Varicella-zoster viruses and methods and materials for using varicella-zoster viruses

Abstract

This document provides methods and materials for treating cancer. For example, this document provides varicella-zoster viruses (VZVs; e.g., recombinant VZVs) having (e.g., designed to have) a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences and methods for using such VZVs as an oncolytic agent (e.g., to treat cancer).

Description

[0001] Attorney Docket No.07039-2319WO1 / 2024-024 VARICELLA-ZOSTER VIRUSES AND METHODS AND MATERIALS FOR USING VARICELLA-ZOSTER VIRUSES CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Patent Application Serial No. 63 / 567,250,filed on March 19, 2024. The disclosure of the prior application is considered part of, and is incorporated by reference in, the disclosure of this application. SEQUENCE LISTING This application contains a Sequence Listing that has been submitted electronically as an XML file named “07039-2319WO1_SL.xml.” The XML file, created on March 7, 2025, is 47,435 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety. TECHNICALFIELDThis document relates to methods and materials for treating cancer. For example, this document provides varicella-zoster viruses (VZVs; e.g., recombinant VZVs) having (e.g., designed to have) a genome that lacks at least a portion of one or more VZV nucleic acid sequences and methods and materials for using such VZVs as an oncolytic agent (e.g., to treat cancer). BACKGROUND Oncolytic viruses are engineered to destroy cancer cells selectively, while simultaneously promoting the immune system to recognize tumor antigens and initiatesystemic antitumor immune responses (Melcher et al., Science, 374(6573):1325-26 (2021)).Among the three human -herpesviruses, herpes simplex virus (HSV) type 1 and type 2 have been extensively studied as anticancer agents, and two HSV-based therapies have received clinical approval: T-VEC, approved in the United States and Europe for the treatment of recurrent melanoma, and DELYTACT, authorized in Japan for glioblastoma therapy (Scanlanet al., Front. Oncol., 12:940019 (2022)). Attorney Docket No.07039-2319WO1 / 2024-024 SUMMARY VZVs, belonging to the group of human -herpesviruses, have not been developed as an oncolytic platform, despite indications from clinical case reports suggesting a potentialassociation between VZV infection and cancer remission (Sorel et al., Front. Microbiol.,9:3170 (2018); Laing et al., J. Infect. Dis., 218(suppl_2):S68-S74 (2018); Khoury et al.,Ther. Adv. Med. Oncol., 14:1-9 (2022); Bierman et al., Cancer, 6(3):591-605 (1953);Kamber et al., Bone Marrow Transplant, 50(4):573-8 (2015); Amirian et al., Cancer Med.,5(6):1352-8 (2016); Al-Anazi et al., Eur. J. Haematol., 75(3):234-40 (2005); and Canniff etal., J. Neurovirol., 17(5):448-54 (2011)). As described herein, VZVs can be used as aplatform for safe and effective oncolytic virotherapy. This document provides methods and materials for treating cancer. For example, this document provides VZVs (e.g., recombinant VZVs) having oncolytic activity. In some cases, one or more VZVs described herein (e.g., one or more VZVs having oncolytic activity) can be used as an oncolytic agent (e.g., to treat cancer). In some cases, one or more VZVs (e.g., recombinant VZVs) described herein can be administered to a mammal having cancer to treat that mammal. As demonstrated herein, VZVs can be used as a safe and effective oncolytic virotherapy to treat cancer (e.g., melanoma). In some cases, one or more VZVs having (e.g., designed to have) a genome that lacks at least a portion of one or more VZV nucleic acid sequences can be used to treat cancer. For example, a VZV can be designed to include a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences (e.g., such that the VZV expresses a reduced or eliminated level of a polypeptide encoded by an endogenous VZV nucleic acid sequence(s)) and administered to a mammal (e.g., a human) having cancer (e.g., melanoma) to stimulate an anti-cancer immune response in that mammal. In some cases, a VZV having (e.g., designed to have) a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences also can include (e.g., can be designed to include) nucleic acid that can encode an adjuvant polypeptide (e.g., a T cell-stimulating factor). For example, a VZV having a genome that (a) lacks at least a portion of one or more endogenous VZV nucleic acid sequences and (b) contains exogenous nucleic acid that can Attorney Docket No.07039-2319WO1 / 2024-024 encode an adjuvant polypeptide can be administered to a mammal (e.g., a human) having cancer (e.g., melanoma) to stimulate an anti-cancer immune response in that mammal. In some cases, one or more VZVs provided herein (e.g., one or more recombinant VZVs) and / or nucleic acid encoding a VZV provided herein (e.g., nucleic acid encoding a recombinant VZV) can be used as anti-cancer agents to reduce the number of cancer cells within a mammal (e.g., a human). In general, one aspect of this document features VZVs having a genome that (a) lacks at least a portion of an endogenous VZV nucleic acid sequence that encodes a deoxyuridine triphosphatase (dUTPase) polypeptide and (b) comprises a nucleic acid sequence encoding a polypeptide heterologous to the VZV. The nucleic acid sequence that encodes the dUTPase polypeptide can be a VZV ORF8. The VZV can be derived from an Ellen strain VZV. The VZV can be derived from an Oka strain VZV. The polypeptide heterologous to the VZV is an adjuvant polypeptide. The adjuvant polypeptide can be an interleukin (IL) 12 p35 polypeptide, an IL12 p40 polypeptide, an IL-15 polypeptide, a CCL4 polypeptide, a CXCL13 polypeptide, or a CCL19 polypeptide. The adjuvant polypeptide can be a single- chain IL12 polypeptide. The polypeptide heterologous to the VZV can be a therapeutic polypeptide. The therapeutic polypeptide can be a pigment epithelium-derived factor (PEDF) polypeptide or a brain-derived neurotropic factor (BDNF) polypeptide. The polypeptide heterologous to the VZV can be a targeting polypeptide. The nucleic acid sequence encoding the polypeptide heterologous to the VZV can be operably linked to a promoter. The promoter can be an inducible promoter. The promoter can be a tet-on promoter, a tet-off promoter, aEF1-HTLV promoter, an RSV promoter, a CMV promoter, or an EF1 promoter.In another aspect, this document features vector including nucleic acid encoding a VZV having a genome that (a) lacks at least a portion of an endogenous VZV nucleic acid sequence that encodes a dUTPase polypeptide and (b) comprises a nucleic acid sequence encoding a polypeptide heterologous to the VZV. The nucleic acid sequence that encodes the dUTPase polypeptide can be a VZV ORF8. The VZV can be derived from an Ellen strain VZV. The VZV can be derived from an Oka strain VZV. The polypeptide heterologous to the VZV is an adjuvant polypeptide. The adjuvant polypeptide can be an IL12 p35 polypeptide, an IL12 p40 polypeptide, an IL-15 polypeptide, a CCL4 polypeptide, a CXCL13 polypeptide, Attorney Docket No.07039-2319WO1 / 2024-024 or a CCL19 polypeptide. The adjuvant polypeptide can be a single-chain IL12 polypeptide. The polypeptide heterologous to the VZV can be a therapeutic polypeptide. The therapeutic polypeptide can be a PEDF polypeptide or a BDNF polypeptide. The polypeptide heterologous to the VZV can be a targeting polypeptide. The nucleic acid sequence encoding the polypeptide heterologous to the VZV can be operably linked to a promoter. The promoter can be an inducible promoter. The promoter can be a tet-on promoter, a tet-off promoter, aEF1-HTLV promoter, an RSV promoter, a CMV promoter, or an EF1 promoter.In another aspect, this document features compositions including a VZV having a genome that (a) lacks at least a portion of an endogenous VZV nucleic acid sequence that encodes a dUTPase polypeptide and (b) comprises a nucleic acid sequence encoding a polypeptide heterologous to the VZV and / or including a vector encoding such a VZV. The nucleic acid sequence that encodes the dUTPase polypeptide can be a VZV ORF8. The VZV can be derived from an Ellen strain VZV. The VZV can be derived from an Oka strain VZV. The polypeptide heterologous to the VZV is an adjuvant polypeptide. The adjuvant polypeptide can be an IL12 p35 polypeptide, an IL12 p40 polypeptide, an IL-15 polypeptide, a CCL4 polypeptide, a CXCL13 polypeptide, or a CCL19 polypeptide. The adjuvant polypeptide can be a single-chain IL12 polypeptide. The polypeptide heterologous to the VZV can be a therapeutic polypeptide. The therapeutic polypeptide can be a PEDF polypeptide or a BDNF polypeptide. The polypeptide heterologous to the VZV can be a targeting polypeptide. The nucleic acid sequence encoding the polypeptide heterologous to the VZV can be operably linked to a promoter. The promoter can be an inducible promoter. The promoter can be a tet-on promoter, a tet-off promoter, a EF1-HTLV promoter, an RSVpromoter, a CMV promoter, or an EF1 promoter.In another aspect, this document features methods for treating a mammal having cancer. The methods can include, or consist essentially of, administering, to a mammal having cancer, a VZV having a genome that (a) lacks at least a portion of an endogenous VZV nucleic acid sequence that encodes a dUTPase polypeptide and (b) comprises a nucleic acid sequence encoding a polypeptide heterologous to the VZV, where the number of cancer cells within the mammal is reduced. The mammal can be a human. The cancer can be a melanoma, a glioma, a breast cancer, a T cell lymphoma, a neuroblastoma, a pancreatic Attorney Docket No.07039-2319WO1 / 2024-024 cancer, a lung cancer, a liver cancer, a kidney cancer, a cervical cancer, a bladder cancer, or a prostate cancer. The administering can include intratumoral injection. The method also can include administering a cancer treatment to the mammal. The cancer treatment can include administering an anti-cancer agent to the mammal. The cancer treatment can include subjecting the mammal to surgery, a radiation therapy, or an adoptive cell transfer therapy. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. BRIEF DESCRIPTION OF THE DRAWINGS Figures 1A-1E. Construction of oncolytic VZV vectors. Figure 1A) Construction of an exemplary VZV-BAC vector. The BAC vector pSG7 encompassing VZV ORF52 and ORF53-54 as homology arms was linearized and transfected into APRE-19 cells on day 1. VZV virus was inoculated on day 3 to enable the site-specific integration of BAC vector into VZV genome within ARPE-19 cells. The plaques of recombinant VZV-BAC virus (identified by GFP expression) were enriched by G418 treatment, and circular VZV-BAC episomes were extracted from infected ARPE-19 cells and transformed into DH10B bacterial cells. The BAC plasmids containing full-length VZV genome were further transferred to the SW102 recombineering bacterial cells for VZV genome reconstruction. Recombinant VZV- BAC constructs were transfected into ARPE-19 or ARPE-19-cre cells for virus reconstitution. Figure 1B) VZV vOka-BAC-bHCG and vOka-BAC-bHCG-gBsyn. The bHCG was inserted between ORF60 and ORF61 of the VZV vaccine Oka strain as a secreted Attorney Docket No.07039-2319WO1 / 2024-024 reporter gene, and the ORF31 (gB) A2453T point mutation was introduced to generate the hyperfusogenic vOka-BAC-bHCG-gBsyn vector. Figure 1C) Plaques of vOka-BAC-bHCG and vOka-BAC-bHCG-gBsyn (MOI = 0.001, 2 dpi). Scale bars = 200 µm. Figure 1D) gB A2453T mutation increased virus syncytium area. vOka-BAC-bHCG and vOka-BAC- bHCG-gBsyn plasmids were transfected into 293T cells (2 µg per 1×106cells), and 3 days post-transfection virus syncytium area was measured and compared (unpaired t-test, n = 5). Figure 1E) Growth curves of recombinant vOka viruses. The APRE-19 cells cultured in six- well plates were infected with viruses on day 0 (1000 PFUs per well), and virus titers were determined on days 1, 3, 5, and 7 (n=3). Virus titers of vOka-BAC-bHCG and vOka-BAC- bHCG-gBsyn on day 3 and day 5 were compared (unpaired t-test). ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 2A-2D. VZV vOka vaccine strain demonstrated potent antitumor activity in a human melanoma xenograft model. Figure 2A) Schematic of the experimental design. Human MeWo melanoma cells were implanted subcutaneously on day 0. Viruses were intratumorally (IT) delivered on days 14, 16, and 18 (5×104PFUs per injection). Figure 2B) Blood bHCG levels. Blood samples were collected on day 5 post-treatment and plasma bHCG levels were compared between treatment groups (n=4, unpaired t-test). Figure 2C) Tumor growth curves (n=10, two-way ANOVA with Tukey's multiple comparisons test). Figure 2D) Animal survival curves (n=10, Log-rank (Mantel-Cox) test). ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 3A-3E. Deletion of ORF8 attenuates VZV replication while not reducingvirus antitumor potency. Figure 3A) Construction of VZV Ellen-BAC- ORF8 and Ellen-BAC- ORF65. Figure 3B) Growth curves of Ellen- ORF8 and Ellen- ORF65. The ARPE-19 cells, dSH-SY5Y neuronal cells, or MeWo cells cultured in six-well plates were infected with viruses on day 0 (1000 PFUs per well), and virus titers were detected on days 3, 6, and 9 (n=3). Virus growth curves were compared between the wild-type backbone virus and mutant viruses (two-way ANOVA with Dunnett's multiple comparisons test (for ARPE-19 cells and MeWo cells) or Tukey's multiple comparisons test (for dSH-SY5Y cells)). Figure 3C) Schematic of the experimental design. Human MeWo melanoma cells were implanted subcutaneously on day 0 (n=8). Viruses were IT delivered on days 20, 22, and 24 (5x104 Attorney Docket No.07039-2319WO1 / 2024-024 PFUs per injection). Figure 3D) Intratumoral virus replication. Tumor slices from different therapy groups (collected on day 5 post-treatment) were stained with anti-VZV ORF68 (gE) antibody (Alexa Flour 594). Representative images are displayed. Scale bars = 100 µm.Figure 3E) Efficacy of VZV Ellen- ORF8 and Ellen- ORF65. Tumor growth curves weredisplayed, and animal survival curves were compared with the Gehan-Breslow-Wilcoxon test. ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 4A-4F. Deletion of ORF8 from VZV improved safety in human MeWo melanoma xenograft model. Figure 4A) Insertion of scIL12 into VZV Ellen genome. TheEF1 / HTLV promoter driven scIL12 was inserted between ORF60 and ORF61. Figure 4B) Invivo testing of Ellen-scIL12, Ellen- ORF8-scIL12, and Ellen- ORF65-scIL12. HumanMeWo melanoma cells were implanted subcutaneously on day 0 (n=9). Viruses were IT delivered on days 20, 22, and 24 (5x104PFUs per injection). Tumor growth curves were displayed. Figure 4C) Blood IL12 concentration. Plasma samples were collected on day 5 post-treatment. IL12 p70 concentrations were determined by ELISA and compared with one- way ANOVA with Tukey's multiple comparisons test. Figure 4D) Deletion of ORF8 led to decreased treatment-related mortality. Treatment or tumor growth related animal deaths were recorded for each group. ND, not detected. Figure 4E) Animal survival curves (Log-rank (Mantel-Cox) test). Figure 4F) Average body weight. ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 5A-5C. VZV with a drug controllable single-chain IL12. Figure 5A) Insertion of tet-off controllable scIL12 into VZV Ellen genome. The ORF8 coding sequence was replaced by tet-off-scIL12 cassette. tTA, tetracycline transactivator protein. The pTight promoter contains a modified tet response element (TRE). Without doxycycline, the tTA protein binds to TRE and activates the expression of scIL12. The introduction of doxycycline leads to its binding with tTA, thus silencing the transactivation of genes controlled by TRE.Figures 5B-5C) Drug controllable expression of IL12 in Ellen- ORF8-tet-off-scIL12. TheMeWo cells were seeded in 24-well plate (2x105cells / well) on day 0 and were culturedovernight. On day 1, cells were infected with Ellen- ORF8-tet-off-scIL12 (200 PFUs perwell), and doxycycline was added to the infected cells at different concentrations (0 µg / mL, 1 µg / mL, or 5 µg / mL). IL12 p70 concentration in the supernatant (Figure 5B) and virus titers Attorney Docket No.07039-2319WO1 / 2024-024 on day 4 (Figure 5C) were detected (n=3). For each virus, IL12 concentrations and virus titers in the wells with or without doxycycline treatment were compared (one-way ANOVA with Tukey's multiple comparisons test). ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 6A-6E. In vivo evaluation of Ellen- ORF8-tet-off-scIL12 in the bilateralB16-F10-nectin1 melanoma syngeneic model. Figure 6A) Schematic of the experimental design. B16-F10-nectin1 cells were implanted subcutaneously on both flanks of C57BL / 6J mice on day 0 (n=10 per group). Viruses were IT delivered (1x105PFUs per injection) into tumors on the right flank on days 11, 13, and 15.. Each mouse received daily intraperitoneal administration of doxycycline (2.5 mg / kg) or vehicle control (PBS) from day 11 to day 17.The KOS- ICP34.5 ICP47-gBsyn-scIL12 virus was HSV-1 KOS strain derived. Figure 6B)Blood IL12 concentration. Plasma samples were collected on day 5 post-treatment (n=5), and IL12 p70 concentrations were compared with one-way ANOVA with Sidak's multiplecomparisons test. Figures 6C-6D) Efficacy of Ellen- ORF8-tet-off-scIL12. Tumor growthcurves were displayed, and animal survival curves were compared with the Log-rank (Mantel-Cox) test. Figure 6E) Average body weight. ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 7A-7G. Oncolytic VZV induces systemic antitumor immune responses in the bilateral B16-F10-nectin1 melanoma syngeneic model. B16-F10-nectin1 cells were implanted subcutaneously on both flanks of C57BL / 6J mice on day 0. Viruses were ITdelivered (1x105 PFUs per injection for Ellen- ORF8 and Ellen- ORF8-tet-off-scIL12) intotumors on the right flank on days 11, 13, and 15. On day 8 post-treatment, spleens, injectedtumors and uninjected tumors were collected. Figures 7A-7B) IFN- Elispot analysis ofvirus-induced antitumor immune responses. A total of 1x105spleen cells per well were co-cultured with 1x104 B16-F10-nectin1 cells or antigen peptide (Trp2180-188 or gp10025-33) for 3days before signal development (n=4). Spot numbers for each group were counted and compared (unpaired t-test). SFUs, spot forming units. Figure 7C) Abundance of CD3- / NK1.1+NK cells in injected and uninjected tumors (n=4, unpaired t-test). Figures 7D-7E) Abundance of CD11b+myeloid cells and CD11b+ / F4 / 80+macrophages in injected and uninjected tumors (n=4, unpaired t-test). Figures 7F-7G) Abundance of CD3+T cells and Attorney Docket No.07039-2319WO1 / 2024-024 CD3+ / Granzyme B+cytotoxic T cells in injected and uninjected tumors (n=4, unpaired t- test). ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 8A-8C. Insertion of NIS or scIL12 into vOka genome prevents virus reconstitution. Figure 8A) Construction of VZV vOka-BAC-NIS and VZV (vOka or Ellen strain based) -BAC-bHCG-scIL12. Figure 8B) Reconstitution of vOka-BAC-bHCG and vOka-BAC-NIS. BAC plasmids were transfected into ARPE-19 cells (2 µg per 6x105cells). Green plaques formed on day 5 post-transfection were counted (n=3). Figure 8C) Reconstitution of vOka-BAC-bHCG-scIL12 and Ellen-BAC-bHCG-scIL12. BAC plasmids were transfected into ARPE-19 or ARPE-19-cre cells (2 µg per 6x105cells). Virus plaques formed on day 5 post-transfection were counted (n=3). scIL12, single chain IL12. Figure 9. Deletion of ORF8 attenuates VZV replication in dSH-SY5Y neuronal cells. The dSH-SY5Y neuronal cells cultured in six-well plates were infected with Ellen-BAC,Ellen-BAC- ORF8, or Ellen-BAC- ORF65 on day 0 (1000 PFUs per well), and virusinfected cells were captured on day 1 and 3. Scale bars = 100 µm. Figures 10A-10C. Potency of single chain IL12. Figure 10A) Structure of mouse single chain IL12 (scIL12). Figures 10B-10C) Potency of IL12 in activating immune cells. The pSelect vector expressing IL12 p40, IL12 p35, or scIL12 was transfected into 293T cells to produce IL12 (group 1: 1 µg pSelect-IL12 p40 plasmid into 1x106293T cells; group 2: 1 µg pSelect-IL12 p35 plasmid into 1x106293T cells; group 3: 1 µg pSelect-IL12 p40 plasmid mixed with 1 µg pSelect-IL12 p35 plasmid into 1x106293T cells; group 4: 1 µg pSelect- scIL12 plasmid into 1x106293T cells). The supernatant samples were collected on day 4 post transfection, and IL12 concentrations (IL12 p40 in group 1, IL12 p35 in group 2, IL12 p70 ingroup 3 and group 4) were determined by ELISA analysis. The Mouse IFN- Elispot analysisinvolved exposing 1x105unstimulated mouse spleen cells per well to various concentrations of IL12 (50 pg / mL, 5 pg / mL, 0.5 pg / mL, or 0.05 pg / mL). After a 3-day co-culture, thenumber of IFN- spots formed in each group was counted and compared (n=4, two-wayANOVA with Dunnett's multiple comparisons test). not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figures 11A-11D. Replication of VZV in B16-F10-nectin1 cells. Figure 11A) VZV entry into B16-F10 cells. The MeWo cells, B16-F10 cells, and B16-F10-nectin1 cells Attorney Docket No.07039-2319WO1 / 2024-024cultured in six-well plates (1x106 cells per well) were infected with Ellen-BAC- ORF8-tet-off-scIL12 virus on day 0 (100 PFUs per well), and agarose overlay was applied to the cell monolayers at 2-hour post infection. On 3 dpi, GFP+cell plaques (from infected MeWo cells) or separated GFP+cells / cell plaques (from infected B16-F10 and B16-F10-nectin1 cells) on 3 dpi was counted and compared (n=3, unpaired t-test). Figures 11B-11C) Virus genome replication and viral gene expression. The MeWo cells and B16-F10-nectin1 cells cultured insix-well plates (1x106 cells per well) were infected with Ellen- ORF8-tet-off-scIL12 viruson day 0 (1000 PFUs per well). On 1 dpi and 3 dpi, infected cells were collected, and total RNA / DNA was extracted. Mock samples without virus infection were collected on 3 dpi as the negative controls. Virus genome copy numbers and mRNA levels of ORF62, ORF28, and ORF68 were determined by qPCR and compared (n=3, multiple t tests). Figure 11D) Synthesis of infectious virions. The MeWo cells and B16-F10-nectin1 cells cultured in six-well plates were infected with Ellen- ORF8-tet-off-scIL12 virus on day 0 (500 PFUs perwell). Infected cells were collected on 3 dpi, and titers of infectious viral particles were determined by plaque formation assay in ARPE-19 cells. not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. DETAILED DESCRIPTION This document provides methods and materials for treating cancer. For example, this document provides VZVs having (e.g., designed to have) a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences. In some cases, a VZV provided herein can have (e.g., can be designed to have) a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such that the VZV expresses a reduced or eliminated level of an endogenous VZV polypeptide encoded by the endogenous VZV nucleic acid sequence(s). For example, a VZV provided herein can have (e.g., can be designed to have) a genome that lacks at least a portion of ORF8 (e.g., such that the VZV expresses a reduced or eliminated level of an endogenous dUTPase polypeptide). For example, a VZV provided herein can have (e.g., can be designed to have) a genome that lacks at least a portion of ORF65 (e.g., such that the VZV expresses a reduced or eliminated level of an endogenous polypeptide that can promote transneuronal transportation of the Attorney Docket No.07039-2319WO1 / 2024-024 virus). Also provided herein are methods for using VZVs provided herein as an oncolytic virotherapy to treat cancer (e.g., melanoma). For example, one or more VZVs provided herein can be administered to a mammal (e.g., a human) having cancer (e.g., melanoma) to stimulate an anti-cancer immune response in that mammal. A VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can have attenuated replication (e.g., can have reduced or eliminated replication competence) as compared to a VZV that is not engineered to lack at least a portion of one or more endogenous VZV nucleic acid sequences. In some cases, a VZV provided here can have a replication cycle that is longer than about 12 hours. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can infect dividing cells. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can infect non- dividing cells. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can be non- pathogenic to (e.g., can have reduced or eliminated virulence in) a mammal being treated as described herein. A VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can be derived from any appropriate VZV. A VZV used to create a VZV provided herein can be any appropriate strain. Examples of VZVs that can be used to make a VZV provided herein include, without limitation, Oka strain VZVs, Ellen strain VZVs, Dumas strain VZVs, VariVax strain VZVs, MSP strain VZVs, BC strain VZVs, and VarilRix strain VZVs. In some cases, a VZV provided herein can be derived from a genome having a nucleic acid sequence set forth in the National Center for Biotechnology Information (NCBI) databases in, for example, Accession No. AB097933, Accession No. JQ972913, Accession No. X04370, Accession No. DQ008355, Accession No. Attorney Docket No.07039-2319WO1 / 2024-024 AY548170, Accession No. AY548171, and Accession No. DQ008354. In some cases, a VZV provided herein can be as described in Example 1. A VZV provided herein can have (e.g., can be designed to have) a genome that lacks at least a portion of any endogenous VZV nucleic acid sequences. In some cases, a VZV provided herein can have (e.g., can be designed to have) a genome that lacks all or a portion of a nucleic acid sequence that is endogenous to the VZV. For example, a VZV provided herein can have (e.g., can be designed to have) a genome that lacks all or a portion of at least of one of the following endogenous VZV nucleic acid sequences: ORF1, ORF2, ORF15, ORF11, ORF12, ORF57, ORF64, ORF8, ORF13, ORF36, ORF59, ORF3, ORF58, and ORF65. In some cases, a VZV provided herein can have (e.g., can be designed to have) a genome that lacks all or a portion of an endogenous ORF8 nucleic acid sequence. When a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) lacks a particular VZV nucleic acid sequence, the VZV can have reduced or eliminated expression of an endogenous polypeptide encoded by that VZV nucleic acid sequence. A VZV provided herein can have reduced or eliminated expression of a polypeptide encoded by any appropriate endogenous VZV nucleic acid sequence. In some cases, a VZV provided herein can have reduced or eliminate expression of an endogenous polypeptide encoded by a VZV non-essential gene. For example, a VZV provided herein can have reduced or eliminate expression of an endogenous VZV membrane polypeptide. For example, a VZV provided herein can have reduced or eliminate expression of an endogenous VZV tegument polypeptide. For example, a VZV provided herein can have reduced or eliminate expression of a polypeptide involved in viral DNA replication (e.g., a dUTPase). For example, a VZV provided herein can have reduced or eliminate expression of a polypeptide involved in transneuronal transportation of the virus. In some cases, a VZV provided herein and having a genome that lacks at least a portion of an endogenous VZV ORF8 can express a reduced or eliminated level of a dUTPase polypeptide. In some cases, a VZV provided herein and having genome that lacks at least a portion of an endogenous VZV ORF65 can express a reduced or eliminated level of a polypeptide that can promote transneuronal transportation of the virus. Attorney Docket No.07039-2319WO1 / 2024-024 Any appropriate method can be