammalian message (Macejak and Sarnow, 1991). IRES elements can be linked to heterologous open reading frames. Multiple open reading frames can be transcribed together, each separated by an IRES, creating polycistronic messages. By virtue of the IRES element, each open reading frame is accessible to ribosomes for efficient translation. Multiple genes can be efficiently expressed using a single promoter/enhancer to transcribe a single message (see U.S. Patent Nos. 5,925,565 and 5,935,819, each herein incorporated by reference).

[00118] Additionally, certain 2A sequence elements could be used to create linked- or co-expression of genes in the constructs provided in the present disclosure. For example, cleavage sequences could be used to co-express genes by linking open reading frames to form a single cistron. An exemplary cleavage sequence is the F2A (Foot-and-mouth diease vims 2A) or a“2A-like” sequence (e.g., Thosea asigna vims 2A; T2A) (Minskaia and Ryan, 2013). c. Origins of Replication

In order to propagate a vector in a host cell, it may contain one or more origins of replication sites (often termed“ori”), for example, a nucleic acid sequence corresponding to oriP of EBV as described above or a genetically engineered oriP with a similar or elevated function in programming, which is a specific nucleic acid sequence at which replication is initiated. Alternatively, a replication origin of other extra-chromosomally replicating virus as described above or an autonomously replicating sequence (ARS) can be employed.

3. Selection and Screenable Markers

[00119] In some embodiments, cells containing a construct of the present disclosure may be identified in vitro or in vivo by including a marker in the expression vector. Such markers would confer an identifiable change to the cell permitting easy identification of cells containing the expression vector. Generally, a selection marker is one that confers a property that allows for selection. A positive selection marker is one in which the presence of the marker allows for its selection, while a negative selection marker is one in which its presence prevents its selection. An example of a positive selection marker is a drug resistance marker.

[00120] Usually the inclusion of a drug selection marker aids in the cloning and identification of transformants, for example, genes that confer resistance to neomycin, puromycin, hygromycin, DHFR, GPT, zeocin and histidinol are useful selection markers. In addition to markers conferring a phenotype that allows for the discrimination of transformants based on the implementation of conditions, other types of markers including screenable markers such as GFP, whose basis is colorimetric analysis, are also contemplated. Alternatively, screenable enzymes as negative selection markers such as herpes simplex vims thymidine kinase (tk) or chloramphenicol acetyltransferase (CAT) may be utilized. One of skill in the art would also know how to employ immunologic markers, possibly in conjunction with FACS analysis. The marker used is not believed to be important, so long as it is capable of being expressed simultaneously with the nucleic acid encoding a gene product. Further examples of selection and screenable markers are well known to one of skill in the art.

B. Other Methods of Nucleic Acid Delivery

[00121] In addition to viral delivery of the nucleic acids encoding the therapeutic gene and/or extracellular matrix degradative gene, the following are additional methods of recombinant gene delivery to a given host cell and are thus considered in the present disclosure. Thus, other forms of gene therapy may be combined with the therapeutic viral compositions including gene editing methods such as meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector-based nucleases (TALEN), and the CRISPR-Cas system.

[00122] Introduction of a nucleic acid, such as DNA or RNA, may use any suitable methods for nucleic acid delivery for transformation of a cell, as described herein or as would be known to one of ordinary skill in the art. Such methods include, but are not limited to, direct delivery of DNA such as by ex vivo transfection (Wilson et al, 1989, Nabel et al, 1989), by injection (U.S. Patent Nos. 5,994,624, 5,981,274, 5,945,100, 5,780,448, 5,736,524, 5,702,932, 5,656,610, 5,589,466 and 5,580,859, each incorporated herein by reference), including microinjection (Harland and Weintraub, 1985; U.S. Patent No. 5,789,215, incorporated herein by reference); by electroporation (U.S. Patent No. 5,384,253, incorporated herein by reference; Tur-Kaspa et al, 1986; Potter et al, 1984); by calcium phosphate precipitation (Graham and Van Der Eb, 1973; Chen and Okayama, 1987; Rippe et al., 1990); by using DEAE-dextran followed by polyethylene glycol (Gopal, 1985); by direct sonic loading (Fechheimer et al., 1987); by liposome mediated transfection (Nicolau and Sene, 1982; Fraley et al, 1979; Nicolau el al, 1987; Wong el al, 1980; Kaneda et al, 1989; Kato et al, 1991) and receptor-mediated transfection (Wu and Wu, 1987; Wu and Wu, 1988); by microprojectile bombardment (PCT Application Nos. WO 94/09699 and 95/06128; U.S. Patent Nos. 5,610,042; 5,322,783 5,563,055, 5,550,318, 5,538,877 and 5,538,880, and each incorporated herein by reference); by agitation with silicon carbide fibers (Kaeppler et al, 1990; U.S. Patent Nos. 5,302,523 and 5,464,765, each incorporated herein by reference); by Agrobacteriummediated transformation (U.S. Patent Nos. 5,591,616 and 5,563,055, each incorporated herein by reference); by desiccation/- inhibitionmediated DNA uptake (-Potrykus et al, 1985), and any combination of such methods. Through the application of techniques such as these, organelle(s), cell(s), tissue(s) or organism(s) may be stably or transiently transformed.

1. Electroporation

[00123] In certain particular embodiments of the present disclosure, the gene construct is introduced into target hyperproliferative cells via electroporation. Electroporation involves the exposure of cells (or tissues) and DNA (or a DNA complex) to a high-voltage electric discharge.

[00124] Transfection of eukaryotic cells using electroporation has been quite successful. Mouse pre-B lymphocytes have been transfected with human kappa- immunoglobulin genes (Potter et al, 1984), and rat hepatocytes have been transfected with the chloramphenicol acetyltransferase gene (Tur-Kaspa et al, 1986) in this manner.

[00125] It is contemplated that electroporation conditions for hyperproliferative cells from different sources may be optimized. One may particularly wish to optimize such parameters as the voltage, the capacitance, the time and the electroporation media composition. The execution of other routine adjustments will be known to those of skill in the art. See e.g. , Hoffman, 1999; Heller et al , 1996.

2. Lipid-Mediated Transformation

[00126] In a further embodiment, the tumor suppressor and/or extracellular matrix degradative gene may be entrapped in a liposome or lipid formulation. Liposomes are vesicular structures characterized by a phospholipid bilayer membrane and an inner aqueous medium. Multilamellar liposomes have multiple lipid layers separated by aqueous medium. They form spontaneously when phospholipids are suspended in an excess of aqueous solution. The lipid components undergo self-rearrangement before the formation of closed structures and entrap water and dissolved solutes between the lipid bilayers (Ghosh and Bachhawat, 1991). Also contemplated is a gene construct complexed with Lipofectamine (Gibco BRL). [00127] Lipid-mediated nucleic acid delivery and expression of foreign DNA in vitro has been very successful (Nicolau and Sene, 1982; Fraley et al, 1979; Nicolau et al, 1987). Wong et al. (1980) demonstrated the feasibility of lipid-mediated delivery and expression of foreign DNA in cultured chick embryo, HeLa and hepatoma cells.

[00128] Lipid based non- viral formulations provide an alternative to adenoviral gene therapies. Although many cell culture studies have documented lipid based non- viral gene transfer, systemic gene delivery via lipid-based formulations has been limited. A major limitation of non-viral lipid-based gene delivery is the toxicity of the cationic lipids that comprise the non-viral delivery vehicle. The in vivo toxicity of liposomes partially explains the. discrepancy between in vitro and in vivo gene transfer results. Another factor contributing to this contradictory data is the difference in lipid vehicle stability in the presence and absence of serum proteins. The interaction between lipid vehicles and serum proteins has a dramatic impact on the stability characteristics of lipid vehicles (Yang and Huang, 1997). Cationic lipids attract and bind negatively charged serum proteins. Lipid vehicles associated with serum proteins are either dissolved or taken up by macrophages leading to their removal from circulation. Current in vivo lipid delivery methods use subcutaneous, intradermal, intratumoral, or intracranial injection to avoid the toxicity and stability problems associated with cationic lipids in the circulation. The interaction of lipid vehicles and plasma proteins is responsible for the disparity between the efficiency of in vitro (Felgner et al, 1987) and in vivo gene transfer (Zhu el al., 1993; Philip et al , 1993; Solodin et al, 1995; Liu et al, 1995; Thierry et al , 1995; Tsukamoto et al , 1995; Aksentijevich et al , 1996).

[00129] Advances in lipid formulations have improved the efficiency of gene transfer in vivo (Templeton et al 1997; WO 98/07408). A novel lipid formulation composed of an equimolar ratio of l,2-bis(oleoyloxy)-3-(trimethyl ammonio)propane (DOTAP) and cholesterol significantly enhances systemic in vivo gene transfer, approximately 150 fold. The DOTAP:cholesterol lipid formulation forms unique structure termed a "sandwich liposome". This formulation is reported to "sandwich" DNA between an invaginated bi-layer or 'vase' structure. Beneficial characteristics of these lipid structures include a positive p, colloidal stabilization by cholesterol, two-dimensional DNA packing and increased serum stability. Patent Application Nos. 60/135,818 and 60/133,116 discuss formulations that may be used with the present invention. [00130] The production of lipid formulations often is accomplished by sonication or serial extrusion of liposomal mixtures after (I) reverse phase evaporation (II) dehydration-rehydration (III) detergent dialysis and (IV) thin film hydration. Once manufactured, lipid structures can be used to encapsulate compounds that are toxic (chemotherapeutics) or labile (nucleic acids) when in circulation. Lipid encapsulation has resulted in a lower toxicity and a longer serum half-life for such compounds (Gabizon et al , 1990). Numerous disease treatments are using lipid-based gene transfer strategies to enhance conventional or establish novel therapies, in particular therapies for treating hyperproliferative diseases.

IV. Preferential CD123/CD132 Agonists

[00131] In certain aspects, the subject is administered at least one CD122/CD132 agonist, such as a CD122/CD132 agonist that preferentially binds to the CD122/CD132 receptor complex and has lower affinity binding for CD25 or the ILl5a receptor. The CD 122/CD 132 may be selected from a genetically engineered IL-22 mutein that has a modified amino acid sequence compared to wild type IL2 (US 2017/0044229; incorporated by reference in its entirety). In certain aspects, the preferential CD122/CD132 agonist is an IL-2/anti-IL-2 monoclonal antibody immune complex (US20170183403A1 ; incorporated by reference in its entirety), or a genetically engineered IL-2 mutein that has a modified amino acid sequence compared to wild type IL-2 combined with an anti-IL2 monoclonal antibody immune complex (W02014100014A1; incorporated by reference in its entirety), a PEGylated form of IL2 like NKTR-214 (Charych et al, 2016), an ILl5/anti-ILl5 monoclonal antibody immune complex, an IL15/IL15 Receptor a-IgGl-Fc (ILl5/ILl5Ra-IgGl-Fc) immune complex (US20060257361A1, EP2724728A1 and Dubois et al, 2008), a genetically engineered IL-15 mutein that has a modified amino acid sequence compared to wild type IL-15 combined with an ILl5Ra-IgGl-Fc immune complex (US20070160578; incorporated herein in its entirety), or a PEGylated form of IL15 with preferential binding to CD122/CD132. In some embodiments, more than one CD122/CD132 agonist are utilized.

[00132] Additional representative CD122/CD132 agonists for use in the present disclosure include but are not limited to the agents listed in Table 1 below.

Abbreviations: MAb=monoclonal antibody; Ab=antibody; r=recombinant; h=human; Ra=Receptor Alpha.

V. Oncolytic Viruses

[00133] In some aspects, the present disclosure comprises administration of at least one oncolytic virus. In some aspects, the oncolytic vims is engineered to express p53, MDA-7, IL-12, TGF-b inhibitor, IL-10 inhibitor and/or heat shock protein. In certain aspects, the oncolytic vims is a single- or double-stranded DNA vims, RNA vims, adenovirus, adeno- associated vims, retrovims, lentivims, herpes virus, pox virus, vaccinia vims, vesicular stomatitis virus, polio virus, Newcastle’s Disease vims, Epstein-Barr virus, influenza vims, reoviruses, myxoma vims, maraba virus, rhabdovims, enadenotucirev or coxsackie virus. In some aspects, the oncolytic vims is engineered to express a cytokine, such as granulocyte- macrophage colony- stimulating factor (GM-CSF) or IL-12. In some aspects, the oncolytic virus is further defined as talimogene laherparepvec (T-VEC). In some aspects, the oncolytic adenoviral vector is derived from a modified TERT Promoter Oncolytic Adenovims (US Patent No. 8,067,567; incorporated herein by reference in its entirety) and/or the HRE-E2F-TERT Hybrid Promoter Oncolytic Adeno virus (PCT/KR2011/004693; incorporated herein by reference in its entirety) and/or an adenovims with a modified Ela regulatory sequence wherein at least one Pea3 binding site, or a functional portion thereof, is deleted with an Elb-l9K clone insertion site (EP2403951A2; incorporated herein by reference in its entirety) which may all be modified to express therapeutic genes. In some aspects, the oncolytic adenoviral vector is derived from Elb deleted oncolytic adenovimses (Yu and Fang, 2007; Li, 2009; both incorporated by reference in their entirety).

[00134] Exemplary oncolytic viruses include, but are not limited to, Ad5- y CD/mutTKS R39rep -hIL 12 , Cavatak™, CG0070, DNX-2401, G207, HF10, IMLYGIC™, JX-594, MG1-MA3, MV-NIS, OBP-301, Reolysin®, Toca 511, Oncorine, RIGVIR, an adenovims overexpressing the adenoviral death protein (ADP) as described in US 7589069 Bl incorporated by reference in its entirety, such as VirRx007, an N1L deleted vaccinia virus expressing IL12 as described in PCT/GB2015/051023 incorporated by reference in its entirety. Other exemplary oncolytic viruses are described, for example, in International Patent Publication Nos. WO2015/027163, WO2014/138314, W02014/047350, and

WO2016/009017; all incorporated herein by reference.

[00135] In a particular aspects, the oncolytic viral agent is talimogene laherparepvec (T-VEC) which is an oncolytic herpes simplex virus genetically engineered to express GM-CSF. Talimogene laherparepvec, HSV-l [strain JS1] ICP34.5-/ICP47-/hGM- CSF, (previously known as OncoVEX^{GM CSF}), is an intratumorally delivered oncolytic immunotherapy comprising an immune-enhanced HSV-l that selectively replicates in solid tumors. (Lui et al., 2003; U.S. Patent No. 7,223,593 and U.S. Patent No. 7,537,924; incorporated herein by reference). In October 2015, the US FDA approved T-VEC, under the brand name IMLYGIC™, for the treatment of melanoma in patients with inoperable tumors. The characteristics and methods of administration of T-VEC are described in, for example, the IMLYGIC™ package insert (Amgen, 2015) and U.S. Patent Publication No. US2015/0202290; both incorporated herein by reference. For example, talimogene laherparepvec is typically administered by intratumoral injection into injectable cutaneous, subcutaneous, and nodal tumors at a dose of up to 4.0 ml of 10⁶ plaque forming unit/mL (PFU/mL) at day 1 of week 1 followed by a dose of up to 4.0 ml of 10⁸ PFU/mL at day 1 of week 4, and every 2 weeks (± 3 days) thereafter. The recommended volume of talimogene laherparepvec to be injected into the tumor(s) is dependent on the size of the tumor(s) and should be determined according to the injection volume guideline. While T-VEC has demonstrated clinical activity in melanoma patients, many cancer patients either do not respond or cease responding to T-VEC treatment. In one embodiment, the p53 and/or MDA-7 nucleic acids and the at least one CD 122/CD 132 agonist may be administered after, during or before T-VEC therapy, such as to reverse treatment resistance.

