Compositions comprising an oncolytic virus and a glial cell for use in treating neuroblastoma

Abstract

Described herein are methods and compositions for treating a disease, e.g. cancer (including solid tumors), using glial cells (e.g. microglia cells and / or astrocytes) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e.g., vaccinia virus). The methods and compositions provided herein demonstrate a synergistic anti-cancer effect.

Description

Atty Docket No. 055523-507001WGMETHODS AND COMPOSITIONS FOR TREATING NEUROBLASTOMARELATED APPLICATIONS

[0001] This patent application claims the benefit of U.S. Provisional Patent Application No. 63 / 708.892, filed on October 18, 2024, the contents of which are incorporated herein by reference for all purposes, including all text, tables and drawings.BACKGROUND

[0002] Microglia, the resident macrophages of the central nervous system (CNS), serve as the brain's first line of defense by engulfing harmful pathogens, including viruses, and clearing cellular debris, misfolded proteins, and dying cells to maintain homeostasis.

[0003] Neuroblastoma is the most prevalent cancer in infants and the third most common cancer in children after leukemia and brain tumors. There is a large unmet need for new, paradigm shifting therapies to address undruggable, untreatable, recurrent or treatment- nonresponsive cancers, despite the fact that many treatments currently exist and are used to successfully treat many types of cancer.

[0004] New methods of treating tumors of the nervous system (e.g., neuroblastoma, glioblastoma) are needed. The compositions and methods provided herein, inter alia, address these and other concerns in the art.BRIEF SUMMARY OF THE INVENTION

[0005] Described herein are compositions and methods for treating tumors that affect the nervous system (e.g., neuroblastoma, glioblastoma) in a subject in need thereof. Current regimens for treating such tumors may include chemotherapy, surgical resection, and / or radiotherapy. However, the infiltrative nature and anatomical location of these tumors present significant challenges for effective treatment. The field needs targeted therapeutics capable of efficiently killing tumor cells within the central nervous system (CNS) while minimizing off- target effects.

[0006] Vaccinia virus (VV) has been used in oncolytic virotherapy due to its ability to infect and lyse tumor cells. However, challenges to VV therapy include efficient delivery of the virus to tumor sites and shielding the virus from host immune clearance. In both primary brain tumors and brain metastases, endogenous microglia and CNS-associated macrophages are recruited to the tumor microenvironment and release metalloproteases, cytokines, and growth factors that may promote tumor invasion. Disrupting this pro-tumorigenic cycle may be useful for treatingAtty Docket No. 055523-507001WG tumors that affect the nervous system. Furthermore, it was believed that these immune cells in the CNS would be poor candidates for infection with oncolytic viruses such as VV due to the understanding that they would be poorly infected and / or not release the oncolytic virus. As a result, the use of immune cells in the CNS, such as glial cells, macrophages, etc., as potential cells in the treatment of such cancers was something that the skilled artisan would have avoided and would be counterintuitive. The embodiments herein reflect surprising and unexpected compositions and methods. These improvements are detailed herein

[0007] Thus, provided herein are improved compositions and methods for treating tumors affecting the nervous system, including by administering to the subject a composition including an oncolytic virus (e.g., vaccinia virus) and a glial cell (e.g., microglia). In embodiments, the glial cell is infected by the oncolytic virus prior to administration. Ideally, in at least some embodiments, the compositions and methods described herein can avoid one or more significant drawbacks of current approaches, including poor access to deep brain regions, inefficient viral delivery, and rapid immune clearance of the virus.

[0008] Some embodiments relate generally to compositions including a glial cell and an oncolytic virus. For example, the compositions include a microglial cell infected with a vaccinia virus. In some embodiments, the composition includes a CNS-associated macrophage infected with an oncolytic virus (e.g., vaccinia virus). In embodiments, the composition is formulated as a pharmaceutical composition further including a pharmaceutically acceptable excipient. In embodiments, the composition is administered to a subject in need thereof for the treatment of cancer.

[0009] In some embodiments, methods are provided for making a composition including a poxvirus and glial cells. In embodiments, the method includes obtaining glial cells and / or CNS- associated macrophages from a subject and contacting the cells with a poxvirus to form poxvirus- infected glial cells and / or poxvirus-infected CNS-associated macrophages. In embodiments, the glial cells are autologous, allogeneic, or xenogeneic. In embodiments, the contacting step is performed ex vivo, and the resulting composition is administered to the subject for therapeutic use.

[0010] In some embodiments, the use of glial cells as carriers or modulators of vaccinia virus provides unexpected advantages. For example, microglia may shield the virus from immune clearance, facilitate targeted delivery to tumor-associated regions, and enhance local immune activation. In embodiments, the combination of microglia and vaccinia virus results in synergistic anti-tumor effects, including increased tumor cell lysis, improved recruitment of effector immuneAtty Docket No. 055523-507001WD cells, and modulation of the tumor microenvironment (e.g., disruption of the pro-tumorigemc microenvironment).

[0011] Thus, in another aspect provided herein is a composition including an oncolytic virus and a glial cell. In embodiments, the glial cell is infected by the oncolytic virus. In embodiments, the glial cell is an oligodendrocyte, astrocyte, microglia cell, or ependymal cell. In embodiments, the glial cell is a microglia cell or an astrocyte.

[0012] In embodiments, the glial cell is derived from a subject to be treated with the composition. In embodiments, the glial cell is allogeneic to a subject to be treated with the composition.

[0013] In embodiments, the glial cell is genetically modified. In embodiments, the glial cell is genetically modified to target a cancer cell.

[0014] In embodiments, the virus does not lyse the glial cell for at least 5 days after infection.

[0015] In embodiments, the composition includes a plurality of glial cells. In embodiments, the plurality of glial cells is infected by the oncolytic virus. In embodiments, the plurality of glial cells includes oligodendrocytes, astrocytes, microglia cells, ependymal cells, or a mixture thereof.

[0016] In embodiments, the oncolytic virus is a poxvirus. In embodiments, the poxvirus is a vaccinia virus. In embodiments, the vaccinia virus is selected from Dryvax, AC AMI 000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve. Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J, and IHD-W, Brighton, Dairen I and Connaught strains. In embodiments, the vaccinia virus is a Lister strain. In embodiments, the vaccinia virus is a Copenhagen strain.

[0017] In another aspect is provided a pharmaceutical composition including the composition described herein and a pharmaceutically acceptable excipient.

[0018] In another aspect is provided a method for treating cancer in a subject in need thereof, the method including administering to the subject a composition as provided herein. In embodiments, the subject is a human.

[0019] In embodiments, the cancer is neuroblastoma.

[0020] In embodiments, the composition is administered to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra-parenchymal,Atty Docket No. 055523-507001WG intraventricular, intraarticular, or intraocular injection. In embodiments, the composition is administered directly to a region affected by the disease. In embodiments, the composition is administered by MRI-guided delivery.

[0021] In another aspect is provided a method for making a composition as described herein. In embodiments, the method includes: (a) obtaining glial cells; and (b) contacting the glial cells with a poxvirus to form poxvirus-infected glial cells.

[0022] In embodiments, the glial cells are isolated prior to step b). In embodiments, the glial cells are obtained from a subject to be treated with the composition. In embodiments, the glial cells are derived in vitro, for example from embryonic stem cells, pluripotent stem cells, induced pluripotent stem cells, or a precursor of glial cells. In embodiments, the glial cells are allogeneic to a subject to be treated with the composition.

[0023] In embodiments, the glial cells are infected with the poxvirus at a titer > 0.5 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1 MOI.

[0024] In embodiments, the glial cells are incubated with the virus for about 1 day to about 5 days. In embodiments, the glial cells are incubated with the virus for about 3 days.

[0025] In embodiments, the poxvirus does not lyse the glial cells.

[0026] In another aspect is provided a use of a composition provided herein including embodiments thereof in a method of treating cancer in a subject in need thereof, the method including administering to the subject the composition provided herein including embodiments thereof.

[0027] In another aspect is provided a use of a composition provided herein including embodiments thereof in the preparation of a medicament for treating cancer in a subject in need thereof, the method including administering to the subject the composition provided herein including embodiments thereof.

[0028] In embodiments, the cancer is neuroblastoma or glioma.

[0029] In embodiments, the composition is administered to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra-parenchymal, intraventricular, intraarticular, or intraocular injection.

[0030] In embodiments, the composition is administered directly to a region affected by the disease. In embodiments, the composition is administered by MRI-guided delivery.Atty Docket No. 055523-507001WG

[0031] In embodiments, the subject is a human.BRIEF DESCRIPTION OF THE FIGURES

[0032] FIG. 1 shows IncuCyte results, demonstrating that BV2 cells can be infected with different MOIs of VV. According to the fluorescence intensity over time, the highest infection rate occurs within the first 48 hours. During this period, most of the BV2 cells at MOI 1 and MOI 10 show significant cell death.

[0033] FIG. 2 shows MTT assay results showing that an MOI of 0.1 of VV used to infect BV2 cells is non-toxic and does not affect cell viability. However, MOIs of 1 and 10 significantly reduced cell viability, with nearly 50% of cells affected after 48 hours.

[0034] FIG. 3 shows plaque assay results showing that BV2 cells infected with VV at different time points post-infection successfully release the virus. At MOIs of 1 and 10, there is significantly higher viral release compared to MOI 0.1, which demonstrates minimal viral production.

[0035] FIGs. 4A-4B show SK-N-AS-GFP cells treated with varying ratios of uninfected BV2 cells (FIG. 4A) and infected BV2 cells (48 hours post-infection; FIG. 4B). Additionally, vaccinia virus (VV) at MOI 1 was applied. The treatments were monitored using IncuCyte. The data show that uninfected BV2 cells did not reduce cancer cell viability. However, BV2 cells infected with MOI 1 at ratios of 1 : 1 and 5: 1 significantly reduced cancer cell viability after 72 hours. VV began infecting and killing the cancer cells by 96 hours.

[0036] FIGs. 5A-5B show different ratios of uninfected BV2 cells (FIG. 5A), BV2 cells infected with MOI 1 (48 hours post-infection), and VV (MOI 1; FIG. 5B) co-cultured with SH- SY5Y-GFP cells. Data show that uninfected BV2 cells do not affect the viability of cancer cells. However, infected BV2 cells, at ratios of 1 : 1 and 5: 1, successfully eliminate cancer cells after 72 hours.

[0037] FIG. 6 shows a graph displaying fluorescence intensity over time (based on FIG. 5) revealing that in uninfected treatments, fluorescence increases, indicating cancer cell proliferation. In contrast, fluorescence decreases in infected treatments, signifying cancer cell death.

[0038] FIGs. 7A-7D show SH-SY5Y cells and co-cultured cancer cells with BV2 microglial cells (both infected and uninfected) cultured in a 3D model using the hanging drop method.Atty Docket No. 055523-507001WOUnder light microscopy at 4X magnification, the 3D structures were observed (FIG. 7A). FIG. 7B: SH-SY5Y cells with or without VV. FIG. 7C: BV2 cells with or without VV. FIG. 7B: SH- SY5Y cells and BV2 cells, with or without VV. IncuCyte data revealed that SH-SY5Y cells exhibited continuous growth over time. However, in coculture with infected BV2 cells, SH- SY5Y cells were eliminated, indicating a cytotoxic effect.

[0039] FIGs. 8A-8C show FIG. 8A: IncuCyte images showing BV2 cell infection by VV at MOIs of 0.1, 1, and 10. FIG. 8B: MTT assay results demonstrate that MOIs of 1 and 10 significantly reduced BV2 cell viability at 48 hours post-infection. FIG. 8C: Plaque assay showing time-dependent viral release from BV2 cells infected with VV. **** p < 0.0001; *** p < 0.005.

[0040] FIGs. 9A-9B show FIG. 9A: IncuCyte images of SH-SY5Y cells co-cultured with VV- infected (MOI 1) or non-infected BV2 cells. FIG. 9B: IncuCyte images of U87 cells co-cultured with VV-infected (MOI 1) or non-infected BV2 cells.

[0041] FIGs. 10A-10B show FIG. 10A: MTT assay showing that VV-infected BV2 cells significantly reduced SH-SY5Y (top panel) and U87 (bottom panel) cancer cell viability at 72 hours post-co-culture. FIG. 10B: Plaque assay demonstrating that VV-infected BV2 cells co- cultured with SH-SY5Y (top panel) and U87 (bottom panel) cancer cells released infectious virus at multiple time points. *** p < 0.005; **** p < 0.0001.

[0042] FIG. 11 shows fluorescence microscopy images of cancer cells in the upper chamber after 96 hours, comparing conditions with non-infected BV2 cells, VV-infected BV2 cells, and 10% FBS with free VV.