used to remove at least a portion of one or more endogenous VZV nucleic acid sequences from the genome of a VZV to produce a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences). In some cases, a deletion of all or a portion of an endogenous VZV nucleic acid sequences can be engineered into a bacterial artificial chromosome (BAC) such that the BAC does not encode a full-length polypeptide encoded by that VZV nucleic acid sequence or a fully functional version of that VZV polypeptide. Such deletions can be any appropriate length that results in the deletion of one or more encoded amino acids and results in a reduction or elimination of the normal function of a polypeptide encoded by that endogenous VZV nucleic acid sequence. For example, portions of a VZV nucleic acid sequence can be removed such that the otherwise encoded polypeptide lacks 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, or more amino acid residues and lacks its normal activity. The portion or portions to be deleted can be removed from any location along the length of the sequence. For example, a portion of an endogenous VZV nucleic acid sequence can be removed at the 5’ end, the 3’ end, or an internal region of a VZV nucleic acid sequence such as a VZV ORF8. In some cases, a VZV provided herein can be produced as described in Example 1. In some cases, a VZV provided herein can have (e.g., can be designed to have) a genome that has one or more additional modifications to least a portion of any endogenous VZV nucleic acid sequences. In some cases, a VZV provided herein can have (e.g., can be designed to have) a genome that encodes a polypeptide (e.g., a recombinant polypeptide) that can enhance cell-to-cell fusion within a mammal (e.g., a human) having cancer (e.g., to promote VZV-mediated immunogenic cell death). A polypeptide that can enhance cell-to- cell fusion within a mammal can be any appropriate polypeptide. In some cases, a polypeptide that can enhance cell-to-cell fusion within a mammal can be a glycoprotein B (gB) polypeptide have one or more (e.g., one, two, three, or more) modifications (e.g., substitutions, additions, and / or deletions) within its amino acid sequence. For example, a polypeptide that can enhance cell-to-cell fusion within a mammal can be a gB polypeptide having a Y881F substitution within its amino acid sequence (a gB (Y881F) polypeptide). Examples of amino acid sequences of an exemplary gB (Y881F) polypeptide as well as an Attorney Docket No.07039-2319WO1 / 2024-024 examples of a nucleic acid that can encode a gB (Y881F) polypeptide can be as shown in Example 2. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can include (e.g., can be designed to include) nucleic acid that can encode one or more polypeptides. For example, a VZV provided herein can include (e.g., can be designed to include) nucleic acid that is heterologous to the VZV such that nucleic acid can encode one or more polypeptides that are heterologous to the VZV. Examples of polypeptides that can be encoded by nucleic acid within a VZV provided herein include, without limitation, therapeutic polypeptides (e.g., immunotherapeutic polypeptides such as immune-stimulating polypeptides), and reporter polypeptides. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can include (e.g., can be designed to include) nucleic acid (e.g., heterologous nucleic acid) that can encode one or more additional polypeptides. In some cases, a polypeptide can be an adjuvant polypeptide. For example, an adjuvant polypeptide can be a T cell-stimulating factor (e.g., polypeptide that can stimulate growth and / or function of a T cell). In another example, an adjuvant polypeptide can stimulate growth and / or function of a natural killer (NK) cell. In some cases, a polypeptide can be a therapeutic polypeptide. In some cases, a polypeptide can be a targeting polypeptide (e.g., a polypeptide fused to a single-chain variable fragment that targets epidermal growth factor receptor (EGFR), which is overexpressed in many cancers, such as in glioma and breast cancer). Examples of polypeptides that can encoded by nucleic acid (e.g., heterologous nucleic acid) present in a VZV provided herein include, without limitation, polypeptides present in an IL12 heterodimer (e.g., p35 (IL12A) polypeptides and p40 (IL12B) polypeptides), IL15 polypeptides, CCL4 polypeptides, CXCL13 polypeptides, CCL19 polypeptides, PEDF polypeptides, and BDNF polypeptides. When a polypeptide that can be encoded by nucleic acid (e.g., heterologous nucleic acid) present in a VZV provided herein includes one or more polypeptides present in an IL12 heterodimer, the polypeptide can be a single-chain IL12 (scIL12) polypeptide (e.g., can be a fusion polypeptide that includes a single p35 polypeptide and single p40 polypeptide). In Attorney Docket No.07039-2319WO1 / 2024-024 some cases, a p35 polypeptide and a p40 polypeptide can be indirectly conjugated to produce a scIL12 polypeptide that can be used in the methods and materials provided herein. For example, a p35 polypeptide and a p40 polypeptide can be connected via a linker such as a peptide linker to produce a scIL12 polypeptide that can be used in the methods and materials provided herein. In some cases, a linker can be a cleavable linker. Examples of linkers that can be used to connect a p35 polypeptide and a p40 polypeptide of a scIL12 polypeptide include, without limitation, elastin linkers, (Gly4Ser)n linkers, and 2A linkers. A polypeptide (e.g., an adjuvant polypeptide such as a T cell-stimulating factor) that can be encoded by nucleic acid (e.g., heterologous nucleic acid) present in a VZV provided herein can include a sequence from any appropriate source. In some cases, a nucleic acid that can be present in a VZV provided herein and that can encode a polypeptide can be from an organism other than VZV. In some cases, a nucleic acid that can be present in a VZV provided herein and that can encode a polypeptide can be a synthetic nucleotide sequence. In some cases, a nucleic acid that can be present in a VZV provided herein and that can encode a polypeptide can be from a mammal (e.g., a human). A polypeptide (e.g., adjuvant polypeptide such as a T cell-stimulating factor) that can be encoded by nucleic acid (e.g., heterologous nucleic acid) present in a VZV provided herein can include any appropriate sequence. Examples of amino acid sequences of polypeptides as well as examples of nucleic acids that can encode polypeptides can be as shown in Example 2. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can include (e.g., can be designed to include) nucleic acid (e.g., heterologous nucleic acid) that can encode one or more reporter polypeptides. For example, a VZV having a genome that (a) lacks at least a portion of one or more endogenous VZV nucleic acid sequences and (b) contains nucleic acid that can encode one or more reporter polypeptides can be administered to a mammal (e.g., a human) having cancer (e.g., melanoma) such that VZV infection can be monitored within that mammal. A VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can include (e.g., can be designed to Attorney Docket No.07039-2319WO1 / 2024-024 include) nucleic acid (e.g., heterologous nucleic acid) that can encode any appropriate reporter polypeptide. Examples of reporter polypeptides include, without limitation, green fluorescent polypeptides (GFPs), human chorionic gonadotropin beta subunit (bHCG) polypeptides, and human sodium iodide symporter (NIS) polypeptides. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) that includes nucleic acid that can encode one or more polypeptides (e.g., a therapeutic polypeptide such as an adjuvant polypeptide and / or a reported polypeptide) also can include one or more regulatory elements operably linked to the nucleic acid that can encode one or more polypeptides. Such regulatory elements can include promoter sequences, enhancer sequences, response elements, signal peptides, internal ribosome entry sequences, polyadenylation signals, terminators, and inducible elements that modulate expression (e.g., transcription or translation) of a nucleic acid. The choice of regulatory element(s) that can be included in a VZV provided herein can depend on several factors, including, without limitation, inducibility, targeting, and the level of expression desired. For example, a promoter can be included in a VZV provided herein to facilitate transcription of nucleic acid encoding an adjuvant polypeptide. For example, a promoter can be included in a VZV provided herein to facilitate transcription of nucleic acid encoding a reporter polypeptide. A promoter can be a naturally occurring promoter or a recombinant promoter (e.g., a chimeric promoter). A promoter can be ubiquitous or inducible (e.g., in the presence of tetracycline (a tet-on promoter) or absence of tetracycline (a tet-off promoter)) and can affect the expression of a nucleic acid encoding a polypeptide in a general manner or in a cell-specific or tissue-specific manner. Examples of promoters that can be used to drive expression of an adjuvant polypeptide (e.g., a T cell stimulating factor such as a scIL12 polypeptide) and / or a reporter polypeptide in cells include, without limitation, tet-on promoters, tet-off promoters (e.g., pTight promoters), EF1-HTLV promoters, RSV promoters,CMV promoters, and EF1 promoters. For example, one or more response elements can beincluded in a VZV provided herein to facilitate inducible expression of nucleic acid (e.g., a nucleic acid encoding an adjuvant polypeptide). A response element can be a naturally occurring response element or a recombinant response element (e.g., a chimeric response element). Examples of response elements that can be used to facilitate inducible expression Attorney Docket No.07039-2319WO1 / 2024-024 of an adjuvant polypeptide (e.g., a T cell stimulating factor such as a scIL12 polypeptide) in cells include, without limitation, tetracycline transactivator (tTA) polypeptides, reverse tetracycline-controlled transactivator (rtTA) polypeptides, and tet response elements (TREs). In cases where a VZV provided herein includes nucleic acid that can encode an adjuvant polypeptide (e.g., a T cell stimulating factor such as a scIL12 polypeptide) and includes nucleic acid that can encode a reporter polypeptide, the nucleic acid that can encode the adjuvant polypeptide and the nucleic acid that can encode the reporter polypeptide can be under the control of the same regulatory elements or can be under the control of separate regulatory elements. For example, a VZV provided herein can include nucleic acid that can encode an adjuvant polypeptide (e.g., a T cell stimulating factor such as a scIL12 polypeptide) that is operably linked to a tet-off promoter and can include nucleic acid that can encode a reporter polypeptide that is operably linked to a EF1-HTLV promoter. As used herein, “operably linked” refers to positioning of a regulatory element in a VZV relative to a nucleic acid encoding a polypeptide in such a way as to permit or facilitate expression of the encoded polypeptide. For example, a VZV can contain a promoter and nucleic acid encoding an adjuvant polypeptide (e.g., a scIL12 polypeptide). In this case, the promoter is operably linked to nucleic acid encoding an adjuvant polypeptide (e.g., a scIL12 polypeptide) such that it drives expression of the adjuvant polypeptide in cells. In some cases, a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can have a genome that (a) lacks at least a portion of one or more endogenous VZV nucleic acid sequences and (b) contains nucleic acid that can encode one or more therapeutic polypeptides. For example, a VZV provided herein can have a genome that (a) lacks at least a portion of an endogenous VZV ORF8 nucleic acid sequence (e.g., such that the VZV expresses a reduced or eliminated level of a dUTPase polypeptide) and (b) contains a nucleic acid sequence including a promoter and nucleic acid that can encode a scIL12 polypeptide such that the promoter can drive expression of the scIL12 polypeptide. In cases where a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) includes (e.g., is designed to include) nucleic acid that can encode one or more therapeutic polypeptides, the Attorney Docket No.07039-2319WO1 / 2024-024 nucleic acid that can encode one or more therapeutic polypeptides can be located in any appropriate location within the genome of the VZV. In some cases, nucleic acid that can encode one or more therapeutic polypeptides present within a genome of a VZV provided herein can be located where the portion of one or more endogenous VZV nucleic acid sequences was removed from. Also provided herein are vectors (e.g., expression vectors) containing nucleic acid encoding a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences). Vectors can carry nucleic acid encoding a VZV provided herein into another cell (e.g., a cancer cell), where it can be replicated and / or expressed. An expression vector, also commonly referred to as an expression construct, is typically a plasmid or vector having an enhancer / promoter region controlling expression of a specific nucleotide sequence. When introduced into a cell, the expression vector can use cellular protein synthesis machinery to produce the virus in the cell. A vector containing nucleic acid encoding a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) can be any appropriate type of expression vector. In some cases, a vector can be a non-viral vector. In some cases, a vector can be a viral vector. When a vector containing nucleic acid encoding a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) is a non-viral vector, any appropriate non-viral vector can be used. In some cases, a non-viral vector can be an expression plasmid (e.g., a cDNA expression