[00136] In some embodiments, Elb deleted oncolytic adenoviruses are combined with at least one preferential CD122/CD132 agonist and at least one immune checkpoint inhibitor. Exemplary Elb deleted oncolytic adenoviruses are H101 (Oncorine), Onyx 015 or H103 which expresses the heat shock protein 70 (HSP70) or the oncolytic adenovirus H102 in which expression of the Ad Ela gene is driven by the alpha-fetoprotein (AFP) promoter resulting in preferential replication in hepatocellular carcinoma and other AFP overexpressing cancers compared to normal cells (Yu and Fang, 2007; Li, 2009; both incorporated by reference in their entirety). VI. Immune Checkpoint Inhibitors

[00137] In certain embodiments, the present disclosure provides methods of combining p53 dendritic cell vaccine and CD122/CD132 treatment with immune checkpoint inhibitors. Immune checkpoints are molecules in the immune system that either turn up a signal (e.g., co-stimulatory molecules) or turn down a signal. Inhibitory checkpoint molecules that may be targeted by immune checkpoint blockade include adenosine A2A receptor (A2AR), B7-H3 (also known as CD276), B and T lymphocyte attenuator (BTLA), cytotoxic T- lymphocyte-associated protein 4 (CTLA-4, also known as CD152), indoleamine 2,3- dioxygenase (IDO), killer-cell immunoglobulin (KIR), lymphocyte activation gene-3 (LAG3), programmed death 1 (PD-l), T-cell immunoglobulin domain and mucin domain 3 (TIM-3) and V-domain Ig suppressor of T cell activation (VISTA). In particular, the immune checkpoint inhibitors target the PD-l axis and/or CTLA-4.

[00138] The immune checkpoint inhibitors may be drugs such as small molecules, recombinant forms of ligand or receptors, or, in particular, are antibodies, such as human antibodies (e.g., International Patent Publication W02015016718; Pardoll, 2012; both incorporated herein by reference). Known inhibitors of the immune checkpoint proteins or analogs thereof may be used, in particular chimerized, humanized or human forms of antibodies may be used. As the skilled person will know, alternative and/or equivalent names may be in use for certain antibodies mentioned in the present disclosure. Such alternative and/or equivalent names are interchangeable in the context of the present invention. For example, it is known that lambrolizumab is also known under the alternative and equivalent names MK-3475 and pembrolizumab.

[00139] It is contemplated that any of the immune checkpoint inhibitors that are known in the art to stimulate immune responses may be used. This includes inhibitors that directly or indirectly stimulate or enhance antigen-specific T-lymphocytes. These immune checkpoint inhibitors include, without limitation, agents targeting immune checkpoint proteins and pathways involving PD-L2, LAG3, BTLA, B7H4 and TIM3. For example, LAG3 inhibitors known in the art include soluble LAG3 (IMP321, or LAG3-Ig disclosed in W02009044273) as well as mouse or humanized antibodies blocking human LAG3 (e.g., IMP701 disclosed in W02008132601), or fully human antibodies blocking human LAG3 (such as disclosed in EP 2320940). Another example is provided by the use of blocking agents towards BTLA, including without limitation antibodies blocking human BTLA interaction with its ligand (such as 4C7 disclosed in WO2011014438). Yet another example is provided by the use of agents neutralizing B7H4 including without limitation antibodies to human B7H4 (disclosed in WO 2013025779, and in WO2013067492) or soluble recombinant forms of B7H4 (such as disclosed in US20120177645). Yet another example is provided by agents neutralizing B7-H3, including without limitation antibodies neutralizing human B7-H3 (e.g. MGA271 disclosed as BRCA84D and derivatives in US 20120294796). Yet another example is provided by agents targeting TIM3, including without limitation antibodies targeting human TIM3 (e.g. as disclosed in WO 2013006490 A2 or the anti-human TIM3, blocking antibody F38-2E2 disclosed by Jones et al. , 2008).

[00140] In addition, more than one immune checkpoint inhibitor (e.g., anti-PD- 1 antibody and anti-CTLA-4 antibody) may be used in combination with the p53 dendritic cell vaccine and CD 122/CD 132 treatment. For example, the p53 dendritic cell vaccine and CD 122/CD 132 treatment and immune checkpoint inhibitors (e.g., anti-CTLA4 antibody and/or anti-PD-l antibody) can be administered to enhance anti-tumor immunity.

A. PD-1 Axis Antagonists

[00141] T cell dysfunction or anergy occurs concurrently with an induced and sustained expression of the inhibitory receptor, programmed death 1 polypeptide (PD-l). Thus, therapeutic targeting of PD-l and other molecules which signal through interactions with PD- 1, such as programmed death ligand 1 (PD-L1) and programmed death ligand 2 (PD-L2) is provided herein. PD-L1 is overexpressed in many cancers and is often associated with poor prognosis (Okazaki T et al, 2007). Thus, inhibition of the PD-L1/PD-1 interaction in combination with the p53 dendritic cell vaccine and the CD122/CD132 treatment is provided herein such as to enhance CD8⁺ T cell-mediated killing of tumors.

[00142] Provided herein is a method for treating or delaying progression of cancer in an individual comprising administering to the individual an effective amount of a PD-l axis binding antagonist in combination with the p53 dendritic cell vaccine and the CD 122/CD 132 treatment. Also provided herein is a method of enhancing immune function in an individual in need thereof comprising administering to the individual an effective amount of a PD-l axis binding antagonist and the p53 dendritic cell vaccine and the CD122/CD132 treatment. [00143] For example, a PD-l axis binding antagonist includes a PD-l binding antagonist, a PDL1 binding antagonist and a PDL2 binding antagonist. Alternative names for "PD-l" include CD279 and SLEB2. Alternative names for "PDL1" include B7-H1, B7-4, CD274, and B7-H. Alternative names for "PDL2" include B7-DC, Btdc, and CD273. In some embodiments, PD-l, PDL1, and PDL2 are human PD-l, PDL1 and PDL2.

[00144] In some embodiments, the PD-l binding antagonist is a molecule that inhibits the binding of PD-l to its ligand binding partners. In a specific aspect, the PD-l ligand binding partners are PDL1 and/or PDL2. In another embodiment, a PDL1 binding antagonist is a molecule that inhibits the binding of PDL1 to its binding partners. In a specific aspect, PDL1 binding partners are PD-l and/or B7-1. In another embodiment, the PDL2 binding antagonist is a molecule that inhibits the binding of PDL2 to its binding partners. In a specific aspect, a PDL2 binding partner is PD-l. The antagonist may be an antibody, an antigen binding fragment thereof, an immunoadhesion, a fusion protein, or oligopeptide. Exemplary antibodies are described in U.S. Patent Nos. US8735553, US8354509, and US8008449, all incorporated herein by reference. Other PD- 1 axis antagonists for use in the methods provided herein are known in the art such as described in U.S. Patent Application No. US20140294898, US2014022021, and US20110008369, all incorporated herein by reference.

[00145] In some embodiments, the PD-l binding antagonist is an anti-PD-l antibody (e.g., a human antibody, a humanized antibody, or a chimeric antibody). In some embodiments, the anti-PD-l antibody is selected from the group consisting of nivolumab, pembrolizumab, and CT-011. In some embodiments, the PD-l binding antagonist is an immunoadhesin (e.g., an immunoadhesin comprising an extracellular or PD-l binding portion of PDL1 or PDL2 fused to a constant region (e.g., an Fc region of an immunoglobulin sequence). In some embodiments, the PD-l binding antagonist is AMP- 224. Nivolumab, also known as MDX-1106-04, MDX-1106, ONO-4538, BMS-936558, and OPDIVO^®, is an anti- PD-l antibody described in W02006/121168. Pembrolizumab, also known as MK-3475, Merck 3475, lambrolizumab, KEYTRUDA^®, and SCH-900475, is an anti-PD-l antibody described in W02009/114335. CT-011, also known as hBAT or hBAT-l, is an anti-PD-l antibody described in W02009/101611. AMP-224, also known as B7-DCIg, is a PDL2-Fc fusion soluble receptor described in W02010/027827 and WO2011/066342. Additional PD-l binding antagonists include Pidilizumab, also known as CT-011, MEDI0680, also known as AMP-514, and REGN2810. [00146] In some aspects, the immune checkpoint inhibitor is a PD-L1 antagonist such as Durvalumab, also known as MEDI4736, atezolizumab, also known as MPDL3280A, or avelumab, also known as MSB00010118C. In certain aspects, the immune checkpoint inhibitor is a PD-L2 antagonist such as rHIgMl2B7. In some aspects, the immune checkpoint inhibitor is a LAG-3 antagonist such as, but not limited to, IMP321, and BMS-986016. The immune checkpoint inhibitor may be an adenosine A2a receptor (A2aR) antagonist such as PBF-509.

[00147] In some aspects, the antibody described herein (such as an anti-PD-l antibody, an anti-PDLl antibody, or an anti-PDL2 antibody) further comprises a human or murine constant region. In a still further aspect, the human constant region is selected from the group consisting of IgGl, IgG2, IgG2, IgG3, IgG4. In a still further specific aspect, the human constant region is IgGl. In a still further aspect, the murine constant region is selected from the group consisting of IgGl, IgG2A, IgG2B, IgG3. In a still further specific aspect, the antibody has reduced or minimal effector function. In a still further specific aspect, the minimal effector function results from production in prokaryotic cells. In a still further specific aspect the minimal effector function results from an "effector-less Fc mutation" or aglycosylation.

[00148] Accordingly, an antibody used herein can be aglycosylated. Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine- X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxy amino acid, most commonly serine or threonine, although 5- hydroxyproline or 5 -hydroxy lysine may also be used. Removal of glycosylation sites form an antibody is conveniently accomplished by altering the amino acid sequence such that one of the above-described tripeptide sequences (for N-linked glycosylation sites) is removed. The alteration may be made by substitution of an asparagine, serine or threonine residue within the glycosylation site another amino acid residue (e.g., glycine, alanine or a conservative substitution).

[00149] The antibody or antigen binding fragment thereof, may be made using methods known in the art, for example, by a process comprising culturing a host cell containing nucleic acid encoding any of the previously described anti-PDLl, anti-PD-l, or anti-PDL2 antibodies or antigen-binding fragment in a form suitable for expression, under conditions suitable to produce such antibody or fragment, and recovering the antibody or fragment.

B. CTLA-4

[00150] Another immune checkpoint that can be targeted in the methods provided herein is the cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), also known as CD 152. The complete cDNA sequence of human CTLA-4 has the Genbank accession number L15006. CTLA-4 is found on the surface of T cells and acts as an“off’ switch when bound to CD80 or CD86 on the surface of antigen-presenting cells. CTLA4 is a member of the immunoglobulin superfamily that is expressed on the surface of Helper T cells and transmits an inhibitory signal to T cells. CTLA4 is similar to the T-cell co-stimulatory protein, CD28, and both molecules bind to CD80 and CD86, also called B7-1 and B7-2 respectively, on antigen-presenting cells. CTLA4 transmits an inhibitory signal to T cells, whereas CD28 transmits a stimulatory signal. Intracellular CTLA4 is also found in regulatory T cells and may be important to their function. T cell activation through the T cell receptor and CD28 leads to increased expression of CTLA-4, an inhibitory receptor for B7 molecules.

[00151] In some embodiments, the immune checkpoint inhibitor is an anti- CTLA-4 antibody (e.g., a human antibody, a humanized antibody, or a chimeric antibody), an antigen binding fragment thereof, an immunoadhesin, a fusion protein, or oligopeptide.

[00152] Anti-human-CTLA-4 antibodies (or VH and/or VL domains derived therefrom) suitable for use in the present methods can be generated using methods well known in the art. Alternatively, art recognized anti-CTLA-4 antibodies can be used. For example, the anti-CTLA-4 antibodies disclosed in: US 8,119,129, WO 01/14424, WO 98/42752; WO 00/37504 (CP675,206, also known as tremelimumab; formerly ticilimumab), U.S. Patent No. 6,207,156; Hurwitz et al, 1998; Camacho et al, 2004; and Mokyr et al, 1998 can be used in the methods disclosed herein. The teachings of each of the aforementioned publications are hereby incorporated by reference. Antibodies that compete with any of these art-recognized antibodies for binding to CTLA-4 also can be used. For example, a humanized CTLA-4 antibody is described in International Patent Application No. W02001014424, W02000037504, and U.S. Patent No. US8017114; all incorporated herein by reference. [00153] An exemplary anti-CTLA-4 antibody is ipilimumab (also known as 10D1, MDX- 010, MDX- 101, and Yervoy®) or antigen binding fragments and variants thereof (see, e.g. , WOO 1/14424). In other embodiments, the antibody comprises the heavy and light chain CDRs or VRs of ipilimumab. Accordingly, in one embodiment, the antibody comprises the CDR1, CDR2, and CDR3 domains of the VH region of ipilimumab, and the CDR1, CDR2 and CDR3 domains of the VL region of ipilimumab. In another embodiment, the antibody competes for binding with and/or binds to the same epitope on CTLA-4 as the above- mentioned antibodies. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with the above-mentioned antibodies (e.g., at least about 90%, 95%, or 99% variable region identity with ipilimumab).

[00154] Other molecules for modulating CTLA-4 include CTLA-4 ligands and receptors such as described in U.S. Patent Nos. US5844905, US5885796 and International Patent Application Nos. WO1995001994 and WO1998042752; all incorporated herein by reference, and immunoadhesions such as described in U.S. Patent No. US8329867, incorporated herein by reference.

C. Killer Immunoglobulin-like Receptor (KIR)

[00155] Another immune checkpoint inhibitor for use in the present invention is an anti-KIR antibody. Anti-human-KIR antibodies (or VH/VL domains derived therefrom) suitable for use in the invention can be generated using methods well known in the art.

[00156] Alternatively, art recognized anti-KIR antibodies can be used. The anti- KIR antibody can be cross-reactive with multiple inhibitory KIR receptors and potentiates the cytotoxicity of NK cells bearing one or more of these receptors. For example, the anti-KIR antibody may bind to each of KIR2D2DL1, KIR2DL2, and KIR2DL3, and potentiate NK cell activity by reducing, neutralizing and/or reversing inhibition of NK cell cytotoxicity mediated by any or all of these KIRs. In some aspects, the anti-KIR antibody does not bind KIR2DS4 and/or KIR2DS3. For example, monoclonal antibodies 1-7F9 (also known as IPH2101), 14F1, 1-6F1 and 1-6F5, described in WO 2006/003179, the teachings of which are hereby incorporated by reference, can be used. Antibodies that compete with any of these art- recognized antibodies for binding to KIR also can be used. Additional art-recognized anti-KIR antibodies which can be used include, for example, those disclosed in WO 2005/003168, WO 2005/009465, WO 2006/072625, WO 2006/072626, WO 2007/042573, WO 2008/084106, WO 2010/065939, WO 2012/071411 and WO/2012/160448. [00157] An exemplary anti-KIR antibody is lirilumab (also referred to as BMS- 986015 or IPH2102). In other embodiments, the anti-KIR antibody comprises the heavy and light chain complementarity determining regions (CDRs) or variable regions (VRs) of lirilumab. Accordingly, in one embodiment, the antibody comprises the CDR1, CDR2, and CDR3 domains of the heavy chain variable (VH) region of lirilumab, and the CDR1, CDR2 and CDR3 domains of the light chain variable (VL) region of lirilumab. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with lirilumab.