[0043] FIGs. 12A-12B show Growth kinetics of SH-SY5Y (FIG. 12A) and U87 (FIG. 12B) tumor spheroids. Images were captured daily using the IncuCyte system, and spheroid diameters were measured at 4* magnification.

[0044] FIG. 13 shows SH-SY5Y 3D spheroid models co-cultured with VV-infected or noninfected BV2 cells or directly treated with VV (MOI 1). Images were captured with the IncuCyte system; sectioned spheroids were visualized by fluorescence microscopy.

[0045] FIG. 14 shows U87 3D spheroid models co-cultured with VV-infected or non-infected BV2 cells ordirectly treated with VV (MOI 1). Images were captured with the IncuCyte system; sectioned spheroids were visualized by fluorescence microscopy.Atty Docket No. 055523-507001WDDETAILED DESCRIPTION

[0046] After reading this description it will become apparent to one skilled in the art how to implement the invention in various alternative embodiments and alternative applications. However, all the various embodiments of the present invention will not be described herein. It will be understood that the embodiments presented here are presented by way of an example only, and not limitation. As such, this detailed description of various alternative embodiments should not be construed to limit the scope or breadth of the present invention as set forth below.

[0047] Before the present invention is disclosed and described, it is to be understood that the aspects described below are not limited to specific compositions, methods of preparing such compositions, or uses thereof as such may, of course, vary, ft is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting.

[0048] The detailed description of the invention is divided into various sections only for the reader’s convenience and disclosure found in any section may be combined with that in another section. Titles or subtitles may be used in the specification for the convenience of a reader, which are not intended to influence the scope of the present invention.I. DEFINITIONS

[0049] Unless defined otherwise, 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 belongs. In this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings:

[0050] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0051] “Optional” or “optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0052] The term “about” when used before a numerical designation, e.g. , temperature, time, amount, concentration, and such other, including a range, indicates approximations which mayAtty Docket No. 055523-507001WG vary by ( + ) or ( - ) 10%, 5%,1%, or any subrange or subvalue there between. Preferably, the term ‘‘about” when used with regard to a dose amount means that the dose may vary by + / - 10%.

[0053] “Comprising” or “comprises” is intended to mean that the compositions and methods include the recited elements, but not excluding others. “Consisting essentially of’ when used to define compositions and methods, shall mean excluding other elements of any essential significance to the combination for the stated purpose. Thus, a composition consisting essentially of the elements as defined herein would not exclude other materials or steps that do not materially affect the basic and novel characteristic(s) of the claimed invention. “Consisting of ’ shall mean excluding more than trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

[0054] The terms “disease” or “condition” refer to a state of being or health status of a patient or subject capable of being treated with the compounds or methods provided herein. The disease may be a cancer, such as neuroblastoma.

[0055] The terms “treating”, or “treatment” refers to any indicia of success in the therapy or amelioration of an injury, disease, pathology' or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving a patient’s physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination, neuropsychiatric exams, and / or a psychiatric evaluation. The term "treating" and conjugations thereof, may include prevention of an injury, pathology, condition, or disease. In embodiments, treating is preventing. In embodiments, treating does not include preventing.

[0056] “Treating” or “treatment” as used herein (and as w ell-understood in the art) also broadly includes any approach for obtaining beneficial or desired results in a subject's condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to. alleviation or amelioration of one or more symptoms or conditions, diminishment of the extent of a disease, stabilizing (z.e., not w orsening) the state of disease, prevention of a disease’s transmission or spread, delay or slow ing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable. In other words, "treatment" as used herein includes any cure, amelioration, or prevention of a disease. Treatment may7prevent the disease from occurring;Atty Docket No. 055523-507001WG inhibit the disease’s spread; relieve the disease’s symptoms, fully or partially remove the disease’s underlying cause, shorten a disease’s duration, or do a combination of these things.

[0057] "Treating" and "treatment" as used herein include prophylactic treatment. Treatment methods include administering to a subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may include a series of administrations. The length of the treatment period depends on a variety of factors, such as the severity of the condition, the age of the patient, the concentration of active agent, the activity' of the compositions used in the treatment, or a combination thereof. It will also be appreciated that the effective dosage of an agent used for the treatment or prophylaxis may increase or decrease over the course of a particular treatment or prophylaxis regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. For example, the compositions are administered to the subject in an amount and for a duration sufficient to treat the patient. In embodiments, the treating or treatment is not prophylactic treatment.

[0058] The term “prevent” refers to a decrease in the occurrence (or recurrence) of disease symptoms in a patient. As indicated above, the prevention may be complete (no detectable symptoms) or partial, such that fewer symptoms are observed than would likely occur absent treatment.

[0059] “Patient,” “subject,” or “subject in need thereof’ refers to a living organism suffering from or prone to a disease or condition that can be treated by administration of a pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goat, sheep, pigs, deer, and other non-mammalian animals. In some embodiments, a patient is human. In embodiments, the human is a pediatric patient. In embodiments, a patient is a domesticated animal (e.g., goat, sheep, cow, horse, etc.). In embodiments, a patient is a companion animal, including but not limited to canine, feline, rodent (mouse, rat, gerbil, hamster, guinea pig, chinchilla, and the like), rabbit, ferret, etc.

[0060] An “effective amount” is an amount sufficient for a compound to accomplish a stated purpose relative to the absence of the compound (e.g. achieve the effect for which it is administered, treat a disease, or reduce one or more symptoms of a disease or condition). An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.” A “reduction” of a symptom or symptoms (and grammatical equivalents of this phrase) means decreasing of the severity or frequency of theAtty Docket No. 055523-507001WD symptom(s), or elimination of the symptom(s). The exact amounts will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology’ of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy. 20th Edition. 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0061] As is well known in the art, therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a dose that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring effectiveness and adjusting the dosage upwards or downwards, as described herein. Adjusting the dose to achieve maximal efficacy in humans based on the methods described herein and other methods is well within the capabilities of the ordinarily skilled artisan.

[0062] The term “therapeutically effective amount,'’ as used herein, refers to that amount of the therapeutic agent sufficient to ameliorate the disorder, as described above. For example, for the given parameter, a therapeutically effective amount w ill show' an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy can also be expressed as “-fold"’ increase or decrease. For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more effect over a control.

[0063] Dosages may be varied depending upon the requirements of the patient and the composition being employed. The dose administered to a patient, in the context of the present disclosure, should be sufficient to effect a beneficial therapeutic response in the patient over time. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects. Determination of the proper dosage for a particular situation is within the skill of the practitioner. In some embodiments, treatment is initiated with smaller dosages which are less than the optimum dose of the composition. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached. Dosage amounts and intervals can be adjusted individually to provide levels of the administered composition effective for the particular clinical indication being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual's disease state.

[0064] As used herein, the term "administering" means oral administration, administration as a suppository', topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional,Atty Docket No. 055523-507001WG intrathecal, intra-cerebro- ventricular, intrapleural, intra-parenchymal, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, etc. Administration also includes direct administration, e.g., directly to a site of inflammation. Direct administration maybe via guided delivery, e g., magnetic resonance imaging (MRI)-guided delivery. In embodiments, the administering does not include administration of any active agent other than the recited active agent.

[0065] "Co-administer" is meant that a composition described herein is administered at the same time, just prior to, or just after the administration of one or more additional therapies. The compositions provided herein can be administered alone or can be co-administered to the patient. Co-administration is meant to include simultaneous or sequential administration of the compositions individually or in combination (more than one composition). Thus, the preparations can also be combined, when desired, with other active substances.

[0066] The term “autologous,"’ “autologous cell” or “autologous transplantation” as used herein in relation to cell transplantation indicates that the donor and recipient of the cells is the same individual. The term “allogenic,” “allogenic cell” or “allogenic transplantation” as used herein in relation to cell transplantation indicates that the donor and recipient of the cells are different individuals of the same species.

[0067] The terms “glial cells,” “neuroglia,” and the like are used herein according to their plain and ordinary meaning. In some embodiments, glial cells refer to cells that support nerve cells. In embodiments, glial cells maintain the ionic milieu of nerve cells. In embodiments, glial cells modulate the rate of nerve signal propagation. In embodiments, glial cells modulate synaptic action by controlling the uptake, recycling, or degradation of neurotransmitters. In embodiments, glial cells provide a scaffold for some aspects of neural development, including guidance of neuronal migration and axonal pathfinding. In embodiments, glial cells aid in, or prevent, recovery from neural injury- by participating in tissue remodeling, debris clearance, and modulation of inflammatory responses. In embodiments, glial cells regulate the formation and maintenance of the blood-brain barrier. In embodiments, glial cells contribute to cerebrospinal fluid production, circulation, and homeostasis. In embodiments, glial cells participate in immuneAtty Docket No. 055523-507001WG surveillance and neuroimmune signaling within the central nervous system. In embodiments, glial cells influence synaptic plasticity and contribute to long-term changes in neural circuitry. In embodiments, glial cells include oligodendrocytes, astrocytes, microglia, and / or ependymal cells. In embodiments, glial cells include microglia-like cells derived from human monocytes.

[0068] The terms “microglia,” “microglia cells,” “microglial cells,” and the like are used herein according to their plain and ordinary' meaning. In some embodiments, microglia refer to glial cells that function as resident immune cells of the central nervous system (CNS). In embodiments, microglia are located within the brain parenchyma. In embodiments, microglia patrol the CNS to detect and respond to pathogens, cellular damage, and other perturbations in the neural environment. In embodiments, microglia remove cellular debris from sites of injury or normal cell turnover through phagocytosis. In embodiments, microglia modulate neuroinflammatory responses by releasing cytokines, chemokines, and other signaling molecules. In embodiments, microglia contribute to synaptic remodeling and plasticity by pruning neuronal connections during development and in response to neural activity. In embodiments, microglia interact with neurons, astrocytes, and other glial cells to maintain CNS homeostasis. In embodiments, microglia are derived from yolk sac progenitors during embryonic development. In embodiments, microglia are long-lived and capable of self-renewal within the CNS. In embodiments, microglia express molecular markers that distinguish them from other CNS- associated immune cells, including TMEM119, P2RY12, and / or Sall 1. In embodiments, the microglial cell is a mouse microglial cell. In embodiments, the microglial cell is a BV2 cell. In embodiments, the microglial cell is a human microglial cell. In embodiments, the microglial cell is a HMC3 cell. In embodiments, the microglial cell is a human monocyte-derived microglia-like cell. In embodiments, the human monocyte-derived microglia-like cell expresses one or more microglial markers, including TMEM119, P2RY12, and / or IBA1. Eluman monocyte-derived microglial -like cells are well known in the art (See Ryan et al., Sei Trans I Med, 2017, 9, 421, eaai7635, pp. 1-12. which is incorporated herein by reference in its entirety and for all purposes).

[0069] The terms “astrocytes” and the like are used herein according to their plain and ordinary meaning. In some embodiments, astrocytes refer to glial cells restricted to the brain and spinal cord that possess elaborate local processes. In embodiments, astrocytes maintain an appropriate chemical environment for neuronal signaling through regulation of extracellular ion concentrations and neurotransmitter clearance. In embodiments, astrocytes contribute to the formation, maintenance, and regulation of the blood-brain barrier. In embodiments, astrocytes modulate synaptic transmission and plasticity by releasing gliotransmitters and responding toAtty Docket No. 055523-507001WG neuronal activity. In embodiments, astrocytes provide metabolic support to neurons, including the uptake and redistribution of glucose and lactate. In embodiments, astrocytes participate in the repair and scarring processes following CNS injury. In embodiments, astrocytes influence neurovascular coupling and regulate cerebral blood flow. In embodiments, astrocytes interact with microglia, oligodendrocytes, neurons, and endothelial cells to maintain CNS homeostasis. In embodiments, astrocytes are derived from neuroectodermal progenitors during development. In embodiments, astrocytes are capable of limited proliferation and may undergo reactive changes in response to CNS pathology. In embodiments, astrocytes express molecular markers including GFAP (glial fibrillary acidic protein), S 1OO0, and / or ALDH1L1.