vector). Examples non-viral vectors that can be used to deliver nucleic acid encoding a VZV provided herein to a mammal (e.g., a human) include, without limitation, bacterial artificial chromosome (BAC) based vectors and yeast artificial chromosome (YAC) based vectors. When a vector containing nucleic acid encoding a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) is a viral vector, any appropriate viral vector can be used. A viral vector can be derived from a positive-strand virus or a negative-strand virus. A viral vector can be derived from a virus with a DNA genome or a RNA genome. In some cases, a viral vector can infect Attorney Docket No.07039-2319WO1 / 2024-024 dividing cells. In some cases, a viral vector can infect non-dividing cells. Examples virus- based vectors that can be used to deliver nucleic acid encoding a VZV provided herein to a mammal (e.g., a human) include, without limitation, herpes virus amplicon vectors. In some cases, a vector containing nucleic acid encoding a VZV provided herein (e.g., a VZV having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences) also can include one or more regulatory elements operably linked to the nucleic acid encoding the VZV. For example, a promoter can be included in a vector containing nucleic acid encoding a VZV provided herein to facilitate production of the VZV. A promoter can be a naturally occurring promoter or a recombinant promoter (e.g., a chimeric promoter). A promoter can be ubiquitous or inducible and can affect the expression of a nucleic acid encoding a VZV provided herein in a general or tissue-specific manner. Examples of promoters that can be used to drive expression of nucleic acid encoding a VZV provided herein in cells include, without limitation, tet-on promoters, tet-off promoters (e.g., pTight promoters), EF1-HTLV promoters, RSV promoters, CMV promoters, and EF1 promoters. In some cases, one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be formulated into a composition (e.g., a pharmaceutical composition) for administration to a mammal (e.g., a mammal having, or at risk of having, cancer). For example, one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be formulated into a pharmaceutically acceptable composition for administration to a mammal having, or at risk of having, cancer. In some cases, one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be formulated together with one or more pharmaceutically acceptable carriers (additives), excipients, and / or diluents. Examples of pharmaceutically acceptable carriers, excipients, and diluents that can be used in a composition described herein include, without limitation, sucrose, lactose, starch (e.g., starch glycolate), cellulose, cellulose derivatives (e.g., modified celluloses such as microcrystalline cellulose and cellulose ethers like hydroxypropyl cellulose (HPC) and cellulose ether hydroxypropyl methylcellulose (HPMC)), xylitol, sorbitol, mannitol, gelatin, Attorney Docket No.07039-2319WO1 / 2024-024 polymers (e.g., polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), crosslinked polyvinylpyrrolidone (crospovidone), carboxymethyl cellulose, polyethylene- polyoxypropylene-block polymers, and crosslinked sodium carboxymethyl cellulose (croscarmellose sodium)), titanium oxide, azo dyes, silica gel, fumed silica, talc, magnesium carbonate, vegetable stearin, magnesium stearate, aluminum stearate, stearic acid, antioxidants (e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, and selenium), citric acid, sodium citrate, parabens (e.g., methyl paraben and propyl paraben), petrolatum, dimethyl sulfoxide, mineral oil, serum proteins (e.g., human serum albumin), glycine, sorbic acid, potassium sorbate, water, salts or electrolytes (e.g., saline such as phosphate-buffered saline (PBS), protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyacrylates, waxes, wool fat, and lecithin. A composition (e.g., a pharmaceutical composition) containing one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be formulated into any appropriate dosage form. Examples of dosage forms include solid or liquid forms including, without limitation, gums, capsules, tablets (e.g., chewable tablets, and enteric coated tablets), suppositories, liquids, enemas, suspensions, solutions (e.g., sterile solutions), sustained- release formulations, delayed-release formulations, pills, powders, and granules. In some cases, one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be packaged with one or more delivery vehicles. For example, one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be conjugated to delivery vehicle. For example, one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be encapsulated within a delivery vehicle. Delivery vehicles that can be formulated for administering one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein to a mammal (e.g., a human) having cancer include, without limitation, cells (e.g., mesenchymal stem cells such as human Attorney Docket No.07039-2319WO1 / 2024-024 mesenchymal stem cells, epithelial cells such as human epithelial cells, and fibroblasts such as human fibroblasts) and nanoparticles (e.g., lipid nanoparticles). A composition (e.g., a pharmaceutical composition) containing one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be formulated for local or systemic administration. A composition (e.g., a pharmaceutical composition) containing one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be designed for oral or parenteral (including subcutaneous, intratumoral, intramuscular, intravenous, topical, and intradermal) administration. Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions that can contain anti-oxidants, buffers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations can be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets. This document also provides methods and materials for using one or more VZVs provided herein (e.g., one or more VZVs having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences). In some cases, one or more VZVs provided herein can be used for treating a mammal (e.g., a human) having cancer. For example, one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be administered to a mammal having cancer to treat the mammal. In some cases, administering one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein to a mammal (e.g., a human) having cancer can stimulate an anti-cancer immune response in the mammal. Attorney Docket No.07039-2319WO1 / 2024-024 In some cases, the materials and methods provided herein can be used to reduce the size of a cancer in a mammal (e.g., a human). For example, a mammal having cancer (e.g., a human having cancer) in need of treatment thereof can be administered one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein to reduce the size of the cancer in the mammal. In some cases, a mammal (e.g., a human) having cancer can be administered one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein to reduce the number of cancer cells in the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, a mammal (e.g., a human) having cancer can be administered one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein to reduce the volume of one or more tumors in the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, the materials and methods provided herein can be used to improve survival (e.g., recurrence-free survival (RFS) and / or overall survival (OS)) of a mammal (e.g., a human) having cancer. For example, a mammal having cancer (e.g., a human having cancer) in need of treatment thereof can be administered one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein to improve survival of the mammal. For example, a mammal (e.g., a human) having cancer can be administered one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein to improve the survival of a mammal having cancer by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. For example, a mammal (e.g., a human) having cancer can be administered one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein to improve the survival of a mammal having cancer by, for example, at least 6 months (e.g., about 6 months, about 8 months, about 10 months, about 1 year, about 1.5 years, about 2 years, about 2.5 years, about 3 years, about 4 years, about 5 years, or more). Any appropriate mammal having, or at risk of having, cancer can be treated as described herein. Examples of mammals that can have, or can be at risk of having, cancer Attorney Docket No.07039-2319WO1 / 2024-024 and can be treated as described herein (e.g., by administering one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein) include, without limitation, humans, non- human primates such as monkeys, horses, bovine species, porcine species, dogs, cats, mice, and rats. In some cases, a human having cancer can be treated as described herein. A mammal having any type of cancer can be treated as described herein (e.g., by administering one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein). In some cases, a cancer treated as described herein can include one or more solid tumors. In some cases, a cancer treated as described herein can be a hematologic cancer (e.g., a blood cancer). Examples of cancers that can be treated as described herein include, without limitation, melanomas, gliomas, breast cancers, T cell lymphomas, neuroblastomas, pancreatic cancers, lung cancers, liver cancers, kidney cancers, cervical cancers, bladder cancers, and prostate cancers. In some cases, methods described herein also can include identifying a mammal as having cancer. Examples of methods for identifying a mammal as having cancer include, without limitation, physical examination, laboratory tests (e.g., blood and / or urine), biopsy, imaging tests (e.g., X-ray, PET / CT, MRI, and / or ultrasound), nuclear medicine scans (e.g., bone scans), endoscopy, and / or genetic tests. Once identified as having cancer, a mammal can be administered or instructed to self-administer one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein. One or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be administered by any appropriate route (e.g., intratumoral, intraperitoneal, intravenous, intramuscular, subcutaneous, oral, and transdermal) to a mammal. In some cases, one or Attorney Docket No.07039-2319WO1 / 2024-024 more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be administered intratumorally to a mammal (e.g., a human). An effective amount of a composition (e.g., a pharmaceutical composition) containing one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be any amount that can treat the cancer without producing significant toxicity to the mammal. An effective amount of one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be any appropriate amount. In some cases, an effective amount of one or more VZVs provided herein can be from about 1x104plaque forming units (PFUs) to about 1x108PFUs (e.g., from about 1x104PFUs to about 1x107PFUs, from about 1x104PFUs to about 1x106PFUs, from about 1x104PFUs to about 1x105PFUs, from about 1x105PFUs to about 1x108PFUs, from about 1x106PFUs to about 1x108PFUs, from about 1x107PFUs to about 1x108PFUs, from about 1x105PFUs to about 1x107PFUs, from about 1x105PFUs to about 1x106PFUs, or from about 1x106PFUs to about 1x107PFUs). In some cases, an effective amount of nucleic acid encoding one or more VZVs provided herein can be from about 1x108viral genomes per kg of body weight (vg / kg) to about 1x1011vg / kg (e.g., per dose) (e.g., from about 1x108vg / kg to about 1x1010vg / kg, from about 1x108vg / kg to about 1x109vg / kg, from about 1x109vg / kg to about 1x1011vg / kg, from about 1x1010vg / kg to about 1x1011vg / kg, or from about 1x109vg / kg to about 1x1010vg / kg, per dose). The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and severity of the condition (e.g., a cancer) may require an increase or decrease in the actual effective amount administered. The frequency of administration of a composition (e.g., a pharmaceutical composition) containing one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV Attorney Docket No.07039-2319WO1 / 2024-024 provided herein can be any frequency that can treat the cancer without producing significant toxicity to the mammal. For example, the frequency of administration can be from about once a day to about once a week, from about twice a week to about twice a month. The frequency of administration can remain constant or can be variable during the duration of treatment. A course of treatment with a composition containing one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can include rest periods. For example, a composition containing one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be administered daily over a two-week period followed by a two-week rest period, and such a regimen can be repeated multiple times. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, route of administration, and severity of the condition (e.g., a cancer) may require an increase or decrease in administration frequency. An effective duration for administering a composition (e.g., a pharmaceutical composition) containing one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be any duration that treat the cancer without producing significant toxicity to the mammal. For example, the effective duration can vary from several days to several weeks, months, or years. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, route of administration, and severity of the condition being treated. In some cases, one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be used as the sole active agent to treat a mammal (e.g., a human) having cancer. In some cases, one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid Attorney Docket No.07039-2319WO1 / 2024-024 sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein can be administered to a mammal (e.g., a human) having cancer together with one or more (e.g., one, two, three, four, five or more) additional agents and / or therapies used to treat cancer. Examples of additional anti-cancer agents that can be used in combination with one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein include, without limitation, chemotherapeutic agents, targeted therapies, cytotoxic agents, immune checkpoint inhibitors, and any combinations thereof. In cases where one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein are used in combination with additional agents used to treat cancer, the one or more additional agents can be administered at the same time (e.g., in a single composition containing both one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein and containing the one or more additional agents) or independently. For example, one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be administered first, and the one or more additional agents administered second, or vice versa. Examples of therapies that can be used to treat cancer include, without limitation, surgery, radiation therapies, and adoptive cell transfer therapies (e.g., chimeric antigen receptor (CAR) T-cell therapies). In cases where one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein are used in combination with one or more additional therapies used to treat cancer, the one or more additional therapies can be performed at the same time or independently of the administration of one or more VZVs provided herein (e.g., one or more VZVs each having a genome that lacks at least a portion of one or more endogenous VZV nucleic acid sequences such as a VZV ORF8 nucleic acid sequence) and / or nucleic acid encoding a VZV provided herein. For example, the one or more VZVs provided herein and / or nucleic acid encoding a VZV provided herein can be administered before, during, and / or after the one or more additional therapies are performed. In some cases, the size of the cancer (e.g., the number of cancer cells and / or the volume of one or more tumors) present within a mammal and / or the severity of one or more symptoms of the cancer being treated can be monitored. Any appropriate method can be used to determine whether or not the size of the cancer present within a mammal is reduced. For Attorney Docket No.07039-2319WO1 / 2024-024 example, imaging techniques can be used to assess the size of the cancer present within a mammal. The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims. EXAMPLESExample 1: Oncolytic Varicella-Zoster Virus Engineered with ORF8 Deletion and Armed with Drug-Controllable Interleukin-12 This Example describes the design of oncolytic VZV, where the VZV was designed to have a genome that (a) lacks at least a portion of the VZV ORF8 nucleic acid sequence (e.g., such that the VZV expresses a reduced or eliminated level of a dUTPase polypeptide) and (b) contains a nucleic acid that can encode an IL12 polypeptide. For example, oncolytic VZVs were constructed based on the vaccine Oka strain and the laboratory strain Ellen using the bacterial artificial chromosome (BAC) based reverse genetics system. Results Construction of oncolytic VZV vectors The BAC vector based Lambda Red recombineering was used to manipulate the 125kb dsDNA genome of VZV. To achieve this, a BAC vector PSG7 encompassing two homology arms (ORF52 and ORF53-54) was constructed. The homology arms drove the site-specific integration of BAC vector into the VZV genome between ORF52 and ORF53 within ARPE-19 cells (Figure 1A). The recombinant VZV-BAC episome was extracted from infected ARPE-19 cells and then transferred into SW102 bacterial cells, which express the Lambda Red recombineering system (exo, bet and gam). Genetic manipulations of VZV were completed within SW102 cells via homologous recombination, and resulting viral constructs were transfected back into ARPE-19 cells for virus reconstitution. Using this reverse genetics platform, VZV-BAC vectors were constructed based on the vaccine strain vOka and the laboratory strain Ellen. Attorney Docket No.07039-2319WO1 / 2024-024 The vaccine based vOka vector was initially used. The human chorionic gonadotropin beta-subunit (bHCG) gene was inserted between ORF60 and ORF61 as a secreted reporter to monitor viral infection (Figure 1B). A hyperfusogenic mutation of gB (Y881F) was furtherintroduced to the vOka genome for enhancing cell-to-cell fusion (Krabbe et al., Cancers(Basel), 10(7): 216 (2018); and Yang et al., J. Virol., 91(1):e01707-16 (2017)). Increasedsyncytium area formed by vOka-BAC-bHCG-gBsyn (gBY881F) was observed compared to the control virus vOka-BAC-bHCG (Figures 1C-1D) but the growth kinetics of the hyperfusogenic mutant virus was reduced (Figure 1E). VZV demonstrates potent antitumor efficacy in human melanoma xenograft model Following the removal of the LoxP-flanked BAC sequence by cultivating the vOka- bHCG and vOka-bHCG-gBsyn viruses in ARPE-19 cells expressing cre recombinase, their potential for oncolytic activity was evaluated using the MeWo human melanoma xenograft model (Figure 2A). The viruses were administered intratumorally, and the attenuated replication of vOka-bHCG-gBsyn was verified via the monitoring of blood bHCG reporter protein levels (Figure 2B). In comparison to the hyperfusogenic virus, the vOka-bHCG virus exhibited superior anti-tumor efficacy in terms of both inhibiting tumor growth and prolonging the survival of animals (Figures 2C-2D). The gBY881Fmutation was previously demonstrated to interfere with both virus replication and the transcription of host genes(Oliver et al., J. Virol., 91(1):e01613-16 (2017)).Efforts progressed to the incorporation of additional / alternative transgenes into the vOka genome. This encompassed the introduction of the human sodium iodide symporter (NIS) gene as an additional reporter gene, as well as the therapeutic transgene single-chain IL12 (scIL12) (Figure 8A). Upon attempting to reconstitute the recombinant viruses within ARPE-19 cells, it was observed that the incorporation of either NIS or scIL12 transgenes hindered the rescue of vOka constructs. In contrast, the Ellen-BAC-NIS and Ellen-BAC- bHCG-scIL12 viruses were successfully rescued (Figure 8B). Deletion of ORF8 attenuates VZV replication while not reducing virus antitumor potency In pursuit of enhancing the safety profile of viruses derived from the Ellen strain, ORF8 and ORF65, two non-essential genes of VZV, were deleted. ORF8 encodes the viral Attorney Docket No.07039-2319WO1 / 2024-024 deoxyuridine triphosphatase, and ORF65 belongs to the evolutionarily conserved - herpesvirus US9 family. ORF8 and ORF65 were removed from Ellen backbone using the galactokinase (galK)-based positive / negative selection system (Figure 3A). Subsequentassessment of growth curves for the two mutant viruses, Ellen- ORF8 and Ellen- ORF65,indicated a significant reduction of growth kinetics for Ellen- ORF8 in both ARPE epithelialcells and differentiated human neuronal dSH-SY5Y cells but not in MeWo melanoma cells (Figure 3B and Figure 9). On the other hand, the attenuation observed with ORF65 was in dSH-SY5Y neuronal cells but not in ARPE-19 cells or MeWo cells (Figure 3B and Figure 9). The oncolytic characteristics of the two mutant viruses were subsequently assessed using the MeWo melanoma xenograft model (Figure 3C). On day 5 post administration, intratumoral virus replication was observed for both mutant viruses (Figure 3D). Analysis of tumor growth curves and animal survival indicated that neither ORF8 nor ORF65 reduced the antitumor efficacy of VZV in the MeWo model (Figure 3E). Treatment-related mortality occurred in one of the eight mice from the wild type Ellen group (on day 34) but not in other two groups (Figure 3E). The potential for reduced virulence linked to the deletion of ORF8was characterized in vivo. The EF1a-HTLV chimeric promoter driven single chain IL12(scIL12) was inserted between ORF60 and ORF61 in the Ellen-BAC, Ellen-BAC- ORF8,and Ellen-BAC- ORF65 backbones (Figure 4A). After excising the BAC sequence inARPE-19-cre cells, these three viruses were administered to MeWo tumors to assess both their effectiveness and toxicity (Figure 4B). Measurements of IL12 levels in the blood revealed that both ORF8 and ORF65 variants resulted in a decrease in virus replication on day 5 post-treatment, with the ORF8 variant showing a more pronounced attenuation (Figure 4C). Importantly, treatment-related animal deaths were recorded in all the nine mice who received Ellen-scIL12 treatment. However, these numbers decreased to two and six inthe Ellen- ORF8-scIL12 and Ellen- ORF65-scIL12 treatment groups, respectively (Figure4D). Compared to the control and the Ellen-scIL12 groups, the Ellen- ORF8-scIL12 groupexhibited a significant improvement in overall animal survival (Figure 4E-F). Collectively,these findings suggest that Ellen- ORF8 holds promise as an oncolytic VZV vector,possessing attenuated virulence while retaining its antitumor potency in vivo. Attorney Docket No.07039-2319WO1 / 2024-024 Arming VZV with drug controllable single-chain IL12 To combine VZV virotherapy with IL12 immunotherapy and to add a switch for controlling IL12 expression in case of systemic IL12 toxicity, the ORF8 coding sequence in Ellen genome was replaced with the tet-off controllable scIL12, resulting in the generation ofthe Ellen- ORF8-tet-off-scIL12 virus (Figure 5A). The scIL12 encodes IL12 p40 and p35subunits as a fusion protein, and its efficacy has been shown to be comparable to that of the native IL12 p70 complex (Figure 10). Subsequent experimentation revealed that the presence of doxycycline effectively inhibited the expression of scIL12 in infected MeWo cells, while virus replication remained unaffected (Figures 5B-5C). IL12 armed VZV induces systemic antitumor immune responses in the B16-F10-nectin1 syngeneic mouse melanoma model The efficacy of the IL12 armed Ellen virus in an immune competent setting was evaluated. It was found that the B16-F10-nectin1 mouse melanoma cells were permissive tothe Ellen- ORF8-tet-off-scIL12 virus in terms of entry, genome replication, and geneexpression. However, it was impaired in producing infectious virions when compared to the production in human MeWo melanoma cells (Figure 11). In the bilateral B16-F10-nectin1model (Figure 6A), intratumoral administration of the Ellen- ORF8-tet-off-scIL12 virusdemonstrated superior efficacy in inhibiting tumor growth of both injected and uninjectedtumors, thereby extending animal survival in comparison to the Ellen- ORF8 virus (Figures6C-6D). Additionally, intraperitoneal administration of doxycycline significantly reduced the expression of scIL12, subsequently diminishing the anti-tumor effectiveness of Ellen- ORF8-tet-off-scIL12 (Figures 6B-6D). The experiment included the HSV-1 KOS- ICP34.5 ICP47-gBsyn-scIL12 virus, characterized by its hyperfusogenic gB and scIL12expression along with the absence of both ICP34.5 and ICP47, as a comparative control.When given in equal doses (1×105 PFUs per injection), the Ellen- ORF8-tet-off-scIL12virus exhibited better overall antitumor efficacy in comparison to the KOS- ICP34.5 ICP47-gBsyn-scIL12 virus (Figures 6C-6D). After being injected, the oncolyticHSV derived from KOS may cause the lysis of target cells and was then rapidly cleared by Attorney Docket No.07039-2319WO1 / 2024-024 the immune system. No treatment-related toxicity was observed across all treatment groups (Figures 6D-6E). To assess the systemic antitumor immune responses elicited by the Ellen- ORF8-tet-off-scIL12 virus in the bilateral B16-F10-nectin1 model, spleen cells and tumors wereharvested 8 days after treatment. IFN- -Elispot analysis revealed that spleen cells showed apositive immune response against B16-F10-nectin1 tumor cells and tumor antigen peptides(TRP2180-188 / gp10025-33) in the group treated with Ellen- ORF8. The introduction of scIL12in the Ellen- ORF8-tet-off-scIL12 virus further intensified these antitumor immuneresponses (Figures 7A-7B). Additionally, the immune cells that infiltrated both the injected and uninjected tumors were characterized using flow cytometry, applying various immune cell markers. This analysis indicated that the use of the two Ellen viruses did not significantly alter the counts of NK cells within the tumors (Figure 7C). However, there was a notable increase in both tumor-associated CD11b+myeloid cells and CD11b+ / F4 / 80+macrophages ininjected and uninjected tumors post Ellen- ORF8 treatment. The Ellen- ORF8-tet-off-scIL12 virus further significantly increased the levels of CD11b+ / F4 / 80+macrophages in injected tumors and CD11b+myeloid cells in uninjected tumors (Figures 7D-7E). Furthermore, counts of CD3+T cells and CD3+ / granzyme B+cytotoxic T cells were elevatedin both injected and uninjected tumors following Ellen- ORF8 treatment, with the scIL12expression further significantly boosting the abundance of both T cell types (Figures 7F-7G).Overall, these findings suggest that the Ellen- ORF8-tet-off-scIL12 virus successfullytriggered antitumor immune responses, enhancing the presence of CD11b+ / F4 / 80+macrophages, T cells, and granzyme B+cytotoxic T cells within the tumors in the B16-F10- nectin1 model. Together, these results demonstrate that VZVs having a genome that (a) lacks at least a portion of the VZV ORF8 nucleic acid sequence (e.g., such that the VZV expresses a reduced or eliminated level of a dUTPase polypeptide) and (b) contains a nucleic acid that can encode an IL12 polypeptide can be administered to a mammal (e.g., a human) having cancer (e.g., melanoma) to stimulate an anti-cancer immune response in that mammal. Attorney Docket No.07039-2319WO1 / 2024-024 Materials and methods All primers used are as listed in Table 1.