VII. Methods of Treatment

[00158] Provided herein are methods for treating, delaying progression of, or preventing cancer in an individual comprising administering to the individual an effective amount a p53 dendritic cell vaccine in combination with at least one CD122/CD132 agonist.

[00159] In some embodiments, the treatment results in a sustained response in the individual after cessation of the treatment. The methods described herein may find use in treating conditions where enhanced immunogenicity is desired such as increasing tumor immunogenicity for the treatment of cancer. Also provided herein are methods of enhancing immune function such as in an individual having cancer comprising administering to the individual an effective amount of a p53 dendritic cell vaccine and a preferential CD122/CD132 agonist. In some embodiments, the individual is a human.

[00160] Examples of cancers contemplated for treatment include lung cancer, head and neck cancer, breast cancer, pancreatic cancer, prostate cancer, renal cancer, bone cancer, testicular cancer, cervical cancer, gastrointestinal cancer, lymphomas, pre-neoplastic lesions in the lung, colon cancer, melanoma, and bladder cancer.

[00161] In some embodiments, the individual has cancer that is resistant (has been demonstrated to be resistant) to one or more anti-cancer therapies. In some embodiments, resistance to anti-cancer therapy includes recurrence of cancer or refractory cancer. Recurrence may refer to the reappearance of cancer, in the original site or a new site, after treatment. In some embodiments, resistance to anti-cancer therapy includes progression of the cancer during treatment with the anti-cancer therapy. In some embodiments, the cancer is at early stage or at late stage. [00162] The individual may have a cancer that expresses (has been shown to express e.g., in a diagnostic test) PD-L1 biomarker. In some embodiments, the patient's cancer expresses low PD-L1 biomarker. In some embodiments, the patient's cancer expresses high PD-L1 biomarker. In some embodiments the patient’s cancer expresses high levels of either normal or mutated p53. The PD-L1 and p53 biomarkers can be detected in the sample using a method selected from the group consisting of FACS, Western blot, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blotting, immunodetection methods, HPLC, surface plasmon resonance, optical spectroscopy, mass spectrometery, HPLC, qPCR, RT-qPCR, multiplex qPCR or RT-qPCR, RNA-seq, microarray analysis, SAGE, MassARRAY technique, and FISH, and combinations thereof.

[00163] The efficacy of any of the methods described herein (e.g., combination treatments including administering an effective amount of p53 dendritic cell vaccine and a preferential CD122/CD132 agonist) may be tested in various models known in the art, such as clinical or pre -clinical models. Suitable pre-clinical models are exemplified herein and further may include without limitation ID8 ovarian cancer, GEM models, B16F10 melanoma, RENCA renal cell cancer, CT26 colorectal cancer, MC38 colorectal cancer, D459 MethA sarcoma and Cloudman melanoma models of cancer.

[00164] In some embodiments of the methods of the present disclosure, the cancer has low levels of T cell infiltration. In some embodiments, the cancer has no detectable T cell infiltrate. In some embodiments, the cancer has low or undetectable levels of anti-p53 T cells. In some embodiments, the cancer is a non-immunogenic cancer (e.g., non-immunogenic colorectal cancer and/or ovarian cancer). Without being bound by theory, the combination treatment may increase T cell or anti-p53 T cell (e.g., CD4⁺ T cell, CD8⁺ T cell, memory T cell) priming, activation and/or proliferation relative to prior to the administration of the combination.

[00165] In some embodiments of the methods of the present disclosure, activated

CD4 and/or CD8 T cells in the individual are characterized by g-IEN producing CD4 and/or CD8 T cells and/or enhanced cytolytic activity relative to prior to the administration of the combination. g-IFN may be measured by any means known in the art, including, e.g., intracellular cytokine staining (ICS) involving cell fixation, permeabilization, and staining with an antibody against g-IEN. Cytolytic activity may be measured by any means known in the art, e.g., using a cell killing assay with mixed effector and target cells. [00166] The present disclosure is useful for any human cell that participates in an immune reaction either as a target for the immune system or as part of the immune system's response to the foreign target. The methods include ex vivo methods, in vivo methods, and various other methods that involve injection of polynucleotides or vectors into the host cell. The methods also include injection directly into the tumor or tumor bed as well as local or regional to the tumor.

A. Administration

[00167] The therapy provided herein comprises administration of an effective amount of p53 dendritic cell vaccine and a CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor (e.g., PD-l axis binding antagonist and/or CTLA-4 antibody). The combination therapy may be administered in any suitable manner known in the art. For example, of an immune checkpoint inhibitor (e.g., PD-l axis binding antagonist and/or CTLA-4 antibody), a p53 dendritic cell vaccine and CD122/CD132 agonist treatment may be administered sequentially (at different times) or concurrently (at the same time). In some embodiments, the one or more immune checkpoint inhibitors and the preferential CD122/CD132 agonist are in separate compositions as the p53 dendritic cell vaccine.

[00168] The p53 dendritic cell vaccine and the CD 122/CD 132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor may be administered by the same route of administration or by different routes of administration· In some embodiments, the preferential CD122/CD132 agonist and immune checkpoint inhibitor are administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. In some embodiments, p53 dendritic cell vaccine composition therapy is administered intradermally, intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. In some aspects, the administration is via continuous infusion, intratumoral injection, intravenous injection, intra-arterial injection, intra-peritoneal injection, intrapleural injection, or intra-thecal injection. An effective amount of the p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor may be administered for prevention or treatment of disease. The appropriate dosage of p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor therapy may be determined based on the type of disease to be treated, severity and course of the disease, the clinical condition of the individual, the individual's clinical history and response to the treatment, and the discretion of the attending physician. In some embodiments, p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor are synergistic, whereby an efficacious dose of p53 dendritic cell vaccine and the CD122/CD132 agonist treatment in combination with at least one immune checkpoint inhibitor has more than additive therapeutic benefit compared to each treatment as a single agent.

[00169] For example, the therapeutically effective amount of the p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor that is administered to a human will be in the range of about 0.001 to about 50 mg/kg of patient body weight whether by one or more administrations for the preferential CD122/CD132 agonist or immune checkpoint inhibitor antibody. In some embodiments, the antibody used is about 0.01 to about 45 mg/kg, about 0.01 to about 40 mg/kg, about 0.01 to about 35 mg/kg, about 0.01 to about 30 mg/kg, about 0.01 to about 25 mg/kg, about 0.01 to about 20 mg/kg, about 0.01 to about 15 mg/kg, about 0.01 to about 10 mg/kg, about 0.01 to about 5 mg/kg, or about 0.01 to about 1 mg/kg administered daily, for example. In some embodiments, the antibody is administered at 15 mg/kg. However, other dosage regimens may be useful. In one embodiment, an anti-PDLl antibody described herein is administered to a human at a dose of about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg or about 1400 mg on day 1 of 2l-day cycles. The dose may be administered as a single dose or as multiple doses (e.g., 2 or 3 doses or more doses), such as infusions or injections. The therapeutically effective amount of the preferential CD122/CD132 agonist, such as an IL2/anti-IL2 immune complex, ILl5/anti-ILl5 immune complex, an IL15/IL15 Receptor a-IgGl-Fc (ILl5/ILl5Ra-IgGl-Fc) immunocomplex, PEGylated IL2, PEGylated IL15, IL2 muteins, and/or IL15 muteins) includes but is not limited to administration in doses ranging between 5-100 ug/kg given either subcutaneously (SQ) or intravenously (IV) at intervals ranging from weekly to every 2-4 weeks per treatment cycle. More than one treatment cycle may be given. [00170] Treatment with the p53 dendritic cell vaccine component comprises administering at least 1, 2, 3, 4, 5 or 6 doses of dendritic cells per treatment cycle at 1 x 10⁶ to 1 x 10⁷ cells per dose, including about 1 x 10⁶, 2 x 10⁶, 3 x 10⁶, 4 x 10⁶, 5 x 10⁶, 6 x 10⁶, 7 x 10⁶, 8 x 10⁶, 9 x 10⁶, and 1 x 10⁷ cells per dose. These doses can be administered at 1, 2, 3, 4, 5, or 6 day intervals or 1, 2, 3, or 4 week intervals. Multiple cycles of p53 dendritic cell vaccinations can be given. The progress of therapy is easily monitored by conventional techniques.

[00171] Treatment regimens may vary as well, and often depend on tumor type, tumor location, disease progression, and health and age of the patient. Obviously, certain types of tumors will require more aggressive treatment, while at the same time, certain patients cannot tolerate more taxing protocols. The clinician will be best suited to make such decisions based on the known efficacy and toxicity (if any) of the therapeutic formulations.

[00172] In certain embodiments, the tumor being treated may not, at least initially, be resectable. Administration of the p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor may increase the resectability of the tumor due to shrinkage at the margins or by elimination of certain particularly invasive portions. Following treatments, resection may be possible. Additional treatments subsequent to resection will serve to eliminate microscopic residual disease.

[00173] The treatments may include various "unit doses." Unit dose is defined as containing a predetermined-quantity of the therapeutic composition. The quantity to be administered, and the particular route and formulation, are within the skill of those in the clinical arts. A unit dose need not be administered as a single injection but may comprise continuous infusion over a set period of time. Unit dose of the present invention may conveniently be described in terms of plaque forming units (pfu) for a viral construct. Unit doses range from 10³, 10⁴, 10⁵, 10⁶, 10⁷, 10^s, 10⁹, 10¹⁰, 10¹¹, 10¹², 10¹³ pfu and higher. Alternatively, depending on the kind of virus and the titer attainable, one will deliver 1 to 100, 10 to 50, 100-1000, or up to about 1 x 10⁴, 1 x 10⁵, 1 x 10⁶, 1 x 10⁷, 1 x 10⁸ , 1 x 10⁹, 1 x 10¹⁰, 1 x 10¹¹, 1 x 10¹², 1 x 10¹³, 1 x 10¹⁴, or 1 x 10¹⁵ or higher infectious viral particles (vp) to the patient or to the patient's cells. B. Injectable Compositions and Formulations

[00174] One method for the delivery of the p53 dendritic cell vaccine component in the present disclosure is via intradermal injection at a non-tumor site. However, the pharmaceutical compositions disclosed herein may alternatively be administered parenterally, intravenously, intradermally, intramuscularly, transdermally or even intraperitoneally as described in U.S. Patent 5,543,158, U.S. Patent 5,641,515 and U.S. Patent 5,399,363, all incorporated herein by reference.

[00175] Injection of nucleic acid constructs as additional treatments may be delivered by syringe or any other method used for injection of a solution, as long as the expression construct can pass through the particular gauge of needle required for injection. A novel needleless injection system has been described (U.S. Patent 5,846,233) having a nozzle defining an ampule chamber for holding the solution and an energy device for pushing the solution out of the nozzle to the site of delivery. A syringe system has also been described for use in gene therapy that permits multiple injections of predetermined quantities of a solution precisely at any depth (U.S. Patent 5,846,225).

[00176] Solutions of the active compounds as free base or pharmacologically acceptable salts may be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (U.S. Patent 5,466,468). In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and/or vegetable oils. Proper fluidity may be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[00177] For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous, intratumoral and intraperitoneal administration· In this connection, sterile aqueous media that can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage may be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, "Remington's Pharmaceutical Sciences" 22md Edition). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.

[00178] Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vaccuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[00179] The compositions disclosed herein may be formulated in a neutral or salt form. Pharmaceutically-acceptable salts, include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like.

C. Additional Anti-Cancer Therapies

[00180] In order to increase the effectiveness of the p53 dendritic cell vaccine and preferential CD122/CD132 agonist treatment provided herein and, in some aspects, the at least one immune checkpoint inhibitor, they can be combined with at least one additional agent effective in the treatment of cancer. More generally, these other therapies or compositions would be provided in a combined manner or amount effective to remove, kill or inhibit proliferation of the tumor cells. This process may involve applying therapies, the agent(s) or multiple factor(s) at the same time. This may be achieved by applying the treatment modalities at the same time or sequentially to enhance anti-tumor immune responses and therapeutic efficacy.

[00181] The at least one additional anticancer therapy may be, without limitation, a surgical therapy, chemotherapy (e.g., administration of a protein kinase inhibitor or a EGFR-targeted therapy), radiation therapy, cryotherapy, hyperthermia treatment, phototherapy, radioablation therapy, hormonal therapy, immunotherapy, small molecule therapy, receptor kinase inhibitor therapy, anti-angiogenic therapy, cytokine therapy or a biological therapies such as monoclonal antibodies, siRNA, miRNA, antisense oligonucleotides, ribozymes or gene therapy. Without limitation the biological therapy may be an oncolytic viral therapy, gene therapy, such as tumor suppressor gene therapy, a cell death protein gene therapy, a cell cycle regulator gene therapy, a cytokine gene therapy, a toxin gene therapy, an immunogene therapy, a suicide gene therapy, a prodrug gene therapy, an anti- cellular proliferation gene therapy, an enzyme gene therapy, an anti- angiogenic factor gene therapy or another type of tumor vaccine therapy.

[00182] The additional therapy may precede or follow the p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor treatment by intervals ranging from minutes to weeks. In embodiments where the other agent is applied separately to the cell, one would generally ensure that a significant period of time did not expire between the time of each delivery, such that the other agent would still be able to exert an advantageously combined effect on the cell. In such instances, it is contemplated that one may contact the cell with both modalities within about 12-24 hours of each other and, more preferably, within about 6-12 hours of each other. In some situations, it may be desirable to extend the time period for treatment significantly, however, where several days (e.g., 2, 3, 4, 5, 6 or 7) to several weeks (e.g., 1, 2, 3, 4, 5, 6, 7 or 8) lapse between the respective administrations·

[00183] Various combinations may be employed, p53 dendritic cell vaccine and the CD122/CD132 agonist treatment, in some embodiments, is "A" and the secondary agent, i.e. immune checkpoint inhibitor, is "B":

A/B/A B/A/B B/B/A A/A/B A/B/B B/A/A A/B/B/B B/A/B/B

B/B/B/A B/B/A/B A/A/B/B A/B/A/B A/B/B/A B/B/A/A

B/A/B/A B/A/A/B A/A/A/B B/A/A/A A/B/A/A A/A/B/A

1. Chemotherapy

[00184] Cancer therapies in general also include a variety of combination therapies with both chemical and radiation based treatments. Combination chemotherapies include, for example, cisplatin (CDDP), carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide (VP16), tamoxifen, raloxifene, estrogen receptor binding agents, taxol, gemcitabien, navelbine, famesyl-protein transferase inhibitors, transplatinum, 5-fluorouracil, vincristine, vinblastine and methotrexate, Temazolomide (an aqueous form of DTIC), or any analog or derivative variant of the foregoing. The combination of chemotherapy with biological therapy is known as biochemotherapy. The chemotherapy may also be administered at low, continuous doses which is known as metronomic chemotherapy.