[0070] “The terms “oligodendrocytes7’ and the like are used herein according to their plain and ordinary meaning. In some embodiments, oligodendrocytes refer to glial cells restricted to the central nervous system (CNS) that form myelin sheaths around axons. In embodiments, oligodendrocytes wrap their membrane processes around neuronal axons to create compact myelin, which facilitates rapid saltatory conduction of action potentials. In embodiments, oligodendrocytes provide metabolic and trophic support to axons, including the transfer of energy substrates such as lactate. In embodiments, oligodendrocytes contribute to the structural organization of w hite matter tracts in the brain and spinal cord. In embodiments, oligodendrocytes arise from oligodendrocyte precursor cells (OPCs), which differentiate during development and in response to injury. In embodiments, oligodendrocytes are capable of limited regeneration and remyelination following demyelinating injury. In embodiments, oligodendrocytes express molecular markers including myelin basic protein (MBP), proteolipid protein (PLP), and / or oligodendrocyte transcription factor 2 (OLIG2). In embodiments, oligodendrocytes interact with astrocytes, microglia, and neurons to maintain CNS homeostasis and support neural function.

[0071] The terms “ependymal cells” and the like are used herein according to their plain and ordinary meaning. In some embodiments, ependymal cells refer to glial cells that line the ventricular system of the brain and the central canal of the spinal cord. In embodiments, ependymal cells form a barrier between the cerebrospinal fluid (CSF) and the neural tissue of the central nervous system (CNS). In embodiments, ependymal cells contribute to the production, circulation, and regulation of CSF. In embodiments, ependymal cells possess motile cilia that facilitate the movement of CSF through the ventricular system. In embodiments, ependymal cells participate in the exchange of signaling molecules between the CSF and the brain parenchyma. In embodiments, ependymal cells may serve as a source of neural stem or progenitor cells in certain regions of the CNS. In embodiments, ependymal cells are derived from neuroectodermalAtty Docket No. 055523-507001WG progenitors during development. In embodiments, ependymal cells express molecular markers including vimentin, SI 00(3, and / or CD24. In embodiments, oligodendrocytes are distinguished from other glial cell ty pes by their unique morphology7and ventricular localization.

[0072] The terms "macrophage." “macrophages,” and the like are used herein according to their plain and ordinary7meaning. In some embodiments, macrophages refer to immune cells that reside at anatomical interfaces of the central nervous system (CNS), including the meninges, perivascular spaces, and / or choroid plexus. In embodiments, macrophages contribute to immune surveillance and barrier integrity7at CNS borders. In embodiments, macrophages respond to infection, injury, or inflammation by releasing cytokines, presenting antigens, and recruiting peripheral immune cells. In embodiments, macrophages regulate cerebrospinal fluid composition and participate in clearance of waste products and signaling molecules. In embodiments, macrophages are derived from bone marrow-derived monocytes and are replenished from the peripheral circulation. In embodiments, macrophages are distinct from microglia in their developmental origin, anatomical localization, and molecular profile. In embodiments, the macrophage is a CNS-associated macrophage. In embodiments, macrophages express molecular markers including CD45hlgh, CD1 lb, and / or F4 / 80. In embodiments, macrophages do not typically migrate into the brain parenchyma under homeostatic conditions but may infiltrate the parenchyma during pathological states.

[0073] An “oncolytic virus” is a virus that can infect and kill cancer cells. Examples of oncolytic viruses include, but are not limited to, adenoviruses, herpes viruses, measles viruses, coxsackie viruses, polioviruses, reoviruses, poxviruses, vaccinia viruses, Vesicular stomatitits virus, senecavirus, RIGVIR, semliki forrest virus, maraba virus and Newcastle disease viruses.

[0074] In embodiments, the oncolytic virus (e.g., a poxvirus or a vaccinia virus) and the glial cell (e.g.. microglia) and / or macrophage (e.g., CNS-associated macrophage) are present in a combined synergistic amount, wherein the combined synergistic amount is effective to treat cancer in a subject in need thereof. A “combined synergistic amount” is used herein according to its plain and ordinary meaning. In embodiments, combined synergistic amount refers generally to the sum of a first amount (e.g.. an amount of oncolytic virus) and a second amount (e.g., an amount of a glial cell) that results in a synergistic effect (i.e. an effect greater than an additive effect). Therefore, the terms "synergy," "synergism," "synergistic," which are used herein interchangeably, refer to a measured effect of compounds (e.g., oncolytic virus and a glial cell) administered in combination where the measured effect (e.g., treatment of cancer) is greater than the sum of the individual effects of each of the compounds administered alone as a single agent.Atty Docket No. 055523-507001WDIn embodiments, the combined synergistic amount is a combined therapeutically effective amount for treating cancer in a subject in need thereof. In embodiments, the combined synergistic amount includes an amount of the oncolytic virus (e.g., poxvirus or vaccinia virus) that is less than a therapeutically effective amount of the oncolytic virus when administered alone as a single agent. In embodiments, the combined synergistic amount includes an amount of a glial cell (e.g. microglia) and / or macrophage (e.g., CNS-associated macrophage) that is less than a therapeutically effective amount of the glial cell and / or macrophage when administered alone as a single agent. In embodiments, the combined synergistic amount includes an amount of oncolytic virus and an amount of a glial cell and / or macrophage that are each less than a therapeutically effective amount of the oncolytic virus and the therapeutically effective amount of the glial cell and / or macrophage when each of the compounds (e.g., oncolytic virus and the glial cell and / or macrophage) are administered alone as a single agent. In embodiments, the therapeutically effective amount of the oncolytic virus (e.g., poxvirus or vaccinia virus) is an U.S. Food and Drug Administration (FDA)-approved amount of oncolytic virus that is effective in treating a disease, disorder, or condition in a subject in need thereof. In embodiments, the therapeutically effective amount of a glial cell and / or macrophage is an FDA-approved amount of a glial cell and / or macrophage that is effective in treating a disease, disorder, or condition in a subject in need thereof. In embodiments, the combined therapeutically effective amount is a therapeutically effective amount of oncolytic virus combined with a therapeutically effective amount of a glial cell and / or macrophage. In embodiments, the synergistic effect may be a synergistic anti-cancer effect.

[0075] In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6. 1.7, 1.8, 1.9, 2.0. 2.1, 2.2, 2.3, 2.4. 2.5, 2.6, 2.7, 2.8. 2.9, 3.0,3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2,5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4,7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6,9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17. 18. 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32. 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43. 44. 45. 46. 47. 48. 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the oncolytic virus when used separately from the glial cell and / or macrophage. In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6. 0.7, 0.8, 0.9, 1.0. 1.1, 1.2, 1.3, 1.4. 1.5, 1.6, 1.7,1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1,Atty Docket No. 055523-507001WD6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3,8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54. 55. 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80. 81. 82. 83. 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the glial cell and / or macrophage when used separately from the oncolytic virus.

[0076] In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0,3.1, 3.2, 3.3, 3.4. 3.5, 3.6, 3.7, 3.8. 3.9, 4.0, 4.1, 4.2. 4.3, 4.4, 4.5, 4.6. 4.7, 4.8, 4.9, 5.0. 5.1, 5.2,5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4,7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6,9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43. 44. 45. 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the oncolytic virus when used separately from the microglia. In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4. 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1,2.2, 2.3, 2.4, 2.5. 2.6, 2.7, 2.8, 2.9. 3.0, 3.1, 3.2, 3.3. 3.4, 3.5, 3.6, 3.7. 3.8, 3.9, 4.0, 4.1. 4.2, 4.3,4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5,6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7,8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32. 33. 34. 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the microglia when used separately from the oncolytic virus.

[0077] The term "Cluster of Differentiation protein" or "CD protein" or “cluster of designation” or “classification determinant” as used herein includes any of the cell surface proteins identified as targets for immunophenotyping cells. CD proteins can act as receptors or ligands for participation in immune responses. CD proteins include, but are not limited to, CD3, CD4, CD5, CD7, CD8, CD19, CD20, CD22, CD30, CD33, CD44v6, CD123, CD138, CD171, CD4, CD8 or CD45RA+.Atty Docket No. 055523-507001WD

[0078] The term “vaccine’7as used herein, refers to any type of biological preparation contributing to or soliciting active immune responses against a particular disease or pathogen. Such biological preparation can include, but is not limited to, an antigen derived from a diseasecausing agent or a portion of an antigen derived from a disease-causing agent. Such biological preparation can also be in the form of live attenuated preparation, including live, or weakened or modified disease causing agents or pathogens; or in the form of inactivated or killed diseasecausing agents or pathogens. Alternative forms of such biological preparation further include, but are not limited to, the forms of subunit, toxoid, conjugate, DNA and recombinant vectors, or any suitable forms that might become developed or available in the future for soliciting active immune responses there against.

[0079] It should be noted that in some embodiments, while the term “vaccine” is used herein, the vaccine need not provide significant immunity against smallpox (or any other pathogen), so long as it is effective against a disease as described herein. For example, the vaccine may be any immunogenic or infectious composition that treats the disease. In some cases, the term is used to identity- certain materials or compositions, and not necessarily the ability of material or composition to provide immunity against smallpox, for example. The virus can be from any strain of virus, including for example, one or more listed below and elsewhere herein, including those that are not part of approved or contemplated vaccines.

[0080] As used herein, “virus" refers to any of a large group of entities referred to as viruses. Viruses typically contain a protein coat surrounding an RNA or DNA core of genetic matenal, but no semipermeable membrane, and are capable of growth and multiplication only in living cells. Viruses for use in the methods provided herein include, but are not limited, to a poxvirus, adenovirus, herpes simplex virus, Newcastle disease virus, vesicular stomatitis virus, mumps virus, influenza virus, measles virus, reovirus. human immunodeficiency virus (HIV), hanta virus, myxoma virus, cytomegalovirus (CMV), lentivirus, and any plant or insect virus. In particular, a virus as used herein is an oncolytic virus.

[0081] As used herein, “heterologous nucleic acid” refers to a nucleic acid, DNA or RNA, which has been introduced into a virus or a cell (or the cell's ancestor). Such heterologous nucleic acid may comprise the sequence and operable regulatory elements for genes. For example, the heterologous nucleic acid may comprise a selection marker gene, a suicide gene, or a gene expressing a useful protein product that is not expressed endogenously, or expressed endogenously at low levels.Atty Docket No. 055523-507001WG

[0082] As used herein, the term "concurrently” as referring to administration of an oncolytic virus and a cell, refers to administration within 48 hours of each other. In some embodiments, the oncolytic virus and cell are administered within 36 hours of each other, within 24 hours of each other, within 12 hours of each other, within 10 hours of each other, within 8 hours of each other, within 6 hours of each other, within 4 hours of each other, within two hours of each other, within 1 hour of each other.

[0083] The term “autologous,” “autologous cell” or “autologous transplantation” as used herein in relation to cell transplantation indicates that the donor and recipient of the cells is the same individual. The term “allogenic,” “allogenic cell” or “allogenic transplantation” as used herein in relation to cell transplantation indicates that the donor and recipient of the cells are different individuals of the same species.

[0084] “Pharmaceutically acceptable excipient” and “pharmaceutically acceptable carrier” refer to a substance that aids the administration of an active agent to and / or absorption by a subject and can be included in the compositions of the present disclosure without causing a significant adverse toxicological effect on the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCI, normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethy cellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure. One of skill in the art will recognize that other pharmaceutical excipients are useful in the present disclosure.II. ONCOLYTIC VIRUSES

[0085] Oncolytic viruses are viruses that target cancer cells. Oncolytic viruses can be genetically engineered to selectively replicate in and lyse cancer cells, while avoiding healthy cells. Oncolytic viruses destroy cancer cells by either: 1) directly infecting a cancer cell, and lysing it, or 2) recruiting and directing a refreshed host immune response to the cancer cells.

[0086] The potential for oncolytic viruses as a treatment for cancer was initially discovered after a patient with myelogenous leukemia was infected with influenza, and went into spontaneous remission. Viruses from many classes have been genetically engineered to be oncolytic, including, but are not limited to: adenoviruses, herpes viruses, measles viruses,Atty Docket No. 055523-507001WG coxsackie viruses, polioviruses, reoviruses, poxviruses, vaccinia viruses, Vesicular stomatitits virus, senecavirus, RIGVIR, semliki forrest virus, maraba virus and Newcastle disease viruses. Viral components are the targets for genetic engineering, and include: adding tumor-specific promoters, viral gene knockouts, viral capsid modification, and expression of immune systemactivating agents including antibodies, cytokines, and costimulatory molecules.

[0087] Variola virus is the cause of smallpox. Unlike variola virus, vaccinia virus does not normally cause systemic disease in immune-competent individuals and it has therefore been used as a live vaccine to immunize against smallpox. Smallpox has been eradicated as a natural disease due to successful worldwide vaccination with vaccinia virus. Routine smallpox vaccination has been discontinued for many years, except for people at higher risk of poxvirus infections (e.g., laboratory workers). Although the United States discontinued routine childhood immunization against smallpox in 1972, the use of smallpox vaccine is generally considered safe for pediatric use.