[0002] Attorney Docket No.07039-2319WO1 / 2024-024 Table 1. Primer SequencesSEQ ID123456789101213141516171819 CCTTGTGAT

[0003] Attorney Docket No.07039-2319WO1 / 2024-024 ORF68-R GTGTTCGTGTGCGTTCTCTA

[0004] Attorney Docket No.07039-2319WO1 / 2024-024 Recombinant VZVs To generate a VZV capturing BAC vector, two homology arms were amplified from the vOka episomal DNA using primers No.1-4, and then were cloned into the pSG2 vector using restriction sites BamHI and HindIII, obtained vector was pSG6. The chloramphenicol resistance gene (CmR) cassette in the pSG6 vector was then replaced with the neomycin resistance cassette pPGK-pEM7-NeoR using primers No.5-8. The obtained vector was pSG7. pSG7 was linearized with BamHI and transfected into APRE-19 cells (2 µg of linearized pSG7 was added per 5x105cells), at 24 hours post transfection, live VZV virus (VZV vOka virus from Merk (VARIVAX) or VZV Ellen virus from ATCC (VR-1367)) was added to the transfected cells with multiplicity of infection (MOI)=0.05. At 48 hours post infection, G418 (Geneticin) was added to cells with final concentration of 800 µg / mL to enrich the GFP+infected cells. The circular VZV BAC DNA was extracted from the GFP+cells and transferred to DH10B competent cells (C640003, Invitrogen). BAC colonies containing full length VZV genome were selected. VZV-BAC DNA was then transformed into the SW102 cells (National Cancer Institute Biological Resources Branch). Sequence integrity of the VZV-BACs was verified by next generation sequencing (Genewiz Illumina MiSeq, 2×250 bp configuration). The pCMV-NIS cassette and pCMV-bHCG cassette were amplified from plasmids pcSV40-pCMV-NIS and pcSV40-pCMV-bHCG using primers No. 9 and 10, and then inserted between ORF60 and ORF61 of VZV genome within SW102 cells. The scIL12 expression cassette was amplified and inserted between ORF60 and ORF61 of VZV genome using primers No.11 and 13 or used to replace the AmpR sequence of the pcSV40-pCMV-bHCG cassette inserted using primers No.11 and 12. The mouse scIL12 cassette (mIL12BA) was constructed to encode IL12 p40 and p35 subunits as a fusion protein with an elastin linker between the subunits. The mouse IL12 p40, IL12 p35, and scIL12 coding sequences were then cloned into the pSelect-zeo vector (psetz-mcs, Invivogen) under the control of the EF1 / HTLV composite promoter. The tet-off drug-controllable pTight- scIL12 cassette was amplified from vector pTet-off-mIL12BA-ZeoR, and then was used to replace the ORF8 coding sequence in VZV genome using primers No.14 and 24. The pRSV- tTA cassette was inserted downstream of the pTight-scIL12 cassette using primers No.25 Attorney Docket No.07039-2319WO1 / 2024-024 and 26. Doxycycline (D5207, Sigma) was added to inhibit the expression of the tet-off controllable scIL12 encoded by VZV. The vOka carrying gBY881Fhyperfusogenic mutation was constructed using primers No.15-17. The ORF8 and ORF65 were deleted from VZV genome using primers No.18-20 and Primers No.21-23, respectively. Recombinant VZVs were reconstituted in ARPE-19 cells by FuGENE 6 Transfection Reagent (E2691, Promega), or in ARPE-19-cre cells (for removing the BAC sequence). Cell-free VZV was produced as described elsewhere (Sloutskin et al., J. Virol.Methods,206:128-32 (2014)). In brief, VZV infected ARPE-19 cells in T175 flasks were resuspended in cold PBS-sucrose-glutamate-serum buffer (PSGC) (6 mL PSGC buffer per T175 flask). The cells were then frozen in liquid nitrogen and thawed in 37 °C water bath 2times before being sonicated with a VCX-130 Vibra-Cell sonicator at 1-minute intervals (6 2minutes). The absence of intact cells was verified under microscope. The large cell debriswas then removed by centrifugation at 3000 g for 15 minutes. Virus in the supernatant waspelleted using the Lenti-X Concentrator reagent (631231, Takaro). Virus pellets were resuspended in PBS and were stored at -80°C. For animal studies that require cell preparations as the control group, uninfected ARPE-19 cells underwent processing using the identical procedure as described above for the production of cell-free virus. VZV titers (plaque forming units, PFUs) were determined in ARPE-19 monolayers by plaque formationassay. The HSV-1 KOS- ICP34.5 ICP47-gBsyn-scIL12 virus was grown in Vero cells andpurified as described elsewhere (Jiang et al., Mol. Ther.,32(1):241-56 (2024)).Cell lines The ARPE-19 cells (CRL-2302, ATCC), ARPE-19-cre cells, MeWo cells (HTB-65, ATCC), B16-F10 cells (CRL-6475, ATCC), B16-F10-nectin1 cells, and Vero cells (CCL-81, ATCC) were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% fetal bovine serum (FBS) and 100 units / mL of penicillin / streptomycin. B16-F10-nectin1 cells was constructed to stably express the human nectin1 and support HSV-1 replication. The SH- SY5Y cells (CRL-2266, ATCC) were cultured in DMEM / F12 Medium with 100 units / mL of penicillin / streptomycin and 10% FBS. Differentiated human neuronal dSH-SY5Y cells were obtained by culturing SH-SY5Y cells in DMEM / F12 medium containing 2% FBS and 10 µM Attorney Docket No.07039-2319WO1 / 2024-024 retinoic acid (R2625, Sigma) for 3 days followed by a 5-day differentiation in DMEM / F12 medium supplemented with 100 ng / mL beta-nerve growth factor (NGF, 788506, Biolegend) and 50 ng / mL brain-derived neurotrophic factor (BDNF, 788904, Biolegend). All tumor cell lines were authenticated by IDEXX BioAnalytics. Animal studies For the human MeWo melanoma immune-comprised tumor model, MeWo cells suspended in PBS (5×107cells / mL) were subcutaneously implanted to the right flanks (100 µL per mouse) of 6 weeks old female nude mice (NCRNU-F, Taconic). For the syngeneic mouse B16-F10-nectin1 melanoma tumor model, B16-F10-nectin1 tumor cells suspended in PBS (1×107cells / mL) were subcutaneously implanted to both flanks (100 µL per side of flank) of 5~6 weeks old female C57BL / 6J mice (000664, The Jackson Laboratory). When average tumor diameter reached 5 mm, viruses or control cell preparations were intratumorally (IT) delivered. Totally three doses were given for each mouse at indicated time points. The body weight and tumor volumes were monitored three times per week post virus injection. Blood and tumor samples were collected at indicated time points. Animals were euthanatized when mice have weight loss equal to or exceeding 20% of baseline, tumor burden that equals or exceeds 10% of body weight, development of hind limb paralysis or focal motor deficits. Spleens were collected when animals were euthanatized. For each experimental group, there were no exclusions of animals, experimental units, or data points. ELISA analysis Human hCG-beta ELISA (ELH-hCGb-1, RayBiotech), Mouse IL12 p70 / p40 DuoSet ELISA (DY419-05, R&D systems), and Mouse IL12 p35 ELISA (MBS2515782, MyBioSource) analyses were performed according to the manufacturer's instructions. Immunohistochemistry (IHC) analysis Tumors were fixed in 4% paraformaldehyde (PFA) for 3 days and were dehydrated with 30% sucrose in PBS at 4°C for 2 days. The tumor were then subjected to cryosectioning with a Leica CM1860 Cryostat (tumor slices was set at 40 µm). Tumor slices were penetratedwith 1% Triton X-100 in TBS buffer and stained with 4 ,6-diamidino-2-phenylindole (DAPI, Attorney Docket No.07039-2319WO1 / 2024-024 62248, Invitrogen) and antibodies (Rabbit Anti-Varicella Zoster Virus ORF68 Polyclonal Antibody (VRX-0233J, Creative-Biolabs) and Alexa fluor 594 tagged Goat Anti-Rabbit IgG (ab150080, Abcam)). Images were collected with a Zeiss LSM 780 confocal microscope. Elispot assays IFN- elispot assays were conducted using the Mouse IFN-gamma ELISpot Kit(XEL485, R&D systems) as according to the manufacturer's instructions. Spleens were collected on day 8 post-treatment. Spleen cells suspended in RPMI-1640 Medium (supplemented with 10% FBS, 10 ng / mL mIL2, and 1 µM 2-mercaptoethanol) were loaded to the 96-well plates (1×105spleen cells per well). Anti-tumor cell immunity was analyzed by loading live tumor cells (1×104tumor cells per well) to the spleen cell pre-seeded wells.Anti-tumor antigen immunity was analyzed by loading tumor antigen peptides (Trp2180-188 orgp10025-33, a final concentration of 5µM for each peptide) to the spleen cell pre-seeded wells.After tumor cells, peptides, or IL12s were loaded, the plates were cultured at 5% CO2and 37°C for 3 days before spots were developed and counted. Phytohaemagglutinin P (PHA-P, inh-phap, Invivogen) (100 µg / mL) was added in the positive control wells. Quantitative PCR Total DNA and RNA were extracted from VZV infected cells using the hirt method(Paredes et al., Curr. Protoc. Microbiol., Chapter 14:Unit14E 4 (2012)) and the Rneasy plusmini kit (74134, QIAGEN). The LunaScript RT SuperMix Kit (E3010, NEB was used for first strand cDNA synthesis. Quantitative PCR (qPCR) quantitation of VZV genome copy numbers in total DNA and ORF62, ORF28, and ORF68 mRNA levels in total RNA was performed using the Luna Universal qPCR Master Mix (M3003, NEB) by a Roche LightCycler 96 System. VZV genome was quantified using primers No.27 and 28. ORF62, ORF28, and ORF68 mRNA levels were determined using primers No.29-34. The qPCRthermocycling protocol was set up as: 1. Initial denaturation (95 , 60 seconds, 1 cycle); 2.45 cycles of two-step amplification (95 denaturation for 15 seconds, 60 extension for 30seconds); 3. Melt Curve (60 -95 , 1 cycle). The purified Ellen-BAC plasmid was dilutedas standards to generate the standard curves, and copy numbers were calculated with the LightCycler Software. Attorney Docket No.07039-2319WO1 / 2024-024 Flow Cytometry B16-F10-nectin1 tumors were minced to small pieces and digested for 30 minutes at 37°C with RPMI medium containing 0.1% collagenase type I (SCR103, Sigma), 0.2% dispase type II (D4693, Sigma) and 1% DNAse I (11284932001, Roche). Cell suspension was first passed through a 70 µm cell strainer and then incubated in 3 mL red blood cell lysis buffer (420301, Biolegend) per tumor sample. Cell suspension was pelleted by centrifugation for 5 minutes at 350× g and resuspended in FACS buffer (1 mM EDTA, and 2% FBS in Ca / Mg2+free Hanks Buffer) before flow cytometry analysis. To determine the abundance of tumor-associated myeloid cells, macrophages, NK cells, and T cells in B16-F10-nectin1 tumors, dissociated tumor cells were stained with Zombie Aqua dye (423101, Biolegend) and fixed with 4% paraformaldehyde (PFA). For analyzing myeloid cells and macrophages, cells were stained with Pacific Blue tagged anti- mouse / human CD11b antibody (101223, Biolegend) and Alexa Fluor 647 tagged anti-mouse F4 / 80 antibody (123122, Biolegend), or with isotype controls. For phenotyping