[00185] Yet further combination chemotherapies include, for example, alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammall and calicheamicin omegall; dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromophores, aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino- doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxy doxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, pteropterin, trimetrexate; purine analogs such as fludarabine, 6- mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK polysaccharide complex; razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2”-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; taxoids, e.g., paclitaxel and docetaxel gemcitabine; 6-thioguanine; mercaptopurine; platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP- 16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-ll); topoisomerase inhibitor RFS 2000; difluorometlhylornithine (DMFO); retinoids such as retinoic acid; capecitabine; carboplatin, procarbazine, plicomycin, gemcitabien, navelbine, farnesyl-protein transferase inhibitors, transplatinum; and pharmaceutically acceptable salts, acids or derivatives of any of the above. In certain embodiments, the compositions provided herein may be used in combination with histone deacetylase inhibitors. In certain embodiments, the compositions provided herein may be used in combination with gefitinib. In other embodiments, the present embodiments may be practiced in combination with Gleevec (e.g., from about 400 to about 800 mg/day of Gleevec may be administered to a patient). In certain embodiments, one or more chemotherapeutic may be used in combination with the compositions provided herein.

2. Radiotherapy

[00186] Other factors that cause DNA damage and have been used extensively include what are commonly known as y-rays, X-rays, and/or the directed delivery of radioisotopes to tumor cells. Other forms of DNA damaging factors are also known such as microwaves and UV-irradiation. It is most likely that all of these factors effect a broad range of damage on DNA, on the precursors of DNA, on the replication and repair of DNA, and on the assembly and maintenance of chromosomes. Dosage ranges for X-rays range from daily doses of 50 to 200 roentgens for prolonged periods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens. Dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the uptake by the neoplastic cells.

3. Immunotherapy

[00187] Immunotherapeutics, generally, rely on the use of immune effector cells and molecules to target and destroy cancer cells. The immune effector may be, for example, an antibody specific for some marker on the surface of a tumor cell. The antibody alone may serve as an effector of therapy or it may recruit other cells to actually effect cell killing. The antibody also may be conjugated to a drug or toxin (chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussis toxin, etc.) and serve merely as a targeting agent. Alternatively, the effector may be a lymphocyte carrying a surface molecule that interacts, either directly or indirectly, with a tumor cell target. Various effector cells include cytotoxic T cells and NK cells as well as genetically engineered variants of these cell types modified to express chimeric antigen receptors.

[00188] It will be appreciated by those skilled in the art of cancer immunotherapy that other complementary immune therapies may be added to the regimens described above to further enhance their efficacy including but not limited to GM-CSF to increase the number of myeloid derived innate immune system cells, low dose cyclophosphamide or PI3K inhibitors (e.g., PI3K delta inhibitors) to eliminate T regulatory cells that inhibit innate and adaptive immunity and 5FU (e.g., capecitabine), PI3K inhibitors or histone deacetylase inhibitors to remove inhibitory myeloid derived suppressor cells. For example, PI3K inhibitors include, but are not limited to, LY294002, Perifosine, BKM120, Duvelisib, PX-866, BAY 80-6946, BEZ235, SF1126, GDC-0941, XL147, XL765, Palomid 529, GSK1059615, PWT33597, IC87114, TG100-15, CAL263, PI- 103, GNE-477, CUDC-907, and AEZS-136. In some aspects, the PI3K inhibitor is a PI3K delta inhibitor such as, but not limited to, Idelalisib, RP6530, TGR1202, and RP6503. Additional PI3K inhibitors are disclosed in U.S. Patent Application Nos. US20150291595, US20110190319, and International Patent Application Nos. WO2012146667, WO2014164942, WO2012062748, and WO2015082376. The immunotherapy may also comprise the administration of an interleukin such as IL-2, or an interferon such as INFa.

[00189] Examples of immunotherapies that can be combined with the p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor are immune adjuvants (e.g., Mycobacterium bo vis, Plasmodium falciparum, dinitrochlorobenzene and aromatic compounds) (U.S. Patent 5,801,005 ; U.S. Patent 5,739,169 ; Hui and Hashimoto, 1998; Christodoulides et ah, 1998), cytokine therapy (e.g., interferons a, b and g; interleukins (IL-l, IL-2), GM-CSF and TNF) (Bukowski et ah, 1998; Davidson et ah, 1998; Hellstrand et ah, 1998) gene therapy (e.g., TNF, IL-l, IL-2, IL-12, p53) (Qin et ah, 1998; Austin-Ward and Villaseca, 1998; U.S. Patent 5,830,880 and U.S. Patent 5,846,945 ) and monoclonal antibodies (e.g., anti-ganglioside GM2, anti-HER-2, anti-pl85) (Pietras et ah, 1998; Hanibuchi et ah, 1998; U.S. Patent 5,824,311 ). Herceptin (trastuzumab) is a chimeric (mouse-human) monoclonal antibody that blocks the HER2-neu receptor. It possesses anti-tumor activity and has been approved for use in the treatment of malignant tumors (Dillman, 1999). Combination therapy of cancer with herceptin and chemotherapy has been shown to be more effective than the individual therapies. Thus, it is contemplated that one or more anti-cancer therapies may be employed with the Ad-mda7 therapy described herein.

[00190] Additional immunotherapies that may be combined with p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor include a co-stimulatory receptor agonist, a stimulator of innate immune cells, or an activator of innate immunity. The co-stimulatory receptor agonist may be an anti-OX40 antibody (e.g., MEDI6469, MEDI6383, MEDI0562, and MOXR0916), anti- GITR antibody (e.g., TRX518, and MK-4166), anti-CDl37 antibody (e.g., Urelumab, and PF- 05082566), anti-CD40 antibody (e.g., CP-870,893, and Chi Lob 7/4), or an anti-CD27 antibody (e.g., Varlilumab, also known as CDX-1127). The stimulators of innate immune cells include, but are not limited to, a KIR monoclonal antibody (e.g., lirilumab), an inhibitor of a cytotoxicity-inhibiting receptor (e.g., NKG2A, also known as KLRC and as CD94, such as the monoclonal antibody monalizumab, and anti-CD96,also known as TACTILE), and a toll like receptor (TLR) agonist. The TLR agonist may be BCG, a TLR7 agonist (e.g., polyOICLC, and imiquimod), a TLR8 agonist (e.g., resiquimod), or a TLR9 agonist (e.g., CPG 7909). The activators of innate immune cells, such as natural killer (NK) cells, macrophages, and dendritic cells, include IDO inhibitors, TGF inhibitor, IL-10 inhibitor. An exemplary activator of innate immunity is Indoximod. In some aspects, the immunotherapy is a stimulator of interferon genes (STING) agonist (Corrales et al., 2015).

[00191] Other immunotherapies contemplated for use in methods of the present disclosure include those described by Tchekmedyian et al, 2015, incorporated herein by reference. The immunotherapy may comprise suppression of T regulatory cells (Tregs), myeloid derived suppressor cells (MDSCs) and cancer associated fibroblasts (CAFs). In some embodiments, the immunotherapy is another tumor vaccine (e.g., whole tumor cell vaccines, peptides, and recombinant tumor associated antigen vaccines), or adoptive cellular therapies (ACT) (e.g., T cells, natural killer cells, TILs, and LAK cells). The T cells may be engineered with chimeric antigen receptors (CARs) or T cell receptors (TCRs) to specific tumor antigens. As used herein, a chimeric antigen receptor (or CAR) may refer to any engineered receptor specific for an antigen of interest that, when expressed in a T cell, confers the specificity of the CAR onto the T cell. Once created using standard molecular techniques, a T cell expressing a chimeric antigen receptor may be introduced into a patient, as with a technique such as adoptive cell transfer. In some aspects, the T cells are activated CD4 and/or CD8 T cells in the individual which are characterized by g-IFN producing CD4 and/or CD 8 T cells and/or enhanced cytolytic activity relative to prior to the administration of the combination. The CD4 and/or CD8 T cells may exhibit increased release of cytokines selected from the group consisting of IFN- g, TNF-aand interleukins. The CD4 and/or CD8 T cells can be effector memory T cells. In certain embodiments, the CD4 and/or CD8 effector memory T cells are characterized by having the expression of CD44^hlgh CD62L^low.

[00192] In certain aspects, two or more immunotherapies may be combined with p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor including additional immune checkpoint inhibitors in combination with agonists of T-cell costimulatory receptors, or in combination with TIL ACT. Other combinations include T-cell checkpoint blockade plus costimulatory receptor agonists, T-cell checkpoint blockade to improve innate immune cell function, checkpoint blockade plus IDO inhibition, or checkpoint blockade plus adoptive T-cell transfer. In certain aspects, immunotherapy includes a combination of an anti-PD-Ll immune checkpoint inhibitor (e.g., Avelumab), a 4-1BB (CD-137) agonist (e.g. Utomilumab), and an 0X40 (TNFRS4) agonist. The immunotherapy may be combined with histone deacetylase (HD AC) inhibitors such as 5-azacytidine and entinostat.

[00193] The immunotherapy may be another cancer vaccine comprising one or more cancer antigens, in particular a protein or an immunogenic fragment thereof, DNA or RNA encoding said cancer antigen, in particular a protein or an immunogenic fragment thereof, cancer cell lysates, and/or protein preparations from tumor cells. As used herein, a cancer antigen is an antigenic substance present in cancer cells. In principle, any protein produced in a cancer cell that has an abnormal structure due to mutation can act as a cancer antigen. In principle, cancer antigens can be products of mutated Oncogenes and tumor suppressor genes, products of other mutated genes, overexpressed or aberrantly expressed cellular proteins, cancer antigens produced by oncogenic viruses, oncofetal antigens, altered cell surface glycolipids and glycoproteins, or cell type-specific differentiation antigens. Examples of cancer antigens include the abnormal products of ras and p53 genes. Other examples include tissue differentiation antigens, mutant protein antigens, oncogenic viral antigens, cancer-testis antigens and vascular or stromal specific antigens. Tissue differentiation antigens are those that are specific to a certain type of tissue. Mutant protein antigens are likely to be much more specific to cancer cells because normal cells shouldn't contain these proteins. Normal cells will display the normal protein antigen on their MHC molecules, whereas cancer cells will display the mutant version. Some viral proteins are implicated in forming cancer, and some viral antigens are also cancer antigens. Cancer-testis antigens are antigens expressed primarily in the germ cells of the testes, but also in fetal ovaries and the trophoblast. Some cancer cells aberrantly express these proteins and therefore present these antigens, allowing attack by T- cells specific to these antigens. Exemplary antigens of this type are CTAG1 B and MAGEA1 as well as Rindopepimut, a l4-mer intradermal injectable peptide vaccine targeted against epidermal growth factor receptor (EGFR) vlll variant. Rindopepimut is particularly suitable for treating glioblastoma when used in combination with an inhibitor of the CD95/CD95L signaling system as described herein. Also, proteins that are normally produced in very low quantities, but whose production is dramatically increased in cancer cells, may trigger an immune response. An example of such a protein is the enzyme tyrosinase, which is required for melanin production. Normally tyrosinase is produced in minute quantities but its levels are very much elevated in melanoma cells. Oncofetal antigens are another important class of cancer antigens. Examples are alphafetoprotein (AFP) and carcinoembryonic antigen (CEA). These proteins are normally produced in the early stages of embryonic development and disappear by the time the immune system is fully developed. Thus self-tolerance does not develop against these antigens. Abnormal proteins are also produced by cells infected with oncoviruses, e.g. EBV and HPV. Cells infected by these viruses contain latent viral DNA which is transcribed and the resulting protein produces an immune response. A cancer vaccine may include a peptide cancer vaccine, which in some embodiments is a personalized peptide vaccine. In some embodiments, the peptide cancer vaccine is a multivalent long peptide vaccine, a multi -peptide vaccine, a peptide cocktail vaccine, a hybrid peptide vaccine, or a peptide-pulsed dendritic cell vaccine

[00194] The immunotherapy may be an antibody, such as part of a polyclonal antibody preparation, or may be a monoclonal antibody. The antibody may be a humanized antibody, a chimeric antibody, an antibody fragment, a bispecific antibody or a single chain antibody. An antibody as disclosed herein includes an antibody fragment, such as, but not limited to, Fab, Fab' and F(ab')2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdfv) and fragments including either a VL or VH domain. In some aspects, the antibody or fragment thereof specifically binds epidermal growth factor receptor (EGFR1, Erb-Bl), HER2/neu (Erb-B2), CD20, Vascular endothelial growth factor (VEGF), insulin- like growth factor receptor (IGF-1R), TRAIL-receptor, epithelial cell adhesion molecule, carcino-embryonic antigen, Prostate-specific membrane antigen, Mucin- 1, CD30, CD33, or CD40.

[00195] Examples of monoclonal antibodies that may be used in combination with the compositions provided herein include, without limitation, trastuzumab (anti- HER2/neu antibody); Pertuzumab (anti-HER2 mAb); cetuximab (chimeric monoclonal antibody to epidermal growth factor receptor EGFR); panitumumab (anti-EGFR antibody); nimotuzumab (anti-EGFR antibody); Zalutumumab (anti-EGFR mAb); Necitumumab (anti- EGFR mAb); MDX-210 (humanized anti-HER-2 bispecific antibody); MDX-210 (humanized anti-HER-2 bispecific antibody); MDX-447 (humanized anti-EGF receptor bispecific antibody); Rituximab (chimeric murine/human anti-CD20 mAb); Obinutuzumab (anti-CD20 mAb); Ofatumumab (anti-CD20 mAb); Tositumumab-Il3l (anti-CD20 mAb); Ibritumomab tiuxetan (anti-CD20 mAb); Bevacizumab (anti-VEGF mAb); Ramucirumab (anti-VEGFR2 mAb); Ranibizumab (anti-VEGF mAb); Aflibercept (extracellular domains of VEGFR1 and VEGFR2 fused to IgGl Fc); AMG386 (angiopoietin- 1 and -2 binding peptide fused to IgGl Fc); Dalotuzumab (anti-IGF-lR mAb); Gemtuzumab ozogamicin (anti-CD33 mAb); Alemtuzumab (anti-Campath-l/CD52 mAb); Brentuximab vedotin (anti-CD30 mAb); Catumaxomab (bispecific mAh that targets epithelial cell adhesion molecule and CD3); Naptumomab (anti-5T4 mAb); Girentuximab (anti-Carbonic anhydrase ix); or Farletuzumab (anti-folate receptor). Other examples include antibodies such as Panorex™ (17-1A) (murine monoclonal antibody); Panorex (@ (17-1A) (chimeric murine monoclonal antibody); BEC2 (ami-idiotypic mAb, mimics the GD epitope) (with BCG); Oncolym (Lym-l monoclonal antibody); SMART M195 Ab, humanized 13' 1 LYM-l (Oncolym), Ovarex (B43.13, anti- idiotypic mouse mAb); 3622W94 mAh that binds to EGP40 (17-1 A) pancarcinoma antigen on adenocarcinomas; Zenapax (SMART Anti-Tac (IL-2 receptor); SMART M195 Ab, humanized Ab, humanized); NovoMAb-G2 (pancarcinoma specific Ab); TNT (chimeric mAh to histone antigens); TNT (chimeric mAh to histone antigens); Gliomab-H (Monoclonals— Humanized Abs); GNI-250 Mab; EMD-72000 (chimeric-EGF antagonist); LymphoCide (humanized IL.L.2 antibody); and MDX-260 bispecific, targets GD-2, ANA Ab, SMART IDIO Ab, SMART ABL 364 Ab or ImmuRAIT-CEA. Examples of antibodies include those disclosed in U.S. Pat. No. 5,736,167, U.S. Pat. No. 7,060,808, and U.S. Pat. No. 5,821,337.