[0088] Attenuated strains derived from a pathogenic virus can be used for the manufacturing of a live vaccine. Non-limiting examples of viral strains that have been used as a smallpox vaccine include but are not limited to the Lister (also known as Elstree), New York City Board of Health C'NYCBH strain”), Dairen, Ikeda, LC16M8, Western Reserve (WR), Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J, and IHD-W, Brighton, Ankara, MV A, Dairen I, LIPV, LC16MO, LIVP, WR 65-16, EM63, and Connaught strains. In some embodiments, the smallpox vaccine as disclosed herein is an attenuated New York City Board of Health (NYCBOH) strain of vaccinia virus. In some embodiments, the NYCBOH strain of vaccinia virus may be ATCC VR- 118 or CJ-MVB-SPX.

[0089] Surprisingly, an oncolytic virus can infect microglia cells and / or astrocytes, and the infected microglia cells or astrocytes can be used to target and kill tumor cells, including neuroblastoma. In embodiments, the oncolytic virus is a poxvirus.

[0090] In embodiments, the poxvirus is vaccinia virus. In some embodiments, the vaccinia virus is selected from Dryvax, AC AMI 000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen. Ikeda, LC16M8. Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I and Connaught strains. In some embodiments, the vaccinia virus strain is Drayvax. In some embodiments, the vaccinia virus strain is AC AMI 000. In some embodiments, the vaccinia virus strain is ACAM2000. In some embodiments, the vaccinia virus strain is Lister. In some embodiments, the vaccinia virus strain is EM63. In some embodiments, the vaccinia virus strainAtty Docket No. 055523-507001WD is LIVP. In some embodiments, the vaccinia virus strain is Tian Tan. In some embodiments, the vaccinia virus strain is Copenhagen. In some embodiments, the vaccinia virus strain is Western Reserve. In some embodiments, the vaccinia virus strain is Modified Vaccinia Ankara (MV A). In some embodiments, the vaccinia virus strain is Ikeda. In some embodiments, the vaccinia virus strain is New York City Board of Health. In some embodiments, the vaccinia virus strain is Dairen. In some embodiments, the vaccinia virus strain is LC16M8. In some embodiments, the vaccinia virus strain is Tashkent. In some embodiments, the vaccinia virus strain is Wyeth. In some embodiments, the vaccinia virus strain is IHD-J. In some embodiments, the vaccinia virus strain is IHD-W. In some embodiments, the vaccinia virus strain is Brighton. In some embodiments, the vaccinia virus strain is Dairen I. In some embodiments, the vaccinia virus strain is Connaught.

[0091] In embodiments, the poxvirus is oncolytic. In some embodiments, the vaccinia virus is oncolytic. In some embodiments, the oncolytic vaccinia virus selected from Dryvax, ACAM1000. ACAM2000. Lister, EM63, LIVP. Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I and Connaught strains. In some embodiments, vaccinia virus strain Dryvax is oncolytic. In some embodiments, vaccinia virus strain Dry vax is AC AMI 000. In some embodiments, vaccinia virus strain ACAM2000 is oncolytic. In some embodiments, vaccinia virus strain Lister is oncolytic. In some embodiments, vaccinia virus strain EM63 is oncolytic. In some embodiments, vaccinia virus strain LIVP is oncolytic. In some embodiments, vaccinia virus strain Tian Tan is oncolytic. In some embodiments, vaccinia virus strain Copenhagen is oncolytic. In some embodiments, vaccinia virus strain Western Reserve is oncolytic. In some embodiments, vaccinia virus strain Modified Vaccinia Ankara (MV A) is oncolytic. In some embodiments, vaccinia virus strain New York City Board of Health is oncolytic. In some embodiments, vaccinia virus strain Dairen is oncolytic. In some embodiments, vaccinia virus strain Ikeda is oncolytic. In some embodiments, vaccinia virus strain LC16M8 is oncolytic. In some embodiments, vaccinia virus strain Tashkent is oncolytic. In some embodiments, vaccinia virus strain Wyeth is oncolytic. In some embodiments, vaccinia virus strain IHD-J is oncolytic. In some embodiments, vaccinia virus strain IHD-W is oncolytic. In some embodiments, vaccinia virus strain Brighton is oncolytic. In some embodiments, vaccinia virus strain Dairen I is oncolytic. In some embodiments, vaccinia virus strain Connaught is oncolytic.Atty Docket No. 055523-507001WG

[0092] In embodiments, the vaccinia virus does not include the Lister strain vaccinia virus. In embodiments, the vaccinia virus does not include the Lister vaccinia virus. In embodiments, the vaccinia virus does not include the LIVP 1.1.1 vaccinia virus.

[0093] In embodiments, the oncolytic virus does not lyse a microglia cell. In some embodiments, the oncolytic virus does not lyse an allogenic microglia cell. In some embodiments, the oncolytic virus does not lyse an autologous microglia cell. In some embodiments, the oncolytic virus does not lyse an astrocyte. In some embodiments, the oncolytic virus does not lyse an allogenic astrocyte. In some embodiments, the oncolytic virus does not lyse an autologous astrocyte. In embodiments, “does not lyse'’ indicates that the virus does not lyse the cell for at least 12 hours, 24 hours, 36 hours, 48 hours, 60 hours, 72 hours. 86 hours. 98 hours or more (or any sub value or subrange therein including endpoints) after infection of the cell with the virus.

[0094] U.S. Patent No. 10,105,436, which is incorporated herein by reference in its entirety for all that is taught therein, describes poxviruses, including smallpox vaccines, that can be used in the methods and compositions described herein.III. GLIAL COMPOSITIONS

[0095] Any method for making (e.g., deriving, differentiating, isolating, culturing, expanding, etc.) and using glial cells (e.g., microglia) may be used with the compositions and methods described herein. Methods of making and using microglia cells are well known in the art.

[0096] In an aspect, provided herein, is a composition containing an oncolytic virus and a glial cell, wherein the cell is infected by the oncolytic virus.

[0097] In embodiments, the glial cell is an oligodendrocyte, astrocyte, microglia, or ependymal cell. In embodiments, the glial cell is an oligodendrocyte. In embodiments, the glial cell is an astrocyte. In embodiments, the glial cell is a microglia. In embodiments, the microglia is a mouse microglia. In embodiments, the microglia is a BV2 cell. In embodiments, the microglia is a human microglia. In embodiments, the microglia is a HMC3 cell. In embodiments, the glial cell is an ependymal cell.

[0098] In embodiments, the glial cell is derived from a subject to be treated with the composition (autologous). In embodiments, the glial cell is isolated from a subject to be treated with the composition. In embodiments, the glial cell is allogeneic to a subject to be treated with the composition. Methods of isolating glial cells (e.g., microglia) are well known in the art (See Wang et al., JNeurosci Methods, 2025, 423, 110558, pp. 1-7; Ryan et al., Sci Transl Med, 2017,Atty Docket No. 055523-507001WD9, 421, eaai7635, pp. 1-12; Martirosian et al., Curr Protoc, 2021, 1, 6, el40, pp. 1-13; and Rustenhoven et al., Sci Rep, 2016, 6, 19371, pp. 1-11; each of which is incorporated herein by reference in its entirety and for all purposes).

[0099] In embodiments, the glial cell is genetically modified. In some embodiments, the glial cell is genetically modified to target a target cell. In some embodiments, the target cell is a cancer cell. In some embodiments, the cancer cell is a neuroblastoma cell.

[0100] In an aspect, provided herein, is a composition containing an oncolytic virus and glial cell (e.g., microglia), wherein the glial cell is infected by the oncolytic virus.

[0101] In embodiments, the oncolytic virus is a poxvirus. In embodiments, the oncolytic virus is vaccinia virus. In embodiments, the oncolytic vaccinia virus is selected from Dryvax, ACAM1000. ACAM2000. Lister, EM63, LIVP. Tian Tan, Copenhagen, Western Reserve. Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I and Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve. Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen 1 or Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax. In embodiments, the oncolytic vaccinia virus is ACAM1000. In embodiments, the oncolytic vaccinia virus is ACAM2000. In embodiments, the oncolytic vaccinia virus is Lister. In embodiments, the oncolytic vaccinia virus is EM63. In embodiments, the oncolytic vaccinia virus is LIVP. In embodiments, the oncolytic vaccinia virus is Tian Tan. In embodiments, the oncolytic vaccinia virus is Copenhagen, Western Reserve. In embodiments, the oncolytic vaccinia virus is Modified Vaccinia Ankara (MV A). In embodiments, the oncolytic vaccinia virus is New York City Board of Health. In embodiments, the oncolytic vaccinia virus is Dairen. In embodiments, the oncolytic vaccinia virus is Ikeda. In embodiments, the oncolytic vaccinia virus is LC16M8. In embodiments, the oncolytic vaccinia vims is Tashkent. In embodiments, the oncolytic vaccinia virus is Wyeth. In embodiments, the oncolytic vaccinia vims is IHD-J. In embodiments, the oncolytic vaccinia virus is IHD-W. In embodiments, the oncolytic vaccinia virus is Brighton. In embodiments, the oncolytic vaccinia vims is Dairen I. In embodiments, the oncolytic vaccinia virus is Connaught strains.

[0102] In embodiments, the vaccinia vims does not include the Lister strain vaccinia virus. In embodiments, the vaccinia vims does not include the Lister vaccinia virus. In embodiments, the vaccinia virus does not include the LIVP 1.1.1 vaccinia vims.Atty Docket No. 055523-507001WG

[0103] In embodiments, the oncolytic virus does not lyse the glial cell. In embodiments, the virus does not lyse the glial cell for at least 1 day to at least 10 days (or any sub value or subrange therein including endpoints) after infection. In embodiments, the virus does not lyse the glial cell for at least 1 day to at least 5 days after infection. In embodiments, the vims does not lyse the glial cell for at least 1 day after infection. In embodiments, the vims does not lyse the glial cell for at least 2 day after infection. In embodiments, the virus does not lyse the glial cell for at least 3 day after infection. In embodiments, the vims does not lyse the glial cell for at least 4 day after infection. In embodiments, the vims does not lyse the glial cell for at least 5 day after infection. In embodiments, the vims does not lyse the glial cell for at least 6 day after infection. In embodiments, the virus does not lyse the glial cell for at least 7 day after infection. In embodiments, the virus does not lyse the glial cell for at least 8 day after infection. In embodiments, the virus does not lyse the glial cell for at least 9 day after infection. In embodiments, the virus does not lyse the glial cell for at least 10 day after infection. The length of time may be any value or subrange within the recited ranges.

[0104] In embodiments, the oncolytic vims (e.g., vaccinia virus) does not induce apoptosis in the glial cell (e.g., microglia). In embodiments, the oncolytic vims (e.g., vaccinia vims) does not induce necrosis in the glial cell (e.g., microglia).

[0105] In embodiments, the oncolytic vims (e.g., vaccinia virus) is capable of replicating in the glial cell (e.g., microglia). In embodiments, the oncolytic vims (e g., vaccinia vims) is capable of replicating in the glial cell (e.g., microglia) without lysing the glial cell. In embodiments, the replication of the oncolytic vims (e.g., vaccinia virus) in the glial cell is greater than the replication in anon-glial cell (e.g., cancer cell). In embodiments, the replication of the oncolytic virus (e.g., vaccinia virus) in the glial cell is greater than the replication in a cancer cell.

[0106] In embodiments, the glial cell (e.g., microglia) is capable of secreting the oncolytic virus (e.g., vaccinia virus). In embodiments, the glial cell (e.g., microglia) is capable of secreting the oncolytic virus (e.g., vaccinia vims) without being lysed. In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the glial cell (e.g., microglia) is greater than the amount secreted from a non-glial cell (e.g., cancer cell). In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the glial cell (e.g., microglia) is greater than the amount secreted from a cancer cell. In embodiments, the glial cell (e g., microglia) is capable of releasing the oncolytic vims (e.g., vaccinia virus). In embodiments, the glial cell (e.g., microglia) is capable of releasing the oncolytic virus (e.g., vaccinia virus) without being lysed. In embodiments, the amount of the oncolytic vims (e.g., vaccinia virus) released from the glial cellAtty Docket No. 055523-507001WG(e. g. , microglia) is greater than the amount released from a non-glial cell (e.g., cancer cell). In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) released from the glial cell (e.g., microglia) is greater than the amount released from a cancer cell. In embodiments, the glial cell (e.g., microglia) is capable of being a carrier of the oncolytic virus (e.g., vaccinia virus).