NK cells and T cells, cells were stained with Pacific Blue anti-mouse NK-1.1 Antibody (108722, Biolegend), Alexa Fluor 647 tagged anti-mouse CD3 antibody (100209, Biolegend), and FITC tagged anti-mouse Granzyme B Antibody (372206, Biolegend), or with isotype controls. Stained cells were analyzed with a BD LSRFortessa™ X-20 Cell Analyzer. Statistics Statistical analyses of data were performed using GraphPad Prism 8 software. Values were presented as mean ± Standard Error of Mean (SEM). All data sets passed Normality test (Shapiro-Wilk test or Kolmogorov-Smirnov test). Data sets were subjected to one-way ANOVA with Tukey's multiple comparisons test or Sidak's multiple comparisons test, two- way ANOVA (Tukey's multiple comparisons test or Dunnett's multiple comparisons test), unpaired t-test (two tailed), or multiple t tests to calculate p-values for group comparisons.p < 0.05 was considered significant. ns, not significant, *p<0.05, **p<0.01, ***p<0.001,****p<0.0001. Attorney Docket No.07039-2319WO1 / 2024-024 Example 2: Exemplary Sequences This Example provides sequences of exemplary polypeptides that can be present in a VZV provided herein and nucleic acid sequences that can encoded such polypeptides. Human IL12 p40 polypeptide sequence MCHQQLVISWFSLVFLASPLVAIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTL DQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPK NKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKE YEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKN SRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASISVRA QDRYYSSSWSEWASVPCS (SEQ ID NO:35) nucleic acid sequence encoding a human p40 polypeptide ATGTGTCACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTCCCCTCGT GGCCATATGGGAACTGAAGAAAGATGTTTATGTCGTAGAATTGGATTGGTATCCGGATGCCC CTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTGAAGAAGATGGTATCACCTGGACCTTG GACCAGAGCAGTGAGGTCTTAGGCTCTGGCAAAACCCTGACCATCCAAGTCAAAGAGTTTGG AGATGCTGGCCAGTACACCTGTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGC TTCACAAAAAGGAAGATGGAATTTGGTCCACTGATATTTTAAAGGACCAGAAAGAACCCAAA AATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCACCTGCTGGTGGCT GACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGCAGCAGAGGCTCTTCTGACCCCC AAGGGGTGACGTGCGGAGCTGCTACACTCTCTGCAGAGAGAGTCAGAGGGGACAACAAGGAG TATGAGTACTCAGTGGAGTGCCAGGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCC CATTGAGGTCATGGTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTCT TCATCAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGCCATTAAAGAAT TCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTGGAGTACTCCACATTCCTACTT CTCCCTGACATTCTGCGTTCAGGTCCAGGGCAAGAGCAAGAGAGAAAAGAAAGATAGAGTCT TCACGGACAAGACCTCAGCCACGGTCATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCC CAGGACCGCTACTATAGCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGTTAA (SEQ ID NO:36) Attorney Docket No.07039-2319WO1 / 2024-024 Human p35 polypeptide sequence MWPPGSASQPPPSPAAATGLHPAARPVSLQCRLSMCPARSLLLVATLVLLDHLSLARNLPVA TPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLEL TKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQ IFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMS YLNAS (SEQ ID NO:37) nucleic acid sequence encoding a human p35 polypeptide ATGTGGCCCCCTGGGTCAGCCTCCCAGCCACCGCCCTCACCTGCCGCGGCCACAGGTCTGCA TCCAGCGGCTCGCCCTGTGTCCCTGCAGTGCCGGCTCAGCATGTGTCCAGCGCGCAGCCTCC TCCTTGTGGCTACCCTGGTCCTCCTGGACCACCTCAGTTTGGCCAGAAACCTCCCCGTGGCC ACTCCAGACCCAGGAATGTTCCCATGCCTTCACCACTCCCAAAACCTGCTGAGGGCCGTCAG CAACATGCTCCAGAAGGCCAGACAAACTCTAGAATTTTACCCTTGCACTTCTGAAGAGATTG ATCATGAAGATATCACAAAAGATAAAACCAGCACAGTGGAGGCCTGTTTACCATTGGAATTA ACCAAGAATGAGAGTTGCCTAAATTCCAGAGAGACCTCTTTCATAACTAATGGGAGTTGCCT GGCCTCCAGAAAGACCTCTTTTATGATGGCCCTGTGCCTTAGTAGTATTTATGAAGACTTGA AGATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTTCTGATGGATCCTAAGAGGCAG ATCTTTCTAGATCAAAACATGCTGGCAGTTATTGATGAGCTGATGCAGGCCCTGAATTTCAA CAGTGAGACTGTGCCACAAAAATCCTCCCTTGAAGAACCGGATTTTTATAAAACTAAAATCA AGCTCTGCATACTTCTTCATGCTTTCAGAATTCGGGCAGTGACTATTGATAGAGTGATGAGC TATCTGAATGCTTCCTAA (SEQ ID NO:38) Human scIL12 polypeptide sequence MCHQQLVISWFSLVFLASPLVAIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTL DQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPK Attorney Docket No.07039-2319WO1 / 2024-024 NKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKE YEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKN SRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASISVRA QDRYYSSSWSEWASVPCSVPGVGVPGVGARNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKA RQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTS FMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQ KSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMSYLNAS (SEQ ID NO:39) nucleic acid sequence encoding a human scIL12 polypeptide ATGTGTCACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTCCCCTCGT GGCCATATGGGAACTGAAGAAAGATGTTTATGTCGTAGAATTGGATTGGTATCCGGATGCCC CTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTGAAGAAGATGGTATCACCTGGACCTTG GACCAGAGCAGTGAGGTCTTAGGCTCTGGCAAAACCCTGACCATCCAAGTCAAAGAGTTTGG AGATGCTGGCCAGTACACCTGTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGC TTCACAAAAAGGAAGATGGAATTTGGTCCACTGATATTTTAAAGGACCAGAAAGAACCCAAA AATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCACCTGCTGGTGGCT GACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGCAGCAGAGGCTCTTCTGACCCCC AAGGGGTGACGTGCGGAGCTGCTACACTCTCTGCAGAGAGAGTCAGAGGGGACAACAAGGAG TATGAGTACTCAGTGGAGTGCCAGGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCC CATTGAGGTCATGGTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTCT TCATCAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGCCATTAAAGAAT TCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTGGAGTACTCCACATTCCTACTT CTCCCTGACATTCTGCGTTCAGGTCCAGGGCAAGAGCAAGAGAGAAAAGAAAGATAGAGTCT TCACGGACAAGACCTCAGCCACGGTCATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCC CAGGACCGCTACTATAGCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGTGTTCCTGG AGTAGGGGTACCTGGGGTGGGCGCCAGAAACCTCCCCGTGGCCACTCCAGACCCAGGAATGT TCCCATGCCTTCACCACTCCCAAAACCTGCTGAGGGCCGTCAGCAACATGCTCCAGAAGGCC AGACAAACTCTAGAATTTTACCCTTGCACTTCTGAAGAGATTGATCATGAAGATATCACAAA AGATAAAACCAGCACAGTGGAGGCCTGTTTACCATTGGAATTAACCAAGAATGAGAGTTGCC Attorney Docket No.07039-2319WO1 / 2024-024 TAAATTCCAGAGAGACCTCTTTCATAACTAATGGGAGTTGCCTGGCCTCCAGAAAGACCTCT TTTATGATGGCCCTGTGCCTTAGTAGTATTTATGAAGACTTGAAGATGTACCAGGTGGAGTT CAAGACCATGAATGCAAAGCTGCTGATGGATCCTAAGAGGCAGATCTTTCTAGATCAAAACA TGCTGGCAGTTATTGATGAGCTGATGCAGGCCCTGAATTTCAACAGTGAGACTGTGCCACAA AAATCCTCCCTTGAAGAACCGGATTTTTATAAAACTAAAATCAAGCTCTGCATACTTCTTCA TGCTTTCAGAATTCGGGCAGTGACTATTGATAGAGTGATGAGCTATCTGAATGCTTCCTAA (SEQ ID NO:40) gB (Y881F) polypeptide sequence MSPCGYYSKWRNRDRPEYRRNLRFRRFFSSIHPNAAAGSGFNGPGVFITSVTGVWLCFLCIF SMFVTAVVSVSPSSFYESLQVEPTQSEDITRSAHLGDGDEIREAIHKSQDAETKPTFYVCPP PTGSTIVRLEPTRTCPDYHLGKNFTEGIAVVYKENIAAYKFKATVYYKDVIVSTAWAGSSYT QITNRYADRVPIPVSEITDTIDKFGKCSSKATYVRNNHKVEAFNEDKNPQDMPLIASKYNSV GSKAWHTTNDTYMVAGTPGTYRTGTSVNCIIEEVEARSIFPYDSFGLSTGDIIYMSPFFGLR DGAYREHSNYAMDRFHQFEGYRQRDLDTRALLEPAARNFLVTPHLTVGWNWKPKRTEVCSLV KWREVEDVVRDEYAHNFRFTMKTLSTTFISETNEFNLNQIHLSQCVKEEARAIINRIYTTRY NSSHVRTGDIQTYLARGGFVVVFQPLLSNSLARLYLQELVRENTNHSPQKHPTRNTRSRRSV PVELRANRTITTTSSVEFAMLQFTYDHIQEHVNEMLARISSSWCQLQNRERALWSGLFPINP SALASTILDQRVKARILGDVISVSNCPELGSDTRIILQNSMRVSGSTTRCYSRPLISIVSLN GSGTVEGQLGTDNELIMSRDLLEPCVANHKRYFLFGHHYVYYEDYRYVREIAVHDVGMISTY VDLNLTLLKDREFMPLQVYTRDELRDTGLLDYSEIQRRNQMHSLRFYDIDKVVQYDSGTAIM QGMAQFFQGLGTAGQAVGHVVLGATGALLSTVHGFTTFLSNPFGALAVGLLVLAGLVAAFFA YRYVLKLKTSPMKALYPLTTKGLKQLPEGMDPFAEKPNATDTPIEEIGDSQNTEPSVNSGFD PDKFREAQEMIKFMTLVSAAERQESKARKKNKTSALLTSRLTGLALRNRRGYSRVRTENVTG V (SEQ ID NO:41) Attorney Docket No.07039-2319WO1 / 2024-024 nucleic acid sequence encoding a gB (Y881F) polypeptide ATGTCCCCTTGTGGCTATTATTCAAAGTGGAGAAACAGGGATCGACCAGAATACCGTCGTAA TCTACGATTCAGACGTTTTTTCTCTTCTATACACCCTAATGCAGCGGCTGGCTCCGGATTCA ACGGACCCGGCGTTTTCATAACCTCCGTTACGGGGGTGTGGTTATGCTTTTTATGCATATTT TCTATGTTTGTTACGGCGGTTGTGTCGGTCTCTCCAAGCTCGTTTTATGAGAGTTTACAAGT AGAGCCCACACAATCAGAAGATATAACCCGGTCTGCTCATCTGGGCGATGGTGATGAAATCA GAGAAGCTATACACAAGTCCCAGGACGCCGAAACAAAACCCACGTTTTACGTCTGCCCACCG CCAACAGGCTCCACAATCGTACGATTAGAACCAACTCGGACATGTCCGGATTATCACCTTGG TAAAAACTTTACAGAGGGTATTGCTGTTGTTTATAAAGAAAACATTGCAGCGTACAAGTTTA AGGCGACGGTATATTACAAAGATGTTATCGTTAGCACGGCGTGGGCCGGAAGTTCTTATACG CAAATTACTAATAGATATGCGGATAGGGTACCAATTCCCGTTTCAGAGATCACGGACACCAT TGATAAGTTTGGCAAGTGTTCTTCTAAAGCAACGTACGTACGAAATAACCACAAAGTTGAAG CCTTTAATGAGGATAAAAATCCACAGGATATGCCTCTAATCGCATCAAAATATAATTCTGTG GGATCCAAAGCATGGCATACTACCAATGACACGTACATGGTTGCCGGAACCCCCGGAACATA TAGGACGGGCACGTCGGTGAATTGCATCATTGAGGAAGTTGAAGCCAGATCAATATTCCCTT ATGATAGTTTTGGACTTTCCACGGGAGATATAATATACATGTCCCCGTTTTTTGGCCTACGG GATGGTGCATACAGAGAACATTCCAATTATGCAATGGATCGTTTTCACCAGTTTGAGGGTTA TAGACAAAGGGATCTTGACACTAGAGCATTACTGGAACCTGCAGCGCGGAACTTTTTAGTCA CGCCTCATTTAACGGTTGGTTGGAACTGGAAGCCAAAACGAACGGAAGTTTGTTCGCTTGTC AAGTGGCGTGAGGTTGAAGACGTAGTTCGCGATGAGTATGCACACAATTTTCGCTTTACAAT GAAAACACTTTCTACCACGTTTATAAGTGAAACAAACGAGTTTAATCTTAACCAAATCCATC TCAGTCAATGTGTAAAGGAGGAAGCCCGGGCTATTATTAACCGGATCTATACAACCAGATAC AACTCATCTCATGTTAGAACCGGGGATATCCAGACCTACCTTGCCAGAGGGGGGTTTGTTGT GGTGTTTCAACCCCTGCTGAGCAATTCCCTCGCCCGTCTCTATCTCCAAGAATTGGTCCGTG AAAACACTAATCATTCACCACAAAAACACCCGACTCGAAATACCAGATCCCGACGAAGCGTG CCAGTTGAGTTGCGTGCCAATAGAACAATAACAACCACCTCATCGGTGGAATTTGCTATGCT CCAGTTTACATATGACCACATTCAAGAGCATGTTAATGAAATGTTGGCACGTATCTCCTCGT CGTGGTGCCAGCTACAAAATCGCGAACGCGCCCTTTGGAGCGGACTATTTCCAATTAACCCA AGTGCTTTAGCGAGCACCATTTTGGATCAACGTGTTAAAGCTCGTATTCTCGGCGACGTTAT CTCCGTTTCTAATTGTCCAGAACTGGGATCAGATACACGCATTATACTTCAAAACTCTATGA Attorney Docket No.07039-2319WO1 / 2024-024 GGGTATCTGGTAGTACTACGCGTTGTTATAGCCGTCCTTTAATTTCAATAGTTAGTTTAAAT GGGTCCGGGACGGTGGAGGGCCAGCTTGGAACAGATAACGAGTTAATTATGTCCAGAGATCT GTTAGAACCATGCGTGGCTAATCACAAGCGATATTTTCTATTTGGGCATCACTACGTATATT ATGAGGATTATCGTTACGTCCGTGAAATCGCAGTCCATGATGTGGGAATGATTAGCACTTAC GTAGATTTAAACTTAACACTTCTTAAAGATAGAGAGTTTATGCCGCTGCAAGTATATACAAG AGACGAGCTGCGGGATACAGGATTACTAGACTACAGTGAAATTCAACGCCGAAATCAAATGC ATTCGCTGCGTTTTTATGACATAGACAAGGTTGTGCAATATGATAGCGGAACGGCCATTATG CAGGGCATGGCTCAGTTTTTCCAGGGACTTGGGACCGCGGGCCAGGCCGTTGGACATGTGGT TCTTGGGGCCACGGGAGCGCTGCTTTCCACCGTACACGGATTTACCACGTTTTTATCTAACC CATTTGGGGCATTGGCCGTGGGATTATTGGTTTTGGCGGGACTGGTAGCGGCCTTTTTTGCG TACCGGTACGTGCTTAAACTTAAAACAAGCCCGATGAAGGCATTATATCCACTCACAACCAA GGGGTTAAAACAGTTACCGGAAGGAATGGATCCCTTTGCCGAGAAACCCAACGCTACTGATA CCCCAATAGAAGAAATTGGCGACTCACAAAACACTGAACCGTCGGTAAATAGCGGGTTTGAT CCCGATAAATTTCGAGAAGCCCAGGAAATGATTAAATtTATGACGTTAGTATCTGCGGCTGA GCGCCAAGAATCTAAAGCCCGCAAAAAAAATAAGACTAGCGCCCTTTTAACTTCACGTCTTA CCGGCCTTGCTTTACGAAATCGCCGAGGATACTCCCGTGTTCGCACCGAGAATGTAACGGGG GTGTAA (SEQ ID NO:42) OTHEREMBODIMENTSIt is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