[00196] Further examples of antibodies include Zanulimumab (anti-CD4 mAb), Keliximab (anti-CD4 mAb); Ipilimumab (MDX-101; anti-CTLA-4 mAb); Tremilimumab (anti-CTLA-4 mAb); (Daclizumab (anti-CD25/IL-2R mAb); Basiliximab (anti-CD25/IL-2R mAb); MDX-1106 (anti-PDl mAb); antibody to GITR; GC1008 (anti-TGF-b antibody); metelimumab/CAT-l92 (anti-TGF-b antibody); lerdelimumab/CAT-l52 (anti-TGF-b antibody); ID11 (anti-TGF-b antibody); Denosumab (anti-RANKL mAb); BMS-663513 (humanized anti-4- 1BB mAb); SGN-40 (humanized anti-CD40 mAb); CP870,893 (human anti-CD40 mAb); Infliximab (chimeric anti-TNF mAb; Adalimumab (human anti-TNF mAb); Certolizumab (humanized Fab anti-TNF); Golimumab (anti-TNF); Etanercept (Extracellular domain of TNFR fused to IgGl Fc); Belatacept (Extracellular domain of CTLA-4 fused to Fc); Abatacept (Extracellular domain of CTLA-4 fused to Fc); Belimumab (anti-B Lymphocyte stimulator); Muromonab-CD3 (anti-CD3 mAb); Otelixizumab (anti-CD3 mAb); Teplizumab (anti-CD3 mAb); Tocilizumab (anti-IL6R mAb); REGN88 (anti-IL6R mAb); Ustekinumab (anti-IL- 12/23 mAb); Briakinumab (anti-IL- 12/23 mAb); Natalizumab (anti-a4 integrin); Vedolizumab (anti-a4 b7 integrin mAb); T1 h (anti-CD6 mAb); Epratuzumab (anti-CD22 mAb); Efalizumab (anti-CDlla mAb); and Atacicept (extracellular domain of transmembrane activator and calcium-modulating ligand interactor fused with Fc). a. Passive Immunotherapy

[00197] A number of different approaches for passive immunotherapy of cancer exist. They may be broadly categorized into the following: injection of antibodies alone; injection of antibodies coupled to toxins or chemotherapeutic agents; injection of antibodies coupled to radioactive isotopes; injection of anti-idiotype antibodies; and finally, purging of tumor cells in bone marrow.

[00198] Preferably, human monoclonal antibodies are employed in passive immunotherapy, as they produce few or no side effects in the patient. Human monoclonal antibodies to ganglioside antigens have been administered intralesionally to patients suffering from cutaneous recurrent melanoma (Irie & Morton, 1986). Regression was observed in six out of ten patients, following, daily or weekly, intralesional injections. In another study, moderate success was achieved from intralesional injections of two human monoclonal antibodies (Irie et al„ 1989).

[00199] It may be favorable to administer more than one monoclonal antibody directed against two different antigens or even antibodies with multiple antigen specificity. Treatment protocols also may include administration of lymphokines or other immune enhancers as described by Bajorin et al. (1988). The development of human monoclonal antibodies is described in further detail elsewhere in the specification. b. Active Immunotherapy

[00200] In active immunotherapy, an antigenic peptide, polypeptide or protein, or an autologous or allogenic tumor cell composition or“vaccine” is administered, generally with a distinct bacterial adjuvant (Ravindranath & Morton, 1991; Morton & Ravindranath, 1996; Morton et al., 1992; Mitchell et al., 1990; Mitchell et al., 1993). In melanoma immunotherapy, those patients who elicit high IgM response often survive better than those who elicit no or low IgM antibodies (Morton et al., 1992). IgM antibodies are often transient antibodies and the exception to the rule appears to be anti-ganglioside or anticarbohydrate antibodies. c. Adoptive Immunotherapy

[00201] In adoptive immunotherapy, the patient's circulating lymphocytes, or tumor infiltrated lymphocytes, are isolated in vitro, activated by lymphokines such as IL-2 or transduced with genes for tumor necrosis, and readministered (Rosenberg et al., 1988; 1989). To achieve this, one would administer to an animal, or human patient, an immunologically effective amount of activated lymphocytes in combination with an adjuvant-incorporated antigenic peptide composition as described herein. The activated lymphocytes will most preferably be the patient's own cells that were earlier isolated from a blood or tumor sample and activated (or "expanded") in vitro. This form of immunotherapy has produced several cases of regression of melanoma and renal carcinoma, but the percentage of responders were few compared to those who did not respond. More recently, higher response rates have been observed when such adoptive immune cellular therapies have incorporated genetically engineered T cells that express chimeric antigen receptors (CAR) termed CAR T cell therapy. Similarly, natural killer cells both autologous and allogenic have been isolated, expanded and genetically modified to express receptors or ligands to facilitate their binding and killing of tumor cells.

4. Other Agents

[00202] It is contemplated that other agents may be used in combination with the compositions provided herein to improve the therapeutic efficacy of treatment. These additional agents include immunomodulatory agents, agents that affect the upregulation of cell surface receptors and GAP junctions, cytostatic and differentiation agents, inhibitors of cell adhesion, or agents that increase the sensitivity of the hyperproliferative cells to apoptotic inducers. Immunomodulatory agents include tumor necrosis factor; interferon alpha, beta, and gamma; IL-2 and other cytokines; F42K and other cytokine analogs; or MIP-l, MIP-lbeta, MCP-l, RANTES, and other chemokines. It is further contemplated that the upregulation of cell surface receptors or their ligands such as Fas / Fas ligand, DR4 or DR5 / TRAIL would potentiate the apoptotic inducing abilities of the compositions provided herein by establishment of an autocrine or paracrine effect on hyperproliferative cells. Increases intercellular signaling by elevating the number of GAP junctions would increase the anti-hyperproliferative effects on the neighboring hyperproliferative cell population. In other embodiments, cytostatic or differentiation agents can be used in combination with the compositions provided herein to improve the anti-hyerproliferative efficacy of the treatments. Inhibitors of cell adhesion are contemplated to improve the efficacy of the present invention. Examples of cell adhesion inhibitors are focal adhesion kinase (FAKs) inhibitors and Lovastatin. It is further contemplated that other agents that increase the sensitivity of a hyperproliferative cell to apoptosis, such as the antibody c225, could be used in combination with the compositions provided herein to improve the treatment efficacy.

[00203] In further embodiments, the other agents may be one or more oncolytic viruses, such as an oncolytic vims engineered to express a tumor suppressor gene like p53 and/or IL24, and/or a cytokine. Examples of oncolytic viruses include single or double stranded DNA viruses, RNA viruses, adenoviruses, adeno-associated viruses, retroviruses, lentiviruses, herpes viruses, pox viruses, vaccinia viruses, vesicular stomatitis viruses, polio viruses, Newcastle’s Disease viruses, Epstein-Barr viruses, influenza viruses and reoviruses, myxoma viruses, maraba viruses, rhabdoviruses, enadenotucirev or coxsackie viruses. In a particular embodiment, the other agent is talimogene laherparepvec (T-VEC) which is an oncolytic herpes simplex vims genetically engineered to express GM-CSF. Talimogene laherparepvec, HSV-l [strain JS1] ICP34.5-/ICP47-/hGM-CSF, (previously known as OncoVEX^{GM CSF}), is an intratumorally delivered oncolytic immunotherapy comprising an immune-enhanced HSV-l that selectively replicates in solid tumors. (Lui et al., Gene Therapy, 10:292-303, 2003; U.S. Patent No. 7,223,593 and U.S. Patent No. 7,537,924; incorporated herein by reference). In October 2015, the US FDA approved T-VEC, under the brand name IMLYGIC™, for the treatment of melanoma in patients with inoperable tumors. The characteristics and methods of administration of T-VEC are described in, for example, the IMLYGIC™ package insert (Amgen, 2015) and U.S. Patent Publication No. US2015/0202290; both incorporated herein by reference. For example, talimogene laherparepvec is typically administered by intratumoral injection into injectable cutaneous, subcutaneous, and nodal tumors at a dose of up to 4.0 ml of 10⁶ plaque forming unit/mL (PFU/mL) at day 1 of week 1 followed by a dose of up to 4.0 ml of 10⁸ PFU/mL at day 1 of week 4, and every 2 weeks (± 3 days) thereafter. The recommended volume of talimogene laherparepvec to be injected into the tumor(s) is dependent on the size of the tumor(s) and should be determined according to the injection volume guideline. While T-VEC has demonstrated clinical activity in melanoma patients, many cancer patients either do not respond or cease responding to T-VEC treatment. In one embodiment, p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor may be administered after, during or before T-VEC therapy, such as to reverse treatment resistance. Exemplary oncolytic viruses include, but are not limited to, Ad5-yCD/mutTKSR39rep-hILl2, Cavatak™, CG0070, DNX-2401, G207, HF10, IMLYGIC™, JX-594, MG1-MA3, MV-NIS, OBP-301, Reolysin®, Toca 511, Oncorine, and RIGVIR. Other exemplary oncolytic viruses are described, for example, in International Patent Publication Nos. WO2015/027163, WO2014/138314, W02014/047350, and WO2016/009017; all incorporated herein by reference.

[00204] In certain embodiments, hormonal therapy may also be used in conjunction with the present embodiments or in combination with any other cancer therapy previously described. The use of hormones may be employed in the treatment of certain cancers such as breast, prostate, ovarian, or cervical cancer to lower the level or block the effects of certain hormones such as testosterone or estrogen. This treatment is often used in combination with at least one other cancer therapy as a treatment option or to reduce the risk of metastases.

[00205] In some aspects, the at least one additional anticancer treatment is an inhibitor (e.g., small molecule inhibitor) of HDM2 (also known as MDM2) and/or HDM4, such as to block p53 activity. In specific aspects, the small molecule inhibitor of HDM2 is HDM201, cis-imidazolines (e.g., Nutlins), benzodiazepines (BDPs), and spiro-oxindoles. Other exemplary HDM2 and/or HDM4 inhibitors for use in the present methods are described in, for example, Carry et al, 2013; Patel and Player, 2008; U.S. Patent No. 8,846,657; International Patent Publication No. WO2014123882; U.S. Patent No. 9,073,898; and International Patent Publication No. WO2014115080; all incorporated herein by reference. [00206] In some aspects, the additional anti-cancer agent is a protein kinase inhibitor or a monoclonal antibody that inhibits receptors involved in protein kinase or growth factor signaling pathways such as an EGFR, VEGFR, AKT, Erbl, Erb2, ErbB, Syk, Bcr-Abl, JAK, Src, GSK-3, PI3K, Ras, Raf, MAPK, MAPKK, mTOR, c-Kit, eph receptor or BRAF inhibitors. Nonlimiting examples of protein kinase or growth factor signaling pathways inhibitors include Afatinib, Axitinib, Bevacizumab, Bosutinib, Cetuximab, Crizotinib, Dasatinib, Erlotinib, Fostamatinib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Mubritinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Saracatinib, Sorafenib, Sunitinib, Trastuzumab, Vandetanib, AP23451, Vemurafenib, MK-2206, GSK690693, A-443654, VQD-002, Miltefosine, Perifosine, CAL101, PX-866, LY294002, rapamycin, temsirolimus, everolimus, ridaforolimus, Alvocidib, Genistein, Selumetinib, AZD- 6244, Vatalanib, P1446A-05, AG-024322, ZD1839, P276-00, GW572016 or a mixture thereof. In certain aspects, the additional anti-cancer agent is a tyrosine kinase inhibitor, such as a Bruton’s tyrosine kinase (BTK) inhibitor. In some aspects, a small molecule BTK inhibitor as employed herein refers to a chemically synthesized molecule, generally with a molecular weight of 500 Daltons or less, which inhibits (e.g., irreversibly) the BTK protein. Exemplary BTK inhibitors include ibrutinib, acalabrutinib (ACP-196), ONO-4059, spebrutinib (CC-292), HM-71224, CG-036806, GDC-0834, ONO-4049, RN-486, SNS-062, TAS-5567, AVL-101, AVL-291, PCI-45261, HCI-1684, PLS-123, and BGB-3l ll. Additional BTK inhibitors for use in the present methods are described, for example, in PCT Publication Nos. WO2014210255, WO2016087994, W02013010380, WO2015061247, WO2013067274, and WO1999054286 and U.S. Patent No. 6,160,010; all incorporated herein by reference in their entirety.

[00207] In some aspects, the PI3K inhibitor is selected from the group of PI3K inhibitors consisting of buparlisib, idelalisib, BYL-719, dactolisib, PF-05212384, pictilisib, copanlisib, copanlisib dihydrochloride, ZSTK-474, GSK-2636771 , duvelisib, GS-9820, PF- 04691502, SAR- 245408, SAR-245409, sonolisib, Archexin, GDC-0032, GDC-0980, apitolisib, pilaralisib, DLBS 1425, PX-866, voxtalisib, AZD-8186, BGT-226, DS-7423, GDC- 0084, GSK-21 26458, INK-l 1 17, SAR-260301 , SF-l 1 26, AMG-319, BAY-1082439, CH- 51 32799, GSK-2269557, P-71 70, PWT-33597, CAL-263, RG-7603, LY-3023414, RP-5264, RV-1729, taselisib, TGR-l 202, GSK-418, INCB-040093, Panulisib, GSK-105961 5, CNX- 1351 , AMG-51 1 , PQR-309, l7beta-Hydroxywortmannin, AEZS-129, AEZS-136, HM- 5016699, IPI-443, ONC-201 , PF-4989216, RP-6503, SF-2626, X-339, XL- 499, PQR-401 , AEZS-132, CZC-24832, KAR-4141 , PQR-31 1 , PQR-316, RP- 5090, VS-5584, X-480, AEZS-126, AS-604850, BAG-956, CAL-130, CZC- 24758, ETP-46321 , ETP-471 87, GNE- 317, GS-548202, HM-032, KAR-l 139, LY-294002, PF-04979064, PI-620, PKI-402, PWT- 143, RP-6530, 3-HOI-BA-Ol , AEZS-134, AS-041 164, AS-252424, AS-605240, AS-605858, AS- 606839, BCCA-621 C, CAY-10505, CH-5033855, CH-51 08134, CUDC-908, CZC-l 9945, D- 106669, D-87503, DPT-NX7, ETP-46444, ETP-46992, GE-21 , GNE-123, GNE-151 , GNE-293, GNE-380, GNE-390, GNE-477, GNE-490, GNE- 493, GNE-614, HMPL-51 8, HS-104, HS-l 06, HS-l 16, HS-173, HS-196, IC- 486068, INK-055, KAR 1 141 , KY-l 2420, Wortmannin, Lin-05, NPT-520-34, PF- 04691503, PF-06465603, PGNX-01 , PGNX-02, PI 620, PI- 103, PI-509, PI-516, PI-540, PIK-75, PWT-458, RO-2492, RP-5152, RP-5237, SB- 20! 5, SB-2312, SB-2343, SHBM-1009, SN 32976, SR-13179, SRX-2523, SRX-2558, SRX- 2626, SRX-3636, SRX-5000, TGR-5237, TGX-221 , UCB-5857, WAY-266175, WAY- 266176, EI-201 , AEZS-131 , AQX-MN100, KCC-TGX, OXY-l 1 1 A, PI-708, PX-2000, and WJD-008.