[0107] In embodiments, the glial cell (e.g., microglia) infected with the oncolytic virus (e.g.. vaccinia virus) retains phagocytic activity. In embodiments, the glial cell (e.g., microglia) infected with the oncolytic virus (e.g., vaccinia virus) retains cytokine secretion capacity. In embodiments, the glial cell (e.g., microglia) infected with the oncolytic virus (e.g., vaccinia virus) retains migratory behavior toward tumor cells. In embodiments, the glial cell (e.g., microglia) infected with the oncolytic virus (e.g., vaccinia virus) retains tumor-homing properties. In embodiments, the glial cell (e.g., microglia) infected with the oncolytic virus (e g., vaccinia virus) retains viability for at least 24 hours, 48 hours, 72 hours, 96 hours, 5 days, 6 days, or 7 days post-infection. In embodiments, the oncolytic virus (e.g., vaccinia virus) remains replication-competent within the glial cell (e.g., microglia) for at least 24 hours, 48 hours, 72 hours, 96 hours, 5 days, 6 days, or 7 days post-infection. In embodiments, the infected glial cell (e.g., microglia) is capable of transferring the oncolytic virus (e.g., vaccinia vims) to a cancer cell without undergoing lysis. In embodiments, the infected glial cell (e.g., microglia) is capable of releasing extracellular vesicles containing the oncolytic virus (e.g., vaccinia virus). In embodiments, the infected glial cell (e.g., microglia) is capable of forming tunneling nanotubes to transfer the oncolytic virus (e.g., vaccinia vims) to a cancer cell. In embodiments, the infected glial cell (e.g., microglia) retains metabolic activity post-infection, as measured by ATP production, mitochondrial function, or viability assays. In embodiments, the infected glial cell (e.g., microglia) retains responsiveness to inflammatory stimuli (e.g., LPS, IFNy) post-infection. In embodiments, the infected glial cell (e.g., microglia) retains the ability to interact with other immune cells (e.g., T cells, NK cells) post-infection. In embodiments, the viability of the infected glial cell (e.g., microglia) is greater than the viability of a non-glial cell (e.g., cancer cell) infected with the same oncolytic virus. In embodiments, the functional activity of the infected glial cell (e.g., microglia), including phagocytosis, cytokine secretion, or tumor-homing behavior, is greater than that of a non-glial cell (e g., cancer cell) infected with the same oncolytic virus. In embodiments, the duration of viral replication within the infected glial cell (e.g., microglia) is greater than the duration of viral replication within a non-glial cell (e.g., cancer cell). In embodiments, the amount of virus released or secreted by the infected glial cell (e.g.. microglia) is greater than the amount released or secreted by a non-glial cell (e.g., cancer cell).Atty Docket No. 055523-507001WDIV. MACROPHAGE COMPOSITIONS

[0108] Any method for making (e.g., deriving, differentiating, isolating, culturing, expanding, etc.) and using macrophages cells (e.g., CNS-associated macrophages) may be used with the compositions and methods described herein. Methods of making and using macrophage cells are well known in the art.

[0109] In an aspect, provided herein, is a composition containing an oncolytic virus and a macrophage, wherein the macrophage is infected by the oncolytic virus.

[0110] In embodiments, the macrophage is derived from a subject to be treated with the composition (autologous). In embodiments, the macrophage is isolated from a subject to be treated with the composition. In embodiments, the macrophage is allogeneic to a subject to be treated with the composition. Methods of isolating macrophages (e.g., CNS-associated macrophages) are well known in the art (See Wang et al., J Neurosci Methods, 2025, 423, 110558, pp. 1-7; Ryan et al., Sci Transl Med, 2017, 9, 421, eaai7635, pp. 1-12; and Rustenhoven et al.. Sci Rep, 2016, 6. 19371, pp. 1-11; each of which is incorporated herein by reference in its entirety and for all purposes).

[0111] In embodiments, the macrophage is genetically modified. In some embodiments, the macrophage is genetically modified to target a target cell. In some embodiments, the target cell is a cancer cell. In some embodiments, the cancer cell is a neuroblastoma cell.

[0112] In an aspect, provided herein, is a composition containing an oncolytic virus and macrophage, wherein the macrophage is infected by the oncolytic virus.

[0113] In embodiments, the oncolytic virus is a poxvirus. In embodiments, the oncolytic virus is vaccinia virus. In some embodiments, the oncolytic vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New' York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton. Dairen I and Connaught strains.

[0114] In embodiments, the vaccinia virus does not include the Lister strain vaccinia virus. In embodiments, the vaccinia virus does not include the Lister vaccinia virus. In embodiments, the vaccinia virus does not include the LIVP 1.1.1 vaccinia virus.

[0115] In embodiments, the oncolytic virus does not lyse the macrophage. In embodiments, the virus does not lyse the macrophage for at least 1 day to at least 10 days (or any sub value or subrange therein including endpoints) after infection. In embodiments, the virus does not lyse theAtty Docket No. 055523-507001WG macrophage for at least 1 day to at least 5 days after infection. In embodiments, the virus does not lyse the macrophage for at least 1 day after infection. In embodiments, the virus does not lyse the macrophage for at least 2 day after infection. In embodiments, the virus does not lyse the macrophage for at least 3 day after infection. In embodiments, the virus does not lyse the macrophage for at least 4 day after infection. In embodiments, the virus does not lyse the macrophage for at least 5 day after infection. In embodiments, the virus does not lyse the macrophage for at least 6 day after infection. In embodiments, the virus does not lyse the macrophage for at least 7 day after infection. In embodiments, the virus does not lyse the macrophage for at least 8 day after infection. In embodiments, the virus does not lyse the macrophage for at least 9 day after infection. In embodiments, the virus does not lyse the macrophage for at least 10 day after infection. The length of time may be any value or subrange within the recited ranges.

[0116] In embodiments, the oncolytic virus (e.g., vaccinia virus) does not induce apoptosis in the macrophage (e.g., CNS-associated macrophage). In embodiments, the oncolytic vims (e.g.. vaccinia virus) does not induce necrosis in the macrophage (e.g., CNS-associated macrophage).

[0117] In embodiments, the macrophage cell (e.g., CNS-associated macrophage) infected with the oncolytic virus (e.g., vaccinia vims) remains viable

[0118] In embodiments, the oncolytic vims (e.g., vaccinia virus) is capable of replicating in the macrophage (e.g., CNS-associated macrophage). In embodiments, the oncolytic vims (e.g., vaccinia virus) is capable of replicating in the macrophage (e.g., CNS-associated macrophage) without lysing the macrophage cell. In embodiments, the replication of the oncolytic virus (e.g., vaccinia virus) in the macrophage cell is greater than the replication in a non-macrophage cell (e.g., cancer cell). In embodiments, the replication of the oncolytic virus (e.g., vaccinia vims) in the macrophage cell is greater than the replication in a cancer cell.

[0119] In embodiments, the macrophage (e.g., CNS-associated macrophage) is capable of secreting the oncolytic vims (e.g., vaccinia virus). In embodiments, the macrophage (e.g., CNS- associated macrophage) is capable of secreting the oncolytic virus (e.g., vaccinia virus) without being lysed. In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the macrophage (e.g., CNS-associated macrophage) is greater than the amount secreted from a non-macrophage cell (e.g., cancer cell). In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the macrophage (e g., CNS-associated macrophage) is greater than the amount secreted from a cancer cell. In embodiments, the macrophage (e.g., CNS- associated macrophage) is capable of releasing the oncolytic virus (e.g., vaccinia virus). InAtty Docket No. 055523-507001WG embodiments, the macrophage (e.g., CNS-associated macrophage) is capable of releasing the oncolytic virus (e.g., vaccinia virus) without being lysed. In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) released from the macrophage (e.g., CNS-associated macrophage) is greater than the amount released from a non-macrophage cell (e.g., cancer cell). In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) released from the macrophage (e g., CNS-associated macrophage) is greater than the amount released from a cancer cell.

[0120] In embodiments, the macrophage (e.g., CNS-associated macrophage) infected with the oncolytic virus (e.g., vaccinia virus) retains phagocytic activity. In embodiments, the macrophage (e.g., CNS-associated macrophage) infected with the oncolytic virus (e.g., vaccinia virus) retains cytokine secretion capacity. In embodiments, the macrophage (e g., CNS-associated macrophage) infected with the oncolytic virus (e.g., vaccinia virus) retains migratory behavior toward tumor cells. In embodiments, the macrophage (e.g., CNS-associated macrophage) infected with the oncolytic virus (e.g., vaccinia virus) retains tumor-homing properties. In embodiments, the macrophage (e.g., CNS-associated macrophage) infected with the oncolytic virus (e.g., vaccinia virus) retains viability for at least 24 hours, 48 hours, 72 hours, 96 hours, 5 days, 6 days, or 7 days post-infection. In embodiments, the oncolytic virus (e.g., vaccinia virus) remains replication-competent within the macrophage (e.g., CNS-associated macrophage) for at least 24 hours, 48 hours, 72 hours, 96 hours, 5 days, 6 days, or 7 days post-infection. In embodiments, the infected macrophage (e g., CNS-associated macrophage) is capable of transferring the oncolytic virus (e.g., vaccinia virus) to a cancer cell without undergoing lysis. In embodiments, the infected macrophage (e.g., CNS-associated macrophage) is capable of releasing extracellular vesicles containing the oncolytic virus (e.g., vaccinia virus). In embodiments, the infected macrophage (e.g., CNS-associated macrophage) is capable of forming tunneling nanotubes to transfer the oncolytic virus (e.g., vaccinia virus) to a cancer cell. In embodiments, the infected macrophage (e.g., CNS-associated macrophage) retains metabolic activity post-infection, as measured by ATP production, mitochondrial function, or viability assays. In embodiments, the infected macrophage (e.g., CNS-associated macrophage) retains responsiveness to inflammatory stimuli (e.g., LPS, IFNy) post-infection. In embodiments, the infected macrophage (e g., CNS-associated macrophage) retains the ability to interact with other immune cells (e.g., T cells, NK cells) postinfection. In embodiments, the viability of the infected macrophage (e.g., CNS-associated macrophage) is greater than the viability of a non-macrophage cell (e.g., cancer cell) infected with the same oncolytic virus. In embodiments, the functional activity of the infected macrophage (e.g., CNS-associated macrophage), including phagocytosis, cytokine secretion, orAtty Docket No. 055523-507001WG tumor-homing behavior, is greater than that of a non-macrophage cell (e.g., cancer cell) infected with the same oncolytic virus. In embodiments, the duration of viral replication within the infected macrophage (e.g., CNS-associated macrophage) is greater than the duration of viral replication within a non-macrophage cell (e.g., cancer cell). In embodiments, the amount of virus released or secreted by the infected macrophage (e.g.. CNS-associated macrophage) is greater than the amount released or secreted by a non-macrophage cell (e.g., cancer cell).V. METHODS OF USETreatment of Cancer

[0121] The compositions provided herein including embodiments thereof are, inter alia, useful for the treatment of cancer. In embodiments, the methods provided herein are, inter alia, useful for treating cancer including administering glial cells (e.g., microglia) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e.g., a poxvirus or vaccinia virus). In embodiments, the methods may be used to treat glioblastoma, neuroblastoma, or other solid tumors affecting the central nervous system (CNS). In some embodiments, the oncolytic virus -infected glial cells (e.g., vaccinia virus-infected microglia) unexpectantly and efficiently delivered the virus to tumor cells and induced potent cytotoxic effects. In embodiments, the oncolytic virus-infected glial cells were effective in three-dimensional tumor models which recapitulate the in vivo tumor microenvironment. In embodiments, the methods provided herein enable targeted viral deliver}' via tumor-homing immune cells, overcoming limitations associated with poor viral penetration and immune evasion. Notably, in contrast to prior reports indicating that glial tumors in mice were not colonized by vaccinia virus due to competition between glial cells and astrocytes for viral uptake, the inventors discovered that infecting glial cells and astrocytes separately and co-culturing them with neuroblastoma cells enabled efficient viral transfer and tumor cell killing. In embodiments, the combination of the oncolytic virus and the glial cells results in a synergistic anti-cancer effect greater than either the oncolytic virus or the glial cell alone. In embodiments, the methods may be used alone or in combination with other therapies, including chemotherapy, radiotherapy, or immune checkpoint inhibitors, to enhance anti-tumor efficacy and durability of response. Thus, in an aspect is provided a method for treating cancer in a subject in need thereof, the method including administering to the subject a composition provided herein including embodiments thereof.

[0122] In another aspect is provided a use of a composition provided herein including embodiments thereof in a method of treating cancer in a subject in need thereof, the methodAtty Docket No. 055523-507001WG including administering to the subject the composition provided herein including embodiments thereof.

[0123] In another aspect is provided a use of a composition provided herein including embodiments thereof in the preparation of a medicament for treating cancer in a subj ect in need thereof, the method including administering to the subject the composition provided herein including embodiments thereof.