Attorney Docket No.07039-2319WO1 / 2024-024 WHAT IS CLAIMED IS:

1. A varicella-zoster virus (VZV), wherein said VZV comprises a genome that (a) lacksat least a portion of an endogenous VZV nucleic acid sequence that encodes a deoxyuridine triphosphatase (dUTPase) polypeptide and (b) comprises a nucleic acid sequence encoding a polypeptide heterologous to said VZV.

2. The VZV of claim 1, wherein said nucleic acid sequence that encodes said dUTPasepolypeptide is a VZV ORF8.

3. The VZV of any one of claims 1-2, wherein said VZV is derived from an Ellen strainVZV.

4. The VZV of any one of claims 1-2, wherein said VZV is derived from an Oka strainVZV.

5. The VZV of any one of claims 1-4, wherein said polypeptide heterologous to saidVZV is an adjuvant polypeptide.

6. The VZV of claim 5, wherein said adjuvant polypeptide is selected from the groupconsisting of an interleukin (IL) 12 p35 polypeptide, an IL12 p40 polypeptide, an IL-15 polypeptide, a CCL4 polypeptide, a CXCL13 polypeptide, and a CCL19 polypeptide.

7. The VZV of claim 5, wherein said adjuvant polypeptide is a single-chain IL12polypeptide.

8. The VZV of any one of claims 1-4, wherein said polypeptide heterologous to saidVZV is a therapeutic polypeptide.Attorney Docket No.07039-2319WO1 / 2024-0249. The VZV of claim 8, wherein said therapeutic polypeptide is selected from the groupconsisting of a pigment epithelium-derived factor (PEDF) polypeptide and a brain-derived neurotropic factor (BDNF) polypeptide.

10. The VZV of any one of claims 1-4, wherein said polypeptide heterologous to said VZV is a targeting polypeptide.

11. The VZV of any one of claims 1-10, wherein said nucleic acid sequence encoding said polypeptide heterologous to said VZV is operably linked to a promoter.

12. The VZV of claim 11, wherein said promoter is an inducible promoter.

13. The VZV of claim 11, wherein said promoter is selected from the group consisting of a tet-on promoter, a tet-off promoter, a EF1-HTLV promoter, an RSV promoter, a CMVpromoter, and an EF1 promoter.

14. A vector comprising nucleic acid encoding a VZV of any one of claims 1-13.

15. A composition comprising a VZV of any one of claims 1-13 or a vector of claim 14.

16. A method for treating a mammal having cancer, wherein said method comprises administering, to said mammal, a VZV of any one of claims 1-13, wherein the number of cancer cells within said mammal is reduced.

17. The method of claim 16, wherein said mammal is a human.

18. The method of any one of claims 16-17, wherein said cancer is selected from the group consisting of a melanoma, a glioma, a breast cancer, a T cell lymphoma, a neuroblastoma, a pancreatic cancer, a lung cancer, a liver cancer, a kidney cancer, a cervical cancer, a bladder cancer, and a prostate cancer.Attorney Docket No.07039-2319WO1 / 2024-024 19. The method of any one of claims 16-18, wherein said administering comprises intratumoral injection.

20. The method of any one of claims 16-19, wherein said method further comprises administering a cancer treatment to said mammal.

21. The method of claim 20, wherein said cancer treatment comprises administering an anti-cancer agent to said mammal.

22. The method of claim 20, wherein said cancer treatment comprises subjecting said mammal to surgery, a radiation therapy, or an adoptive cell transfer therapy.

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