[00208] It is contemplated that the additional cancer therapy can comprise an antibody, peptide, polypeptide, small molecule inhibitor, siRNA, miRNA or gene therapy which targets, for example, epidermal growth factor receptor (EGFR, EGFR1, ErbB-l, HER1), ErbB-2 (HER2/neu), ErbB-3/HER3, ErbB-4/HER4, EGFR ligand family; insulin-like growth factor receptor (IGFR) family, IGF-binding proteins (IGFBPs), IGFR ligand family (IGF-1R); platelet derived growth factor receptor (PDGFR) family, PDGFR ligand family; fibroblast growth factor receptor (FGFR) family, FGFR ligand family, vascular endothelial growth factor receptor (VEGFR) family, VEGF family; HGF receptor family: TRK receptor family; ephrin (EPH) receptor family; AXL receptor family; leukocyte tyrosine kinase (LTK) receptor family; TIE receptor family, angiopoietin 1, 2; receptor tyrosine kinase-like orphan receptor (ROR) receptor family; discoidin domain receptor (DDR) family; RET receptor family; KLG receptor family; RYK receptor family; MuSK receptor family; Transforming growth factor alpha (TGF- a), TGF-a receptor; Transforming growth factor-beta (TGF-b), TGF-b receptor; Interleukin 13 receptor alpha2 chain (lLl3Ralpha2), Interleukin-6 (IL-6), 1L-6 receptor, Interleukin-4, IL-4 receptor, Cytokine receptors, Class I (hematopoietin family) and Class II (interferon/ 1L- 10 family) receptors, tumor necrosis factor (TNF) family, TNF-a, tumor necrosis factor (TNF) receptor superfamily (TNTRSF), death receptor family, TRAIL-receptor; cancer-testis (CT) antigens, lineage- specific antigens, differentiation antigens, alpha-actinin-4, ARTC1, breakpoint cluster region-Abelson (Bcr-abl) fusion products, B-RAF, caspase-5 (CASP-5), caspase-8 (CASP-8), beta-catenin (CTNNB1), cell division cycle 27 (CDC27), cyclin- dependent kinase 4 (CDK4), CDKN2A, COA-l, dek-can fusion protein, EFTUD-2, Elongation factor 2 (ELF2), Ets variant gene 6/acute myeloid leukemia 1 gene ETS (ETC6-AML1) fusion protein, fibronectin (FN), GPNMB, low density lipid receptor/GDP-L fucose: beta-Dgalactose 2-alpha-Lfucosyltraosferase (LDLR/FUT) fusion protein, HLA-A2, arginine to isoleucine exchange at residue 170 of the alpha-helix of the alpha2-domain in the HLA-A2 gene (HLA- A*20l-Rl70I), MLA-A11, heat shock protein 70-2 mutated (HSP70-2M), KIAA0205, MART2, melanoma ubiquitous mutated 1, 2, 3 (MUM-l, 2, 3), prostatic acid phosphatase (PAP), neo-PAP, Myosin class 1, NFYC, OGT, OS-9, pml-RARalpha fusion protein, PRDX5, PTPRK, K-ras (KRAS2), N-ras (NRAS), HRAS, RBAF600, SIRT2, SNRPD1, SYT-SSX1 or -SSX2 fusion protein, Triosephosphate Isomerase, BAGE, BAGE-l, BAGE-2,3,4,5, GAGE- 1,2, 3, 4, 5, 6, 7, 8, GnT-V (aberrant N-acetyl giucosaminyl transferase V, MGAT5), HERV-K- MEL, KK-LC, KM-HN-l, LAGE, LAGE-l, CTL-recognixed antigen on melanoma (CAMEL), MAGE-A1 (MAGE-l), MAGE-A2, MAGE- A3, MAGE-A4, MAGE-AS, MAGE- A6, MAGE-A8, MAGE-A9, MAGE- A 10, MAGE- Al l, MAGE-A12, MAGE-3, MAGE-B1, MAGE-B2, MAGE-B5, MAGE-B6, MAGE-C1, MAGE-C2, mucin 1 (MUC1), MART- l/Melan-A (MLANA), gplOO, gpl00/Pmell7 (S1LV), tyrosinase (TYR), TRP-l, HAGE, NA- 88, NY-ESO-l, NY-ESO-l/LAGE-2, SAGE, Spl7, SSX-l,2,3,4, TRP2-1NT2, carcino- embryonic antigen (CEA), Kallikfein 4, mammaglobm-A, OA1, prostate specific antigen (PSA), prostate specific membrane antigen, TRP-l/gp75, TRP-2, adipophilin, interferon inducible protein absent in nielanoma 2 (AIM-2), BING-4, CPSF, cyclin Dl, epithelial cell adhesion molecule (Ep-CAM), EpbA3, fibroblast growth factor-5 (FGF-5), glycoprotein 250 (gp250intestinal carboxyl esterase (iCE), alpha-feto protein (AFP), M-CSF, mdm-2, MUCI, p53 (TP53), PBF, FRAME, PSMA, RAGE-l, RNF43, RU2AS, SOX10, STEAP1, survivin (BIRCS), human telomerase reverse transcriptase (hTERT), telomerase, Wilms' tumor gene (WT1), SYCP1, BRDT, SPANX, XAGE, ADAM2, PAGE-5, LIP1, CTAGE-l, CSAGE, MMA1, CAGE, BORIS, HOM-TES-85, AFl5ql4, HCA66I, LDHC, MORC, SGY-l, SPOll, TPX1, NY-SAR-35, FTHLI7, NXF2 TDRD1, TEX 15, FATE, TPTE, immunoglobulin idiotypes, Bence-Jones protein, estrogen receptors (ER), androgen receptors (AR), CD40, CD30, CD20, CD19, CD33, CD4, CD25, CD3, cancer antigen 72-4 (CA 72-4), cancer antigen 15-3 (CA 15-3), cancer antigen 27-29 (CA 27-29), cancer antigen 125 (CA 125), cancer antigen 19-9 (CA 19-9), beta-human chorionic gonadotropin, 1-2 microglobulin, squamous cell carcinoma antigen, neuron-specific enoJase, heat shock protein gp96, GM2, sargramostim, CTLA-4, 707 alanine proline (707-AP), adenocarcinoma antigen recognized by T cells 4 (ART-4), carcinoembryogenic antigen peptide- 1 (CAP-l), calcium-activated chloride channel- 2 (CLCA2), cyclophilin B (Cyp-B), human signet ring tumor-2 (HST-2), Human papilloma vims (HPV) proteins (HPV-E6, HPV-E7, major or minor capsid antigens, others), Epstein-Barr vims (EBV) proteins (EBV latent membrane proteins— LMP1, LMP2; others), Hepatitis B or C vims proteins, and HIV proteins.

VIII. Articles of Manufacture or Kits

[00209] An article of manufacture or a kit is provided comprising a p53 dendritic cell vaccine and preferential CD122/CD132 agonist treatment and, in some embodiments, at least one immune checkpoint inhibitor (e.g., anti-PD-l antibody and/or anti-CTlA-4 antibody) is also provided herein. The article of manufacture or kit can further comprise a package insert comprising instructions for using the p53 dendritic cell vaccine and the CD122/CD132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor to treat or delay progression of cancer in an individual or to enhance immune function of an individual having cancer. Any p53 dendritic cell vaccine and the CD122/CD132 agonist treatment either alone or in combination with at least one immune checkpoint inhibitor described herein may be included in the article of manufacture or kits.

[00210] In some embodiments, the p53 dendritic cell vaccine and the CD 122/CD 132 agonist treatment alone or in combination with at least one immune checkpoint inhibitor (e.g., anti-PD-l antibody and/or anti-CTLA-4 antibody) are in the same container or separate containers. Suitable containers include, for example, bottles, vials, bags and syringes. The container may be formed from a variety of materials such as glass, plastic (such as polyvinyl chloride or polyolefin), or metal alloy (such as stainless steel or hastelloy). In some embodiments, the container holds the formulation and the label on, or associated with, the container may indicate directions for use. The article of manufacture or kit may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use. In some embodiments, the article of manufacture further includes one or more of another agent (e.g., a chemotherapeutic agent, and anti-neoplastic agent). Suitable containers for the one or more agent include, for example, bottles, vials, bags and syringes.

IX. Examples

[00211] The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

Example 1 -Ad-p53 Dendritic Cell (Ad-p53-DC) Vaccine Therapy in Combination with a CD122/CD132 agonist and Immune Checkpoint Inhibition

[00212] The efficacy of an Ad-p53-DC vaccine therapy in combination with a CD 122/CD 132 agonist and an immune checkpoint inhibitor is demonstrated in animal tumor models representative of human cancers.

[00213] Animals tumor inoculation and measurements: Pathogen free six to eight weeks old male C57/B6 mice (Harlan Inc., Indianapolis, IN) are utilized. In the Ad-p53- DC vaccine studies, tumors are generated by injecting animals into the right flank, subcutaneously, with 1 x 10⁶ MC38 tumor cells to form the primary Tumor. Tumor growth is monitored by measuring the length (L) and width (w) of the tumor, and tumor volume calculated using the following formula: volume = (L x w²)/2.

[00214] Viral vectors: Adenovirus 5 vectors with a deleted El region, encoding for expression of wt-p53 (Ad-p53), under the control of the CMV promoter are used (vector construction was described by Zhang et al, 1994). An Eldeleted control adenovirus 5 (Ad-c) is employed to generate control DC cells.

[00215] Ad-p53-DC Vaccine Preparation and Immunization Treatment Procedures: To obtain DCs, bone marrow cells are obtained from the femurs and tibias of C57/B6 mice. Naive BM cells (from both tibias and femurs) are flushed by cutting the ends of the bones first, then applying ice-cold MACS buffer (lx PBS, 2 mM EDTA, 1 % FBS) with a syringe and needle. Once collected, these cells are filtered using a 70 mM cell strainer placed on the top of a 50 mL conical tube. Cells are centrifuged at 1500 rpm for 5 minutes at 4 degrees. Supernatant is removed then pellet is resuspended in 1 mL of ACK buffer. Red blood cells are lysed by leaving the ACK buffer for 5 minutes at RT. After 5 minutes, 20 mL of ice-cold MACS buffer are added, and cells are centrifuged as before. Supernatant is removed, cells are resuspended in 3 mL of ice-cold MACS buffer and counted. [00216] Lineage negative cells are isolated using the Miltenyi mouse lineage cell depletion kit by following manufacturer's guidelines. Once isolated, 20000 cells are put in each well (24-well plate) in a total volume of 1 mL, in complete RPMI with additional mouse Flt- 3L (final concentration: 100 ng / mL) and mouse GM-CSF (final concentration: 10 ng / mL). At day 3, half of the media is removed and replaced with fresh media supplemented with cytokines at the same final concentration than mentioned before. At day 5, all the cells are harvested by taking the supernatant first and then by scraping the wells with a 1 mL syringe plunger after addition of some ice-cold MACS buffer. The cells are counted and seeded in 6- well plates, with 1.5 million cells per well, for a total volume of 3 mL (in complete RPMI and with the same cytokines). Half of the media is changed every 2-3 days and replaced with fresh media containing fresh cytokines. At day 12, the cells are harvested and used for transfection.

[00217] Transfection is performed in 6 well plates: 250 pL (1.5 million) of the cell suspension is added to each well, and on top is added 250 uL of virus suspension (18 000 vp / cell). Both cells and virus are mixed in serum-free RPMI. The plates are then placed for 1 hour in the incubator. After l-hour, complete RPMI containing 20 % FBS and serum- free RPMI are added in order to reach a total volume of 3 mL with 10 % FBS final concentration. Add cytokines as described above. Cells are incubated overnight. The next day, cells are harvested as described above and washed several times with sterile lx PBS. Cells are counted and resuspended at the appropriate concentration in PBS for s.c. vaccination.

[00218] Complete RPMI medium: RPMI 1640 lx from Fisher Scientific (# MT- 10-041-CV) containing L-glutamine and 25 mM HEPES supplemented with 10% FBS, 1% penicillin-streptomycin and 2-mercaptoethanol lx (ThermoFisher Scientific).

[00219] For murine tumor models, murine IL-2 (eBioscience or R&D Systems Minneapolis, MN) is mixed with the S4B6-1 anti-mouse IL2 antibody (Bioxcell, West Lebanon, NH or BD Biosciences) at a molar ratio 2:1 to generate the preferential CD122/CD132 agonist immunocomplex. For studies involving human T cells, human IL-2 is mixed with MAB602 anti-human IL-2 antibody (R&D Systems). The IL-2/S4B6 or IL- 2/MAB602 mAh immunocomplexes are administered intraperitoneally (IP) at 2.5 pg IL2/dose on days 2, 6, and 10. Alternatively, IL-2/S4B6 mAh immunocomplexes were injected on days 2- 6 (1.0 pg IL2/dose). Immunocomplexes are prepared by incubating anti-IL-2 monoclonal with IL-2 for 15 minutes at room temperature. [00220] Immune checkpoint inhibitor therapy. The anti-mouse PD-l antibody (CD279) specifically produced for use in vivo is purchased from BioXcell (catalog # BE0146) as are antibodies to anti-PD-Ll and the immune modulator anti-LAG-3. Anti-PD-l is injected i.p. at a dose of 200 pg/mouse.

[00221] The following treatments are administered and compared: Ad-p53 DC vaccine + preferential CD122/CD132 agonist + immune checkpoint inhibitor; Ad-p53 DC vaccine + preferential CD122/CD132 agonist; Ad-p53 DC vaccine + immune checkpoint inhibitor; preferential CD122/CD132 agonist + immune checkpoint inhibitor; Ad-p53 DC vaccine alone; preferential CD122/CD132 agonist alone, immune checkpoint inhibitor alone. Control groups include treatment with vehicle/saline controls. Treatment efficacy and their synergistic interactions are demonstrated by measurement of tumor volumes and tumor response rates and their statistical analyses by Chi-square analysis, Fisher’s exact test, T test, analysis of variance (ANOVA), Kruskal-Wallis ANOVA and by comparisons of survival using Kaplan-Meier and log rank tests.