[0124] In embodiments, glial cells are administered in combination with the oncolytic virus. In some embodiments, the oncolytic virus infects the glial cells prior to administration. In some embodiments, the oncolytic virus infected glial cells are administered. Methods for administering glial cells are well known in the art, and can be determined by a skilled clinician.

[0125] In some embodiments, the subject is human. In embodiments, the subject is an adult patient. In embodiments, the subject is an adolescent patient. In some embodiments, the subject is a pediatric patient. In some embodiments, the subject is a neonate. In some embodiments, the subject is an infant. In some embodiments, the subject is a child. In some embodiments, the subject is an adolescent. In some embodiments, the subject is greater than 12 months in age. In some embodiments the subject is less than 18 years in age. In some embodiments the subject is greater than 18 years in age.

[0126] In embodiments, the method further includes administering to the subject a known treatment for cancer (e.g., neuroblastoma), such as a chemotherapeutic, therapeutic antibody, cancer vaccine, and the like. In embodiments, the oncolytic virus includes a recombinant polynucleotide, wherein the recombinant polynucleotide encodes a therapeutic molecule. In embodiments, the therapeutic molecule treats the cancer.

[0127] Treatments for neuroblastoma are known in the art. For example, chemotherapy, antibody treatment, bispecific antibody treatment, cancer vaccines, radiation, natural killer (NK) cells, and the like may be used. In embodiments, the method includes administering to the subject a chemotherapy to treat the neuroblastoma. In embodiments, the method includes administering one or more of: cyclophosphamide, cisplatin, carboplatin, vincristine, doxorubicin (adriamycin), etoposide, topotecan, melphalan, busulfan, or thiotepa. In embodiments, the method includes administering eflomithine (Iwilfin). In embodiments, the method includes administering 3F8 antibody. In embodiments, the method includes administering naxitamab (Hu3F8). In embodiments, the method includes administering omburtamab. In embodiments, the method includes administering HuOKT3. In embodiments, the method includes administering a vaccineAtty Docket No. 055523-507001WG targeting GD2L and GD3L. In embodiments, the method includes administering NK cells. In embodiments, the method includes administering radiation or liquid radiation therapy.

[0128] U.S. Patent No. 10.105,436, which is incorporated herein by reference in its entirety and for all purposes, describes methods of administering, making, storing, and using compositions including poxvirus and cells that can be used in the methods and compositions described herein.

[0129] In embodiments, the oncolytic virus is administered in a therapeutically effective amount, e.g., at an amount sufficient to treat the disease. In embodiments, the glial cell is administered in a therapeutically effective amount, e.g., at an amount sufficient to treat the disease. In embodiments, the oncolytic virus and glial cell are in the same (a single) composition.

[0130] In embodiments, the composition is administered to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra-parenchymal, intraventricular, intraarticular, or intraocular injection. In embodiments, the composition is administered to the subject by intravenous injection. In embodiments, the composition is administered to the subject by intraperitoneal injection. In embodiments, the composition is administered to the subject by intrathecal injection. In embodiments, the composition is administered to the subject by intra-cerebro-ventricular injection. In embodiments, the composition is administered to the subject by intrapleural injection. In embodiments, the composition is administered to the subject by intra-parenchymal injection. In embodiments, the composition is administered to the subject by intraventricular injection. In embodiments, the composition is administered to the subject by intraarticular injection. In embodiments, the composition is administered to the subject by intraocular injection.

[0131] In embodiments, the composition is administered directly to a region affected by the disease. In embodiments, the composition is administered systemically. In embodiments, the composition is administered by MRI-guided delivery.

[0132] In embodiments, the disease is cancer. In embodiments, the cancer is glioblastoma, glioma, astrocytoma, oligodendroglioma, ependymoma, medulloblastoma, central neurocytoma, primary CNS lymphoma, neuroblastoma, embryonal tumor with multilayered rosettes (ETMR), atypical teratoid / rhabdoid tumor (AT / RT), a spinal cord tumor, or a meningeal cancer. In embodiments, the cancer is glioblastoma. In embodiments, the cancer is glioma. In embodiments, the cancer is astrocytoma. In embodiments, the cancer is oligodendroglioma. In embodiments, the cancer is ependymoma. In embodiments, the cancer is medulloblastoma. In embodiments, the cancer is central neurocytoma. In embodiments, the cancer is primary CNS lymphoma. In embodiments, the cancer is neuroblastoma. In embodiments, the cancer is embryonal tumor withAtty Docket No. 055523-507001WD multilayered rosetes (ETMR). In embodiments, the cancer is atypical teratoid / rhabdoid tumor (AT / RT). In embodiments, the cancer is a spinal cord tumor. In embodiments, the cancer is a meningeal cancer.

[0133] In embodiments, the cancer is a metastatic tumor to the central nervous system. In embodiments, the cancer is a breast cancer brain metastasis, a lung cancer brain metastasis, a melanoma brain metastasis, a renal cell carcinoma brain metastasis, or a colorectal cancer brain metastasis. In embodiments, the cancer is a breast cancer brain metastasis. In embodiments, the cancer is a lung cancer brain metastasis. In embodiments, the cancer is a melanoma brain metastasis. In embodiments, the cancer is a renal cell carcinoma brain metastasis. In embodiments, the cancer is a colorectal cancer brain metastasis.

[0134] In embodiments, the oncolytic virus is a poxvirus. In embodiments, the oncolytic virus is vaccinia virus. In embodiments, the oncolytic vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton. Dairen I and Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax, AC AMI 000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I or Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax. In embodiments, the oncolytic vaccinia virus is ACAM1000. In embodiments, the oncolytic vaccinia virus is ACAM2000. In embodiments, the oncolytic vaccinia virus is Lister. In embodiments, the oncolytic vaccinia virus is EM63. In embodiments, the oncolytic vaccinia virus is LIVP. In embodiments, the oncolytic vaccinia virus is Tian Tan. In embodiments, the oncolytic vaccinia virus is Copenhagen. Western Reserve. In embodiments, the oncolytic vaccinia virus is Modified Vaccinia Ankara (MVA). In embodiments, the oncolytic vaccinia virus is New York City Board of Health. In embodiments, the oncolytic vaccinia virus is Dairen. In embodiments, the oncolytic vaccinia virus is Ikeda. In embodiments, the oncolytic vaccinia virus is LC16M8. In embodiments, the oncolytic vaccinia virus is Tashkent. In embodiments, the oncolytic vaccinia virus is Wyeth. In embodiments, the oncolytic vaccinia virus is IHD-J. In embodiments, the oncolytic vaccinia virus is IHD-W. In embodiments, the oncolytic vaccinia virus is Brighton. In embodiments, the oncolytic vaccinia virus is Dairen I. In embodiments, the oncolytic vaccinia virus is Connaught strains.Atty Docket No. 055523-507001WD

[0135] In some embodiments, vaccinia virus strain Dryvax is oncolytic. In some embodiments, vaccinia virus strain Dryvax is ACAM1000. In some embodiments, vaccinia virus strain ACAM2000 is oncolytic. In some embodiments, vaccinia virus strain Lister is oncolytic. In some embodiments, vaccinia virus strain EM63 is oncolytic. In some embodiments, vaccinia virus strain LIVP is oncolytic. In some embodiments, vaccinia virus strain Tian Tan is oncolytic. In some embodiments, vaccinia virus strain Copenhagen is oncolytic. In some embodiments, vaccinia virus strain Western Reserve is oncolytic. In some embodiments, vaccinia virus strain Modified Vaccinia Ankara (MV A) is oncolytic. In some embodiments, vaccinia virus strain New York City Board of Health is oncolytic. In some embodiments, vaccinia virus strain Dairen is oncolytic. In some embodiments, vaccinia virus strain Ikeda is oncolytic. In some embodiments, vaccinia virus strain LC16M8 is oncolytic. In some embodiments, vaccinia virus strain Tashkent is oncolytic. In some embodiments, vaccinia virus strain Wyeth is oncolytic. In some embodiments, vaccinia virus strain IHD-J is oncolytic. In some embodiments, vaccinia virus strain IHD-W is oncolytic. In some embodiments, vaccinia virus strain Brighton is oncolytic. In some embodiments, vaccinia virus strain Dairen I is oncolytic. In some embodiments, vaccinia virus strain Connaught is oncolytic.

[0136] In embodiments, the vaccinia virus does not include the Lister strain vaccinia virus. In embodiments, the vaccinia virus does not include Lister vaccinia virus. In embodiments, the vaccinia virus does not include LIVP vaccinia virus. In embodiments, the vaccinia virus does not include LIVP 1.1.1 vaccinia virus.

[0137] In embodiments, the glial cell (e.g., microglia) is infected with the oncolytic virus prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 1 day and about 10 weeks (or any sub value or subrange therein including endpoints) prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 2 days and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 3 days and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 4 days and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 5 days and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 6 days and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 1 week and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 2 weeksAtty Docket No. 055523-507001WG and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 3 weeks and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic vims between about 4 weeks and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic vims between about 5 weeks and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic virus between about 6 weeks and about 10 weeks prior to the administration to the subj ect. In embodiments, the glial cell is infected with the oncolytic vims between about 7 weeks and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic vims between about 8 weeks and about 10 weeks prior to the administration to the subject. In embodiments, the glial cell is infected with the oncolytic vims between about 9 weeks and about 10 weeks prior to the administration to the subject.

[0138] In embodiments the glial cell is infected with the oncoly tic vims at least 1 day prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 2 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic vims at least 3 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic vims at least 4 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 5 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 6 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic vims at least 7 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic vims at least 8 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 9 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 10 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic vims at least 11 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 12 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 13 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic vims at least 14 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 15 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 16 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic vims at least 17 days prior to the administration to the subject. InAtty Docket No. 055523-507001WG embodiments the glial cell is infected with the oncolytic virus at least 18 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 19 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 20 days prior to the administration to the subject. In embodiments the glial cell is infected with the oncolytic virus at least 21 days prior to the administration to the subject.

[0139] The amount of poxvirus administered to an average-sized adult can be, for example, 1 x 102to 1 x IO10plaque-forming units, 1 x 103to 1 x 108plaque-forming units, 1 x 104to 1 x 106plaque-forming units, or any value or sub range there between. As a specific example, about 2.5 x 105plaque-forming units can be used.

[0140] In embodiments, the amount of poxvirus administered is between about 1 x IO2and about 1 x IO10plaque-forming units (or any sub value or subrange therein including endpoints). In embodiments, the amount of poxvirus administered is between about 1.5 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2.5 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 102and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 5 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 102and about l x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 102and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 102and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 102and about 1 x IO10plaque-forming units. In embodiments.Atty Docket No. 055523-507001WG the amount of poxvirus administered is between about 9.5 x 102and about 1 x IO10plaqueforming units.

[0141] In embodiments, the amount of poxvirus administered is between about 1 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 1.5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2.5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x 103and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 103and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 6 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x103and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9.5 x 103and about 1 x IO10plaque-forming units.

[0142] In embodiments, the amount of poxvirus administered is between about 1 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 1.5 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2.5 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x104and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 104and about 1 x IO10plaque-forming units. InAtty Docket No. 055523-507001WG embodiments, the amount of poxvirus administered is between about 4 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 104and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 6 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 104and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x104and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 104and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9.5 x 104and about 1 x IO10plaque-forming units.

[0143] In embodiments, the amount of poxvirus administered is between about 1 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 1.5 x 105and about l x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 103and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2.5 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x105and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 105and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 6 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 105and about 1 x 1010Atty Docket No. 055523-507001WG plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x105and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 105and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9.5 x 105and about 1 x IO10plaque-forming units.

[0144] In embodiments, the amount of poxvirus administered is between about 1 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 1.5 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2.5 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x106and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 106and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 6 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 106and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x 106and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 106and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9.5 x 106and about 1 x IO10plaque-forming units.

[0145] In embodiments, the amount of poxvirus administered is between about 1 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 1.5 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 107and about 1 x IO10plaque-forming units. InAtty Docket No. 055523-507001WG embodiments, the amount of poxvirus administered is between about 2.5 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x 107and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 107and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 6 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 107and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x107and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 107and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9.5 x 107and about 1 x IO10plaque-forming units.

[0146] In embodiments, the amount of poxvirus administered is between about 1 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 1.5 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2.5 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x108and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 108and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 6 x 108Atty Docket No. 055523-507001WG and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 108and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x108and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 108and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9.5 x 108and about 1 x IO10plaque-forming units.