[00222] Superior Tumor Responses with Ad-p53 DC vaccine + CD122/132

+ anti-PD-1 treatment. It is generally appreciated that tumor responses with reductions in tumor size following treatment are associated with important therapeutic benefits. FIG. 2A depicts the tumor response rates of reductions in tumor size > 50% for each of the treatment groups. There was an unexpected, surprisingly high rate of tumor responses in the Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group with 87.5 % (7/8 animals) demonstrating a > 50% reduction in tumor volume. None of the other treatment combinations demonstrated synergistic effects as their combined response rates were either similar or lower to each of their monotherapy results. For example, the Ad-p53 DC vaccine and CD122/CD132 treatment combination appeared to have an inhibitory effect with 0% responders. The combination of CD 122/CD 132 and anti-PD-l treatment had no greater response rate than CD 122/CD 132 monotherapy. A Chi-square analysis comparing the treatment groups revealed a highly statistically significant difference between the treatments (p=0.0044). Fisher’s exact test demonstrated a statistically significant increased response rate for tumor size reductions > 50% in the Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group (p-value = 0.0010 by two-sided Fisher’s Exact Test comparing Ad-p53 DC vaccine + CD 122/132 + anti-PD-l treatment group vs. animals in all other Ad-p53 DC vaccine, CD 122/132 and anti-PD-l treatment groups). The specific tumor size reductions in the Ad-p53 DC vaccine + CD 122/132 + anti-PD-l treatment group are shown in FIG. 2B. The overall mean tumor volume reductions for the treatment groups are shown in FIG. 2C. There was a surprisingly high mean tumor volume reduction in the Ad-p53 DC vaccine + CD122/132 + anti-PD-l treatment group (- 67.1%). A statistical analysis of variance (ANOVA) comparison of these mean tumor volume reductions was statistically significant (p-value = 0.0013). Only the Ad-p53 DC vaccine + CD122/132 + anti-PD-l group demonstrated a statistically significant decrease in mean tumor volume reduction vs. the vehicle group (p-value = 0.0005). None of the other treatment combinations demonstrated synergistic or even substantial additive effects as their mean tumor volume reductions were either similar or lower to each of their monotherapy results. The synergistic mean tumor volume reduction for the Ad-p53 DC vaccine + CD 122/132 + anti-PD- 1 treatment group was highly remarkable because the combination of Ad-p53 DC + CD122/132 treatment had a significant inhibitory effect on efficacy.

Example 2 - Ad-p53-DC Vaccine in Combination with CD122/CD132 Agonists for

Treatment of Lung Cancer Patients with Extensive Small Cell Lung Cancer

[00223] The efficacy of an Ad-p53-DC Vaccine in combination with immune augmentation, including CD122/CD132 agonists and/or checkpoint inhibitor antibodies, for patients with extensive lung cancer. The following treatment methods, doses, and schedules are utilized:

[00224] Cells and Method of Collection: The target cells used in this study are autologous dendritic cells (DC), grown in an ex-vivo culture system. The growth conditions have been selected to maximize access of the Ad-p53 viral vector to dendritic cells.

[00225] Leukopheresis Procedures: Patients will not be mobilized before cell collection and will undergo a single leukopheresis procedure performed at least 8 weeks after after the last dose of chemotherapy. The freshly collected leukopheresis cells will undergo a density gradient separation, washed, and the mononuclear cell fraction are harvested and divided into equal fractions for cryopreservation.

[00226] Preparation of Autologous Cells: Approximately one week prior to vaccination, 1 fraction of frozen mononuclear cells (MNC) are thawed, washed and placed in X-VIVO-15 culture medium in tissue culture flasks at a concentration of 1.3-1.7 x 10⁶ cells/cm² available culturing surface, and enriched for monocytes through plastic adherence. After culturing for 2 hours, non-adherent cells are removed and the flask are recharged with fresh culture medium supplemented with 5 ng/mL GM-CSF (Amgen, Thousand Oaks, CA) and 5 ng/mL IL-4 (R&D Systems, Minneapolis MN) for two days, at which time additional supplemented medium are added to the flasks. The cells are incubated for an additional 72 hours. At the completion of incubation, the non-adherent and loosely adherent cells are collected and used for a 2 hour infection with Ad-p53 at a viral particle/cell ratio of 15,000 to 1, which allows for the highest level of p53 expression with the least amount of cytotoxicity (Antonia SJ, et al. Clin Cancer Res 2006). At the end of the incubation, culture medium are added to a concentration of 1 x 10⁶ cells/mL, and cells incubated in flasks for an additional 46 hours, and then harvested, washed and analyzed prior to its used for patient vaccination based on specific release criteria.

[00227] Viral Vectors: Clinical grade Ad-p53 viral vectors encoding for expression of wt-p53 are utilized. A brief description of the vector can be found in Example 1 above.

[00228] Ad-p53 DC Vaccine Administration: The Ad-p53-DC Vaccine is administered three times, at 2-week intervals followed by monthly booster immunizations. The Ad-p53-DC are injected intradermally into 4 separate sites (0.25 mL at each site) in bilateral proximal upper and lower extremities, in the regions of the axillary and inguinal nodal basins.

[00229] CD122/CD132 Agonist Therapy: IL2/anti-IL2 immune complex, or

ILl5/anti-ILl5 immune complex, or an IL15/IL15 Receptor a-IgGl-Fc (ILl5/ILl5Ra-IgGl- Fc) immunocomplex or PEGylated IL2 or PEGylated IL15 or an IL2 mutein or an IL15 mutein is administered in doses ranging between 5-100 ug/kg given either SQ or IV at intervals ranging from weekly to every 2-4 weeks.

[00230] Tmmune Checkpoint Inhibitor Therapy: Patients are treated with nivolumab 1 mg/kg plus ipilimumab 3 mg/kg, every 3 weeks for four cycles, followed by nivolumab 3 mg/kg every 2 weeks.

[00231] Criteria for Efficacy Evaluation:

1. Tumor size is monitored by CT or MRI. The method that provides most accurate measurements is used consistently throughout the study. The measurement is performed on study day 28 or 29 prior to the injections on day 1 of the third cycle if CT or MRI, with scans every 8 weeks. A response must be confirmed by a subsequent determination· RECIST 1.1 criteria is applied. 2. Duration of response is defined as time elapsed from the date of response to time of progression.

3. Progression Free Survival is defined as time elapsed from the day of randomization to the recorded date of progression.

4. Overall survival is defined as time elapsed from day of randomization to death.

5. Efficacy endpoints are correlated with PD-L1, PD-L2, immune cell infiltrates, tumor mutational burden and p53 tumor immunohistochemistry biomarkers in exploratory analyses.

[00232] Safety Evaluation:

1. Adverse events reporting.

2. Physical examinations, vital signs, laboratory tests including CBC, biochemistry, and urinalysis.

[00233] Statistical Methodology: Descriptive statistics are used to summarize all efficacy endpoints. Comparisons are made to previous studies evaluating Ad-p53 DC vaccination alone, immune checkpoint inhibitor therapy alone, Ad-p53 vaccination + immune checkpoint inhibitor therapy, and standard of care treatments.

[00234] All the methods disclosed and claimed herein can be made and executed without undue experimentation considering the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

* * * REFERENCES

The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference.

Aghi et al., Cancer Res., 59:3861-3865, 1999.

Aksentijevich et al. Human Gene Ther. 7:1111, 1996.

Baichwal and Sugden, In: Gene Transfer, Kucherlapati R, ed., New York, Plenum Press, pp.

1 17-148, 1986.

Bailey and Levine, J. Pharm. Biomed. Anal., 11: 285-292, 1993.

Bouvet et al, Cancer Res., 58:2288-2292, 1998.

Buller et al, Cancer Gene Therapy, 9: 553-566, 2002.

Camacho et al. J Clin Oncology, 22(145), 2004.

Carroll et al, Mol Cancer Therapeutics, 1:49-60, 2001.

Carry et al., Bioorganic and Medicinal Chemistry Letters, 23(9):2480-2485, 2013.

Caudell et al., J Immunol., 168:6041-6046, 2002.

Chada et al, Cancer Gene Ther., 13:490-502, 2006.444-448, 1998.

Chada et al, Cancer Gene Ther., 13:490-502, 2006.444-448, 1998.

Chase et al, Nat. Biotechnol., 16:

Chen and Okayama, Mol. Cell. Biol. 7:2745-2752, 1987.

Choi et al. Gene Therapy, 17: 190-201, 2010.

Corrales et al, Cell Reports, 11, 1018-1030, 2015.

Couch et al, Am. Rev. Resp. Dis., 88:394-403, 1963.

Doronin et al, Virology, 305:378-387, 2003.

Fraley et al, Proc. Nat 'l Acad. Sci. 76:3348-3352, 1979.

Fujiwara et al, J Natl Cancer Inst, 86: 1458-1462, 1994.

Ghiringhelli et al, Biomed. J., 38:111-116, 2015.

Graham and Van Der Eb, Virology, 52:456-467, 1973.

Gumani et al, Cancer Chemother Pharmacol., 44(2): 143-151, 1999.

Harland and Weintraub, J. Cell Biol, 101: 1094-1099, 1985.

Hartwell et al, Science, 266: 1821-1828, 1994.

Hurwitz ei a/. Proc Natl Acad Sci. 95(17): 10067-10071, 1998.

Hynes and Ferretti, Methods Enzymol., 235: 606-616, 1994. Iannello et al., J Experimental Medicine, 210(10):2057-2069.

IMLYGIC™ [package insert]. Amgen, Inc., Thousand Oaks, CA; October 2015. Inoue et al, Cancer Letters, 157:105-112, 2000.

International Patent Application No. WO1995001994.

International Patent Application No. WO1998042752.

International Patent Application No. WO1999054286.

International Patent Application No. W02000037504.

International Patent Application No. WO2001014424.

International Patent Application No. W02004058801.

International Patent Publication No. WO 2005/003168.

International Patent Publication No. WO 2005/009465.

International Patent Publication No. WO 2006/003179.

International Patent Publication No. WO 2006/072625.

International Patent Publication No. WO 2006/072626.

International Patent Publication No. WO 2007/042573.

International Patent Publication No. WO 2008/084106.

International Patent Publication No. WO 2010/065939.

International Patent Publication No. WO 2012/071411.

International Patent Publication No. WO 2012/160448.

International Patent Publication No. WO 2012009703.

International Patent Publication No. WO2013067274.

International Patent Publication No. WO1995011986.

International Patent Publication No. W02013010380.

International Patent Publication No. W02014/047350.

International Patent Publication No. WO2014/115080.

International Patent Publication No. WO2014210255.

International Patent Publication No. WO2014/123882.

International Patent Publication No. WO2014/138314.

International Patent Publication No. W02015/016718.

International Patent Publication No. WO2015/027163.

International Patent Publication No. WO2015061247.

International Patent Publication No. W02015/150809.

International Patent Publication No. WO2016/009017.

International Patent Publication No. WO2016087994. Jiang et al, Proc. Natl. Acad. Sci., 93:9160-9165.

Jones et al, J Exp Med. 205(l2):2763-79, 2008.

Kawabe et al, Mol Ther. 6(5):637-44, 2002.

Kawabe et al, Mol. Ther. 6(5): 637-644, 2002.

Kim et al. Journal of the National Cancer Institute, 98(20): 1482-1493, 2006.

Kotin et al, Proc. Natl. Acad. Sci. USA, 87:221 1-2215, 1990.

Kreil, Protein Sci., 4:1666-1669, 1995.

Lichtenstein et al, Int. Rev. Immunol., 23:75-111, 2004.

Liu et al J. Biol. Chem., 270:24864, 1995.

Lui et al., Gene Therapy, 10:292-303, 2003.

Mann et al, Cell, 33:153-159, 1983.

Markowitz et al, J. Virol., 62: 1 120-1 124, 1988.

McLaughlin et al, J. Virol., 62:1963-1973, 1988.

Mellman et al , Nature 480:480- 489, 2011.

Mellman et al , Nature, 480:480- 489, 2011.

Mhashilkar et al, Mol. Medicine 7(4): 271-282, 2001.

Miyahara et al., Cancer Gene Therapy, 13:753-761, 2006.

Mokyr et al, Cancer Res., 58:5301-5304, 1998.

Mole, British Journal of Radiology, 26(305): 234-241, 1953.

MultiVir Inc., Form S-l Registration Statement, U.S. Securities and Exchange Commission, 2015

Muzyczka, Curr. Top. Microbiol Immunol, 158:97-129, 1992.

Nicolas and Rubenstein, In: Vectors : A survey of molecular cloning vectors and their uses, Rodriguez and Denhardt (eds.), Stoneham: Butterworth, pp. 493-513, 1988.

Nicolau and Sene, Biochim. Biophys. Acta, 721:185-190, 1982.

Nicolau el al. Methods Enzymol, 149:157-176, 1987.

Nishikawa et al, Mol. Ther., 9(8): 818-828, 2004b.

Nishikawa et al, Oncogene, 23(42): 7125-7131, 2004a.

Nishizaki M, et al, Clin. Can. Res., 5: 1015-1023, 1999.

Ohashi M, et al., Gut, 44:366-371, 1999.

Okazaki T et al, Intern. Immun. 19(7):813, 2007.

Pardoll, Nat Rev Cancer, 12(4): 252-64, 2012.

Pardoll, Nature Rev Cancer, 12:252-264, 2012.

Patel and Player, Expert Opin Investig Drugs, 17(12): 1885-82, 2008. Philip et al. J. Biol. Chem., 268: 16087, 1993.

Qin, X., et al., Biol Reprod., 56:800-11, l997a.

Qin, X., et al., Biol Reprod., 56:812-20, l997b.

Ridgeway, In: Vectors : A survey of molecular cloning vectors and their uses, Rodriguez RL. Denhardt DT, ed., Stonehar Butterworth, pp. 467-492, 1988.

Ripp t et al, Mol. Cell Biol, 10:689-695, 1990.

Rosenberg et al, Nat Med., 10(19): 909-15, 2004.

Samulski et al, EMBO J. 10:3941-3950, 1991.

Samulski et al, J Virol, 63:3822-3828, 1989.

Sherwood et al., Endocrinology 114:806-13, 1984.

Sobol RE, et al., Chapter 11: Tp53 Gene Therapy for Cancer Treatment and Prevention, NY: Springer Science + Business Media, 2013.

Solodin et al, Biochemistry, 34: 13537, 1995.

Spitz et al., Clin Cancer Research, 2: 1665-1671, 1996.

Swisher et al., Clin Cancer Research, 9:93-101, 2003.

Tatebe S, et al., Int. J Oncol., 15: 229-235, 1999.

Tatebe S, et al., Int. J Oncol., 15: 229-235, 1999.

Tchekmedyian et al, Oncology, 29(l2):990-l002, 2015.

Temin, n: Gene Transfer, Kucherlapati (ed.), New York: Plenum Press, pp. 149-188, 1986. Textor et al, Cancer Res., 7l(l8):5998-6009, 2011.

Thierry et al. Proc. Natl. Acad. Sci., 92(2l):9742-6, 1995.

Timiryasova et al, Biotechniques. 31:534, 6, 8-40, 2001.

Toda et al, Mol. Therapy, 2(4): 324-329, 2000.

Tollefson et al, J. Virol., 70: 2296-2306, 1996.

Top et al, J. Infect. Dis., 124:155-160, 1971.

Tsukamoto et al, Nature Genetics, 9:243, 1995.

U.S. Patent Application No. US20110008369.

U.S. Patent Application No. US2014022021.

U.S. Patent Application No. US20140294898.

U.S. Patent No. 4,797,368.

U.S. Patent No. 4,835,251.

U.S. Patent No. 5,023,321.

U.S. Patent No. 5,139,941.