[0147] In embodiments, the amount of poxvirus administered is between about 1 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 1.5 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 2.5 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3 x109and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 3.5 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 4.5 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 5.5 x 109and about 1 x IO10plaqueforming units. In embodiments, the amount of poxvirus administered is between about 6 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 6.5 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 7.5 x 109and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8 x 109and about 1 x 1010plaque-forming units. In embodiments, the amount of poxvirus administered is between about 8.5 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9 x 109and about 1 x IO10plaque-forming units. In embodiments, the amount of poxvirus administered is between about 9.5 x 109and about 1 x IO10plaque-forming units.Atty Docket No. 055523-507001WG

[0148] In embodiments, the glial cells (e.g., microglia) are administered in an amount between about 1 x 104and about 1 x IO10cells (or any sub value or subrange therein including endpoints). In embodiments, the glial cells are administered in an amount between about 1.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8.5 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9 x 104and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9.5 x 104and about 1 x IO10cells.

[0149] In embodiments, the glial cells are administered in an amount between about 1 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 1.5 x 103and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2.5 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3.5 x 103and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4.5 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5 x 103and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5.5 x 105and about 1 x IO10cells. In embodiments, theAtty Docket No. 055523-507001WG glial cells are administered in an amount between about 6 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6.5 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7.5 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8.5 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9 x 105and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9.5 x 105and about 1 x IO10cells.

[0150] In embodiments, the glial cells are administered in an amount between about 1 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 1.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8 x IO6and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8.5 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9 x 106and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9.5 x 106and about 1 x IO10cells.

[0151] In embodiments, the glial cells are administered in an amount between about 1 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount betweenAtty Docket No. 055523-507001WG about 1.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8.5 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9 x 107and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9.5 x 107and about 1 x IO10cells.

[0152] In embodiments, the glial cells are administered in an amount between about 1 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 1.5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2.5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3.5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4.5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5.5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6.5 x 108and about 1Atty Docket No. 055523-507001WO x IO10cells. In embodiments, the glial cells are administered in an amount between about 7 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7.5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8.5 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9 x 108and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9.5 x 108and about 1 x IO10cells.

[0153] In embodiments, the glial cells are administered in an amount between about 1 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 1.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 2.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 3.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 4.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 5.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 6.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 7.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 8.5 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9 x 109and about 1 x IO10cells. In embodiments, the glial cells are administered in an amount between about 9.5 x 109and about 1 x IO10cells.

[0154] PCT Patent Publication No. WO 2020 / 247385 is hereby incorporated by reference in its entirety for all that is taught therein.Atty Docket No. 055523-507001WG

[0155] In embodiments, the oncolytic virus (e.g., a poxvirus or a vaccinia virus) and the glial cell (e.g., microglia) and / or macrophage (e.g., CNS-associated macrophage) are administered in a combined synergistic amount, wherein the combined synergistic amount is effective to treat cancer in a subj ect in need thereof.

[0156] In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4. 0.5, 0.6, 0.7, 0.8. 0.9, 1.0, 1.1, 1.2. 1.3, 1.4, 1.5. 1.6. 1.7, 1.8, 1.9, 2.0. 2.1, 2.2, 2.3. 2.4. 2.5, 2.6, 2.7. 2.8. 2.9, 3.0,3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2,5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4,7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6,9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17. 18. 19. 20. 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the oncolytic virus when used separately from the glial cell and / or macrophage. In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7,1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9,4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1,6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3,8.4, 8.5, 8.6, 8.7. 8.8, 8.9, 9.0, 9.1. 9.2, 9.3, 9.4, 9.5. 9.6, 9.7, 9.8, 9.9. 10.0. 11. 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, or 99% of the amount of the glial cell and / or macrophage when used separately from the oncolytic virus.

[0157] In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0,3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2,5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4. 6.5, 6.6, 6.7, 6.8. 6.9, 7.0, 7.1, 7.2. 7.3, 7.4,7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6,9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69. 70. 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84. 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95. 96. 97. 98. or 99% of the amount of the oncolytic virus when used separately from the microglia. In embodiments, a synergistic amount may beAtty Docket No. 055523-507001WG about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3,4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5,6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7,8.8, 8.9, 9.0, 9.1. 9.2, 9.3, 9.4, 9.5. 9.6, 9.7, 9.8, 9.9. 10.0. 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72,73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the microglia when used separately from the oncolytic virus.

[0158] In embodiments, the synergistic amount is about 0.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g., microglia). In embodiments, the synergistic amount is about 0.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.7% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.1% of the amount of the oncolytic vims when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 1.2% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.3% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.4% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.6% of the amount of the oncolytic virus when used separately from the glialAtty Docket No. 055523-507001WG cell (e.g. microglia). In embodiments, the synergistic amount is about 1.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.8% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 1.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0159] In embodiments, the synergistic amount is about 2. 1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.2% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 2.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.4% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 2.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, theAtty Docket No. 055523-507001WG synergistic amount is about 3.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0160] In embodiments, the synergistic amount is about 4.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.9% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 5.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.9% of theAtty Docket No. 055523-507001WG amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0161] In embodiments, the synergistic amount is about 6. 1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.6% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 7.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). InAtty Docket No. 055523-507001WG embodiments, the synergistic amount is about 8.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0162] In embodiments, the synergistic amount is about 8.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.4% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 8.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.8% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 9.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 10.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).Atty Docket No. 055523-507001WG

[0163] In embodiments, the synergistic amount is about 11% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 12% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 13% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 14% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 15% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 16% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 17% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 18% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 19% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 20% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 21% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 22% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 23% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 24% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 25% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 26% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 27% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 28% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 29% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 30% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0164] In embodiments, the synergistic amount is about 31% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergisticAtty Docket No. 055523-507001WG amount is about 32% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 33% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 34% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 35% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 36% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 37% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 38% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 39% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 40% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 41% of the amount of the oncolytic virus w hen used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 42% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 43% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 44% of the amount of the oncolytic virus wdien used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 45% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 46% of the amount of the oncolytic virus w hen used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 47% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 48% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 49% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 50% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0165] In embodiments, the synergistic amount is about 51% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 52% of the amount of the oncolytic virus w hen used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 53% of the amount of theAtty Docket No. 055523-507001WG oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 54% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 55% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 56% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 57% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 58% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 59% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 60% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 61% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 62% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 63% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 64% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 65% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 66% of the amount of the oncolytic virus w hen used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 67% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 68% of the amount of the oncolytic virus w hen used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 69% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 70% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0166] In embodiments, the synergistic amount is about 71 % of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 72% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 73% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 74% of the amount of the oncolytic virus w hen used separately fromAtty Docket No. 055523-507001WG the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 75% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 76% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 77% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 78% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 79% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 80% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 81% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 82% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 83% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 84% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 85% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 86% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 87% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 88% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 89% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 90% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 91% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 92% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 93% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 94% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 95% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 96% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). InAtty Docket No. 055523-507001WG embodiments, the synergistic amount is about 97% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 98% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 99% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0167] In embodiments, the synergistic effect may be a cancer-treating effect such as a glioblastoma (i.e., a glioblastoma-treating synergistic effect), astrocytoma (i.e., an astrocytomatreating synergistic effect), oligodendroglioma (i.e., an oligodendroglioma-treating synergistic effect), ependymoma (i.e., an ependymoma-treating synergistic effect), medulloblastoma (i.e., a medulloblastoma-treating synergistic effect), central neurocytoma (i.e.. a central neurocytomatreating synergistic effect), primary CNS lymphoma (i.e., a primary CNS lymphoma-treating synergistic effect), neuroblastoma (i.e., a neuroblastoma-treating synergistic effect), embry onal tumor with multilayered rosettes (ETMR) (i.e., an ETMR-treating synergistic effect), atypical teratoid / rhabdoid tumor (AT / RT) (i.e.. an AT / RT-treating synergistic effect), spinal cord tumor (i.e., a spinal cord tumor-treating synergistic effect), or meningeal cancer (i.e., a meningeal cancer-treating synergistic effect). In embodiments, the synergistic effect may be a cancertreating effect such as a glioblastoma (i.e., a glioblastoma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as an astrocytoma (i.e., an astrocytoma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as an oligodendroglioma (i.e., an oligodendroglioma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as an ependymoma (i.e., an ependymoma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a medulloblastoma (i.e., a medulloblastoma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a central neurocytoma (i.e., a central neurocytoma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a primary CNS lymphoma (i.e., a primary CNS lymphoma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a neuroblastoma (i.e., a neuroblastoma-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as an embryonal tumor with multilayered rosettes (ETMR) (i.e., an ETMR-treating synergistic effect). In embodiments, the synergistic effect may be a cancertreating effect such as an atypical teratoid / rhabdoid tumor (AT / RT) (i.e., an AT / RT-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a spinal cord tumor (i.e., a spinal cord tumor-treating synergistic effect). In embodiments, theAtty Docket No. 055523-507001WG synergistic effect may be a cancer-treating effect such as a meningeal cancer (i.e., a meningeal cancer-treating synergistic effect).

[0168] In embodiments, the synergistic effect may be a metastatic CNS tumor-treating effect such as a breast cancer brain metastasis (i.e., a breast cancer brain metastasis-treating synergistic effect), lung cancer brain metastasis (i.e., a lung cancer brain metastasis-treating synergistic effect), melanoma brain metastasis (i.e., a melanoma brain metastasis-treating synergistic effect), renal cell carcinoma brain metastasis (i.e., a renal cell carcinoma brain metastasis-treating synergistic effect), or colorectal cancer brain metastasis (i.e., a colorectal cancer brain metastasistreating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a breast cancer brain metastasis (i.e., a breast cancer brain metastasis-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a lung cancer brain metastasis (i.e., a lung cancer brain metastasis-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a melanoma brain metastasis (i.e., a melanoma brain metastasis-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a renal cell carcinoma brain metastasis (i.e., a renal cell carcinoma brain metastasis-treating synergistic effect). In embodiments, the synergistic effect may be a cancer-treating effect such as a colorectal cancer brain metastasis (i.e., a colorectal cancer brain metastasis-treating synergistic effect).

[0169] In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0. 1% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 0.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g.,Atty Docket No. 055523-507001WG microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 0.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 1.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 1.1% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 1.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 1.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 1.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 1.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g.Atty Docket No. 055523-507001WG microglia) results in about 1.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 1.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 1.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 1.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0170] In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.1% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.6%Atty Docket No. 055523-507001WG greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 2.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 3.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 3.1% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 3.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 3.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually7and separately . In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 3.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 3.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 3.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g.. microglia) when usedAtty Docket No. 055523-507001WG individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 3.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 3.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 3.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0171] In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4. 1% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e g. microglia) results in about 4.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. InAtty Docket No. 055523-507001WG embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 4.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e g. microglia) results in about 5.1% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.7% greater anti-cancer effect than theAtty Docket No. 055523-507001WG sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 5.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0172] In embodiments, sy nergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.1% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately . In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.7% greater anti-cancer effect than the sum of the anti-cancer effectAtty Docket No. 055523-507001WG of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 6.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.1% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e g. microglia) results in about 7.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between theAtty Docket No. 055523-507001WG oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 7.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0173] In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.1% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy’ between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 8.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vacciniaAtty Docket No. 055523-507001WG virus) and the glial cell (e.g. microglia) results in about 8.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 8.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9. 1% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9.2% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) yvhen used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 9.3% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9.4% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9.5% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 9.6% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately . In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9.7% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9.8% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic vims (e.g.,Atty Docket No. 055523-507001WG vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 9.9% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e g. microglia) results in about 10.0% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0174] In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 11% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 12% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 13% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 14% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic vims (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 15% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 16% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 17% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 18% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually andAtty Docket No. 055523-507001WG separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 19% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 20% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 21% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 22% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic vims (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 23% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 24% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 25% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 26% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic vims (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 27% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 28% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 29% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vacciniaAtty Docket No. 055523-507001WG virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 30% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0175] In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 31% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 32% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 33% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 34% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 35% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 36% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 37% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 38% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 39% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) whenAtty Docket No. 055523-507001WG used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 40% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 41% greater anti-cancer effect than the sum of the anti -cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 42% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 43% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 44% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 45% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 46% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 47% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 48% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 49% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 50% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolyticAtty Docket No. 055523-507001WG virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0176] In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 51% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic vims (e.g.. vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 52% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 53% greater anti-cancer effect than the sum of the anti -cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 54% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic vims (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 55% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 56% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 57% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 58% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic vims (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 59% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic vims (e.g.. vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 60% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cellAtty Docket No. 055523-507001WG(e.g. , microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 61% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 62% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 63% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 64% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g.. microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 65% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 66% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 67% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 68% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 69% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy’ between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 70% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.Atty Docket No. 055523-507001WG

[0177] In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 71% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 72% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 73% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g.. vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 74% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic vims (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 75% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 76% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 77% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 78% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic vims (e.g. vaccinia vims) and the glial cell (e.g. microglia) results in about 79% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic vims (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 80% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia vims) or the glial cell (e g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 81% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vacciniaAtty Docket No. 055523-507001WG virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 82% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 83% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 84% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 85% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 86% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 87% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 88% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 89% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy' between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 90% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually7and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 91% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e g., vaccinia virus) or the glial cell (e.g., microglia) when used individually7and separately. In embodiments, synergy7between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 92% greater anti-cancer effectAtty Docket No. 055523-507001WG than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 93% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 94% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 95% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 96% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 97% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 98% greater anti-cancer effect than the sum of the anti-cancer effect of the oncolytic virus (e.g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately. In embodiments, synergy between the oncolytic virus (e.g. vaccinia virus) and the glial cell (e.g. microglia) results in about 99% greater anti-cancer effect than the sum of the anticancer effect of the oncolytic virus (e g., vaccinia virus) or the glial cell (e.g., microglia) when used individually and separately.