U.S. Patent No. 5,302,523. U.S. Patent No. 5,384,253.

U.S. Patent No. 5,464,765.

U.S. Patent No. 5,580,859.

U.S. Patent No. 5,589,466.

U.S. Patent No. 5,656,610.

U.S. Patent No. 5,702,932.

U.S. Patent No. 5,736,524.

U.S. Patent No. 5,780,448.

U.S. Patent No. 5,789,215.

U.S. Patent No. 5,811,395.

U.S. Patent No. 5,925,565.

U.S. Patent No. 5,935,819.

U.S. Patent No. 5,945,100.

U.S. Patent No. 5,981,274.

U.S. Patent No. 5,994,136.

U.S. Patent No. 5,994,624.

U.S. Patent No. 6,013,516.

U.S. Patent No. 6,160,010.

U.S. Patent No. 6,207,156.

U.S. Patent No. 7,223,593

U.S. Patent No. 7,537,924

U.S. Patent No. 8,017,114.

U.S. Patent No. 8,119,129.

U.S. Patent No. 8,329,867.

U.S. Patent No. 8,354,509.

U.S. Patent No. 8,846,657.

U.S. Patent No. 9,073,898.

U.S. Patent Publication No. US2011/0039778.

U.S. Patent Publication No. US2015/0202290. Vincent et al, Cancer Res., 70(8):3052-306l, 2010. Waku et al, J Immunol., 165:5884-5890, 2000.

Xu et al. Gene Therapy, 22(3): 31— 40, 2015.

Xu et al., J Gastroenterol., 48(2):203-l3, 2013.

Xue et al, Nature, 445(7l28):656-660, 2007. Young et al., Cancer Gene Ther., 20(9): 531-537, 2013. Zeimet and Marth, The Lancer Oncology, 7:415-422, 2003. Zhang et al, Cancer Gene Ther, 22:17-22, 2015.

Zhang et al, Cancer Gene Ther., 1:5-13, 1994.

Claims

What Is Claimed Is:

1. A method of treating cancer in a subject comprising administering to the subject an effective amount of a p53 dendritic cell vaccine and at least one preferential CD123/CD132 agonist to the subject.

2. The method of claim 1, wherein the p53 dendritic cell vaccine is further defined as a dendritic cell vaccine incorporating p53 as an antigen to generate an anti-p53 immune response.

3. The method of claim 1, wherein the at least one preferential CD123/CD132 agonist preferentially binds to the CD 122/CD 132 receptor complex and has lower affinity binding for the CD25 receptor.

4. The method of claim 3, wherein the at least one preferential CD122/CD132 agonist is an IL-2/anti-IL-2 immune complex, an IL-l5/anti-IL-l5 immune complex, an IL- 15/IL-15 Receptor a-IgGl-Fc (IL-l5/IL-l5Ra-IgGl-Fc) immune complex, PEGylated IL-2, PEGylated IL-15, IL-2 mutein and/or IL-15 mutein.

5. The method of claim 3, wherein the at least one preferential CD122/CD132 agonist is an IL-15 mutant bound to an IL-15 receptor a/IgGl Fc fusion protein.

6. The method of claim 6, wherein the IL-15 mutant is IL-15N72D.

7. The method of claim 5, wherein the IL-15 receptor a/IgGl Fc fusion protein is ALT-

803.

8. The method of claim 3, wherein the IL-15 is pre-complexed with IL-l5Ra to preferentially bind to CD122/CD132.

9. The method of claim 3, wherein the IL-2 receptor agonist is not F42K.

10. The method of claim 1, wherein 2, 3, or 4 CD123/CD132 agonists are administered to the subject.

11. The method of claim 1, wherein the p53 dendritic cell vaccine is prepared using an adenoviral p53 vector.

12. The method of claim 1, wherein the p53 dendritic cell vaccine is administered to non tumor sites.

13. The method of claim 1, wherein the p53 dendritic cell vaccine is administered intradermally, subcutaneously, intramuscularly, intra-peritoneally, orally, by inhalation, or by other forms of mucosal exposure.

14. The method claim 1, wherein the p53 dendritic cell vaccine is administered to tumor sites intratumorally, intraarterially, intravenously, intravascularly, intrapleuraly, intraperitoneally, intratracheally, intrathecally, intramuscularly, endoscopically, intralesionally, percutaneously, subcutaneously, regionally, stereotactically, or by direct injection or perfusion.

15. The method of claim 1, wherein the p53 dendritic cell vaccine is administered via continuous infusion, intratumoral injection, intravenous injection, intra-arterial injection, intra-peritoneal injection, intrapleural injection, or intra-thecal injection.

16. The method of claim 1, wherein the p53 dendritic cell vaccine is administered to non tumor sites and tumor sites.

17. The method of claim 1, wherein the administering to non-tumor sites and tumor sites is sequential.

18. The method of claim 1, wherein the administering to non-tumor sites and tumor sites is concurrent.

19. The method of claim 1, wherein the p53 dendritic cell vaccine is administered after the at least one preferential CD122/CD132 agonist.

20. The method of claim 1, wherein the p53 dendritic cell vaccine is administered before the at least one preferential CD 122/CD 132 agonist.

21. The method of claim 1, wherein the p53 dendritic cell vaccine is administered simultaneously with the at least one preferential CD122/CD132 agonist.

22. The method of claim 1, wherein the p53 dendritic cell vaccine is administered to the subject at non-tumor sites and enhances therapeutic effects or reverses resistance to the preferential CD122/CD132 agonist on distant tumors.

23. The method of claim 1, wherein the p53 dendritic vaccine is administered 2, 3, 4, 5, or 6 times.

24. The method of claim 1, further comprising administering at least one additional p53 vaccine for priming and/or boosting the anti-p53 immune response.

25. The method of claim 24, wherein the at least one additional p53 vaccine is a protein or peptide vaccine, a DNA or RNA vaccine, and/or a genetically engineered vims vaccine expressing p53.

26. The method of claim 24, wherein the at least one additional p53 vaccine for priming and the at least one additional p53 vaccine for boosting comprise the same p53 vaccine constructs.

27. The method of claim 24, wherein the at least one additional p53 vaccine for priming and the at least one additional p53 vaccine for boosting comprise different p53 vaccine constructs.

28. The method of claim 24, wherein the at least one additional p53 vaccine is a single or double stranded DNA vims, RNA virus, adenovims, adeno-associated virus, retrovirus, lentivirus, herpes virus, pox vims, vaccinia virus, vesicular stomatitis virus, polio vims, Newcastle’s Disease vims, myxoma virus, Epstein-Barr virus, influenza vims, reovims, maraba virus, rhabdovims, enadenotucirev or coxsackie vims.

29. The method of claim 1, wherein cancer is melanoma, non-small cell lung, small-cell lung, lung, hepatocarcinoma, retinoblastoma, astrocytoma, glioblastoma, leukemia, neuroblastoma, head, neck, breast, pancreatic, prostate, renal, bone, testicular, ovarian, mesothelioma, cervical, gastrointestinal, urogenital, respiratory tract, hematopoietic, musculoskeletal, neuroendocrine, carcinoma, sarcoma, central nervous system, peripheral nervous system, lymphoma, brain, colon or bladder cancer.

30. The method of claim 1, further comprising administering at least one additional

anticancer treatment.

31. The method of claim 30, wherein the at least one additional anticancer treatment is surgical therapy, chemotherapy, radiation therapy, hormonal therapy, immunotherapy, small molecule therapy, receptor kinase inhibitor therapy, anti- angiogenic therapy, cytokine therapy, cryotherapy, radioablation or a biological therapy.

32. The method of claim 30, wherein the at least one additional anticancer treatment is an immune checkpoint inhibitor.

33. The method of claim 32, wherein the at least one checkpoint inhibitor is selected from an inhibitor of CTLA-4, PD-l, PD-L1, PD-L2, LAG3, BTLA, B7H3, B7H4, TIM3, KIR, or A2aR.

34. The method of claim 33, wherein the at least one checkpoint inhibitor is an anti-PD-l antibody, anti-PD-Ll antibody, anti-PD-L2 antibody, anti-CTLA4 antibody, and/or anti-KIR antibody.

35. The method of claim 34, wherein the anti-PD-l antibody is nivolumab,

pembrolizumab, pidilizumab, AMP-514, REGN2810, CT-011, BMS 936559, MPDL3280A or AMP-224

36. The method of claim 34, wherein the anti-PD-Ll antibody is durvalumab,

atezolizumab, or avelumab.

37. The method of claim 34, wherein the anti-PD-L2 antibody rHIgMl2B7.

38. The method of claim 33, wherein the inhibitor of LAG3 is IMP321 or BMS-986016.

39. The method of claim 33, wherein the inhibitor of A2aR is PBF-509.

40. The method of claim 34, wherein the anti-CTLA-4 antibody is tremelimumab or ipilimumab.

41. The method of claim 34, wherein the anti-KIR antibody is lirilumab.

42. The method of claim 32, wherein more than one checkpoint inhibitor is administered.

43. The method of claim 32, wherein the immune checkpoint inhibitor is administered systemically.

44. The method of claim 30, wherein the at least one additional anticancer treatment is a histone deacetylase (HD AC) inhibitor.

45. The method of claim 44, wherein the HDAC inhibitor is tractinostat.

46. The method of claim 1, further comprising providing an extracellular matrix

degrading protein.

47. The method of claim 46, wherein the extracellular matrix-degrading protein is relaxin, hyaluronidase or decorin.

48. The method of claim 30, wherein the biological therapy is a monoclonal antibody, siRNA, miRNA, antisense oligonucleotide, ribozyme or gene therapy.

49. The method of claim 30, wherein the at least one additional anticancer treatment is an oncolytic virus.

50. The method of claim 49, wherein the oncolytic vims is engineered to express p53, MDA-7, IL-12, at least one heat shock protein, TGF-b inhibitor, and/or IL-10 inhibitor.

51. The method of claim 49, wherein the oncolytic vims is a single- or double-stranded DNA vims, RNA vims, adenovims, adeno-associated vims, retrovims, lentivirus, herpes vims, pox vims, vaccinia virus, vesicular stomatitis vims, polio vims, Newcastle’s Disease virus, Epstein-Barr virus, influenza virus, reoviruses, myxoma vims, maraba vims, rhabdovims, enadenotucirev or coxsackie vims.

52. The method of claim 49, wherein the oncolytic virus is herpes simplex vims.

53. The method of claim 49, wherein the oncolytic virus is engineered to express a

cytokine.

54. The method of claim 53, wherein the cytokine is granulocyte-macrophage colony- stimulating factor (GM-CSF) or IL12.

55. The method of claim 49, wherein the oncolytic virus is further defined as talimogene laherparepvec (T-VEC).

56. The method of claim 49, wherein the at least one oncolytic vims is selected from the group consisting of a single- or double-stranded DNA vims, RNA vims, adenovirus, adeno-associated virus, retrovirus, lentivirus, herpes virus, pox vims, vaccinia vims, vesicular stomatitis virus, polio virus, Newcastle’s Disease virus, Epstein-Barr virus, influenza virus, reovimses, myxoma virus, maraba vims, rhabdovirus,

enadenotucirev, and coxsackie vims.

57. The method of claim 49, wherein the at least one oncolytic vims is Ad5- y CD/mutTKS R39rep -hIL 12 , CAVATAK™, CG0070, DNX-2401, G207, HF10, IMLYGIC™, JX-594, MG1-MA3, MV-NIS, OBP-301, REOLYSIN®, Toca 511, Oncorine (H101), H102, H103, RIGVIR, an adenovirus overexpressing the adenoviral death protein (ADP), T-VEC, a NlL-deleted vaccinia vims, an Elb deleted adenovims, an alpha-fetoprotein (AFP) promoter Ad Ela gene -regulated adenovirus, a modified TERT promoter oncolytic adenovims, an HRE-E2F-TERT hybrid promoter oncolytic adenovims, and/or an adenovims with a Pea3 binding site Ela regulatory sequence deletion and an Elb-l9K clone insertion site.

58. The method of claim 49, wherein the adenovims overexpressing ADP is ViRx007.

59. The method of claim 57, wherein the NlL-deleted vaccinia vims is engineered to express IL-12.

60. The method of claim 30, wherein the at least one additional anticancer treatment is a protein kinase or growth factor signaling pathways inhibitor.

61. The method of claim 60, wherein the protein kinase or growth factor signaling

pathways inhibitor is Afatinib, Axitinib, Bevacizumab, Bosutinib, Cetuximab, Crizotinib, Dasatinib, Erlotinib, Fostamatinib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Mubritinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib,

Ranibizumab, Ruxolitinib, Saracatinib, Sorafenib, Sunitinib, Trastuzumab, Vandetanib, AP23451, Vemurafenib, CAL101, PX-866, LY294002, rapamycin, temsirolimus, everolimus, ridaforolimus, Alvocidib, Genistein, Selumetinib, AZD- 6244, Vatalanib, P1446A-05, AG-024322, ZD1839, P276-00 or GW572016.

62. The method of claim 60, wherein the protein kinase inhibitor is a PI3K inhibitor.

63. The method of claim 62, wherein the PI3K inhibitor is a PI3K delta inhibitor.

64. The method of claim 31, wherein the immunotherapy comprises a cytokine.

65. The method of claim 64, wherein the cytokine is granulocyte macrophage colony- stimulating factor (GM-CSF) or IL12.

66. The method of claim 65, wherein the cytokine is an interleukin and/or an interferon.

67. The method of claim 65, wherein the interleukin is IL-2.

68. The method of claim 65, wherein the interferon is IFNa.

69. The method of claim 31, wherein the immunotherapy comprises a co- stimulatory receptor agonist, a stimulator of innate immune cells, or an activator of innate immunity.

70. The method of claim 69, wherein the co- stimulatory receptor agonist is an anti-OX40 antibody, anti-GITR antibody, anti-CD 137 antibody, anti-CD40 antibody, or an anti- CD27 antibody.

71. The method of claim 69, wherein the stimulator of immune cells is an inhibitor of a cytotoxicity-inhibiting receptor or an agonist of immune stimulating toll like receptors (TLR).

72. The method of claim 69, wherein the cytotoxicity-inhibiting receptor is an inhibitor of NKG2A/CD94 or CD96 TACTILE.

73. The method of claim 71, wherein the TLR agonist is a TLR7 agonist, TLR8 agonist, or TLR9 agonist.

74. The method of claim 31, wherein the immunotherapy comprises a combination of a PD-L1 inhibitor, a 4-1BB agonist, and an 0X40 agonist.

75. The method of claim 31, wherein the immunotherapy comprises a stimulator of

interferon genes (STING) agonist.

76. The method of claim 75, wherein the activator of innate immunity is an IDO inhibitor, TGF inhibitor, or IL-10 inhibitor.

77. The method of claim 31, wherein the chemotherapy comprises a DNA damaging agent.

78. The method of claim 77, wherein the DNA damaging agent is gamma- irradiation, X- rays, UV-irradiation, microwaves, electronic emissions, adriamycin, 5- fluorouracil (5FU), capecitabine, etoposide (VP-16), camptothecin, actinomycin-D, mitomycin C, cisplatin (CDDP), or hydrogen peroxide.

79. A pharmaceutical composition comprising (a) a p53 dendritic cell vaccine; and (b) at least one CD122/CD132 agonist.