[0178] In embodiments, the oncolytic virus (e.g., vaccinia virus) does not induce apoptosis in the glial cell (e g., microglia). In embodiments, the oncolytic virus (e.g., vaccinia virus) does not induce necrosis in the glial cell (e.g., microglia).

[0179] In embodiments, the oncolytic virus (e.g., vaccinia virus) is capable of replicating in the glial cell (e.g., microglia). In embodiments, the oncolytic virus (e.g., vaccinia virus) is capable of replicating in the glial cell (e.g., microglia) without lysing the glial cell. In embodiments, the replication of the oncolytic virus (e.g., vaccinia virus) in the glial cell is greater than the replication in anon-glial cell (e.g., cancer cell). In embodiments, the replication of the oncolytic virus (e.g., vaccinia virus) in the glial cell is greater than the replication in a cancer cell.Atty Docket No. 055523-507001WG

[0180] In embodiments, the glial cell (e.g., microglia) is capable of secreting the oncolytic virus (e.g., vaccinia virus). In embodiments, the glial cell (e.g., microglia) is capable of secreting the oncolytic virus (e.g., vaccinia vims) without being lysed. In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the glial cell (e.g., microglia) is greater than the amount secreted from a non-glial cell (e.g., cancer cell). In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the glial cell (e.g., microglia) is greater than the amount secreted from a cancer cell. In embodiments, the glial cell (e.g., microglia) is capable of releasing the oncolytic vims (e.g., vaccinia virus). In embodiments, the glial cell (e.g., microglia) is capable of releasing the oncolytic virus (e.g., vaccinia virus) without being lysed. In embodiments, the amount of the oncolytic vims (e.g., vaccinia virus) released from the glial cell (e.g., microglia) is greater than the amount released from a non-glial cell (e.g., cancer cell). In embodiments, the amount of the oncolytic vims (e.g., vaccinia virus) released from the glial cell (e.g., microglia) is greater than the amount released from a cancer cell.VI. COMPOSITIONS

[0181] The compositions provided herein, including embodiments thereof, generally relate to the compositions including glial cells (e.g., microglia) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e.g., a poxvirus or vaccinia virus). In embodiments, the compositions may be. inter alia, formulated for use in treating cancer, including glioblastoma, neuroblastoma, or other solid tumors affecting the central nervous system (CNS). In embodiments, the compositions include oncolytic virus-infected glial cells capable of delivering the vims to tumor cells and inducing potent cytotoxic effects. In embodiments, the compositions are effective in three-dimensional tumor models that recapitulate the in vivo tumor microenvironment. Notably, in contrast to prior reports indicating that glial tumors were not colonized by vaccinia virus due to competition between glial cells and astrocytes for viral uptake, the inventors discovered that infecting these cell types separately enabled efficient viral transmission and tumor cell killing. In embodiments, the combination of the oncolytic virus and the glial cells results in a synergistic anti-cancer effect greater than either the oncolytic virus or the glial cell alone. In embodiments, the compositions may be used alone or in combination with other therapeutic agents, including chemotherapeutics, radiotherapeutics, or immune checkpoint inhibitors. In embodiments, the compositions may be formulated for direct administration or for ex vivo delivery’ using autologous or allogeneic immune cells. Thus, in an aspect is provided a composition including an oncolytic virus and a glial cell, wherein the glial cell is infected by the oncolytic virusAtty Docket No. 055523-507001WD

[0182] In one aspect, provided herein is a composition including an oncolytic virus (e.g. a poxvirus, e.g., smallpox, for example a smallpox vaccine) and a glial cell. In another aspect, provided herein is a composition including an oncolytic virus (e.g., a poxvirus, e.g., smallpox, for example a small pox vaccine) and a macrophage (e.g., CNS-associated macrophage). In embodiments, the oncolytic virus is a poxvirus. In embodiments, the oncolytic virus is a vaccinia virus. In embodiments, the oncolytic virus comprises a recombinant polynucleotide, wherein said recombinant polynucleotide encodes a therapeutic molecule.

[0183] In embodiments, the glial cell is an oligodendrocyte, astrocyte, microglia, or ependymal cell. In embodiments, the glial cell is an oligodendrocyte. In embodiments, the glial cell is an astrocyte. In embodiments, the glial cell is a microglia. In embodiments, the microglia is a mouse microglia. In embodiments, the microglia is a BV2 cell. In embodiments, the microglia is a human microglia. In embodiments, the microglia is aHMC3 cell. In embodiments, the glial cell is an ependymal cell.

[0184] In embodiments, the oncolytic virus is a poxvirus. In embodiments, the oncolytic virus is vaccinia virus. In embodiments, the oncolytic vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I and Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I or Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax. In embodiments, the oncolytic vaccinia virus is ACAM1000. In embodiments, the oncolytic vaccinia vims is ACAM2000. In embodiments, the oncolytic vaccinia virus is Lister. In embodiments, the oncolytic vaccinia virus is EM63. In embodiments, the oncolytic vaccinia vims is LIVP. In embodiments, the oncolytic vaccinia vims is Tian Tan. In embodiments, the oncolytic vaccinia virus is Copenhagen, Western Reserve. In embodiments, the oncolytic vaccinia vims is Modified Vaccinia Ankara (MV A). In embodiments, the oncolytic vaccinia vims is New York City Board of Health. In embodiments, the oncolytic vaccinia vims is Dairen. In embodiments, the oncolytic vaccinia virus is Ikeda. In embodiments, the oncolytic vaccinia vims is LC16M8. In embodiments, the oncolytic vaccinia vims is Tashkent. In embodiments, the oncolytic vaccinia virus is Wyeth. In embodiments, the oncolytic vaccinia virus is IHD-J. In embodiments, the oncolytic vaccinia virus is IHD-W. In embodiments, the oncolytic vaccinia virus is Brighton. InAtty Docket No. 055523-507001WG embodiments, the oncolytic vaccinia virus is Dairen I. In embodiments, the oncolytic vaccinia virus is Connaught strains.

[0185] In some embodiments, vaccinia virus strain Dryvax is oncolytic. In some embodiments, vaccinia virus strain Dryvax is ACAM1000. In some embodiments, vaccinia virus strain ACAM2000 is oncolytic. In some embodiments, vaccinia virus strain Lister is oncolytic. In some embodiments, vaccinia virus strain EM63 is oncolytic. In some embodiments, vaccinia virus strain LIVP is oncolytic. In some embodiments, vaccinia virus strain Tian Tan is oncolytic. In some embodiments, vaccinia virus strain Copenhagen is oncolytic. In some embodiments, vaccinia virus strain Western Reserve is oncolytic. In some embodiments, vaccinia virus strain Modified Vaccinia Ankara (MV A) is oncolytic. In some embodiments, vaccinia virus strain New York City Board of Health is oncolytic. In some embodiments, vaccinia virus strain Dairen is oncolytic. In some embodiments, vaccinia virus strain Ikeda is oncolytic. In some embodiments, vaccinia virus strain LC16M8 is oncolytic. In some embodiments, vaccinia virus strain Tashkent is oncolytic. In some embodiments, vaccinia virus strain Wyeth is oncolytic. In some embodiments, vaccinia virus strain IHD-J is oncolytic. In some embodiments, vaccinia virus strain IHD-W is oncolytic. In some embodiments, vaccinia virus strain Brighton is oncolytic. In some embodiments, vaccinia virus strain Dairen I is oncolytic. In some embodiments, vaccinia virus strain Connaught is oncolytic.

[0186] In embodiments, the vaccinia virus does not include the Lister strain vaccinia virus. In embodiments, the vaccinia virus does not include Lister vaccinia virus. In embodiments, the vaccinia virus does not include LIVP vaccinia virus. In embodiments, the vaccinia virus does not include LIVP 1.1.1 vaccinia virus.

[0187] In embodiments, the oncolytic virus (e.g., a poxvirus or a vaccinia virus) and the glial cell (e.g.. microglia) and / or macrophage (e.g., CNS-associated macrophage) are present in a combined synergistic amount, wherein the combined synergistic amount is effective to treat a disease (e g., cancer) in a subject in need thereof. In embodiments, the combined synergistic amount is a combined therapeutically effective amount for treating cancer in a subject in need thereof. In embodiments, the combined synergistic amount includes an amount of the oncolytic virus (e.g., poxvirus or vaccinia virus) that is less than a therapeutically effective amount of the oncolytic virus when administered alone as a single agent. In embodiments, the combined synergistic amount includes an amount of a glial cell (e.g. microglia) and / or macrophage (e.g., CNS-associated macrophage) that is less than a therapeutically effective amount of the glial cell and / or macrophage when administered alone as a single agent. In embodiments, the combinedAtty Docket No. 055523-507001WD synergistic amount includes an amount of oncolytic virus and an amount of a glial cell and / or macrophage that are each less than a therapeutically effective amount of the oncolytic virus and the therapeutically effective amount of the glial cell and / or macrophage when each of the compounds (e.g.. oncolytic virus and the glial cell and / or macrophage) are administered alone as a single agent. In embodiments, the therapeutically effective amount of the oncolytic virus (e.g., poxvirus or vaccinia virus) is an U.S. Food and Drug Administration (FDA)-approved amount of oncolytic virus that is effective in treating a disease, disorder, or condition in a subject in need thereof. In embodiments, the therapeutically effective amount of a glial cell and / or macrophage is an FDA-approved amount of a glial cell and / or macrophage that is effective in treating a disease, disorder, or condition in a subject in need thereof. In embodiments, the combined therapeutically effective amount is a therapeutically effective amount of oncolytic virus combined with a therapeutically effective amount of a glial cell and / or macrophage. In embodiments, the synergistic effect may be an anti-cancer effect.

[0188] In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4. 0.5, 0.6, 0.7, 0.8. 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0,3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2,5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4,7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6,9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17. 18. 19. 20. 21. 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the oncolytic virus when used separately from the glial cell and / or macrophage. In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7,1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9,4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1,6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7. 7.8, 7.9, 8.0, 8.1. 8.2, 8.3,8.4, 8.5, 8.6, 8.7. 8.8, 8.9, 9.0, 9.1. 9.2, 9.3, 9.4, 9.5. 9.6, 9.7, 9.8, 9.9. 10.0. 11. 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80. 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, or 99% of the amount of the glial cell and / or macrophage when used separately from the oncolytic virus.Atty Docket No. 055523-507001WG

[0189] In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0,3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2,5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4,7.5, 7.6, 7.7, 7.8. 7.9, 8.0, 8.1, 8.2. 8.3, 8.4, 8.5, 8.6. 8.7, 8.8, 8.9, 9.0. 9.1, 9.2, 9.3, 9.4. 9.5, 9.6,9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95. 96. 97. 98, or 99% of the amount of the oncolytic virus when used separately from the microglia. In embodiments, a synergistic amount may be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1,2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3,4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9. 6.0, 6.1, 6.2, 6.3. 6.4, 6.5,6.6, 6.7, 6.8, 6.9. 7.0, 7.1, 7.2, 7.3. 7.4, 7.5, 7.6, 7.7. 7.8, 7.9, 8.0, 8.1. 8.2, 8.3, 8.4, 8.5. 8.6, 8.7,8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58. 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72,73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84. 85. 86. 87. 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amount of the microglia when used separately from the oncolytic virus.

[0190] In embodiments, the synergistic amount is about 0.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g., microglia). In embodiments, the synergistic amount is about 0.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.4% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.5% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.7% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 0.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.0% of the amount of theAtty Docket No. 055523-507001WG oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 1.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0191] In embodiments, the synergistic amount is about 2. 1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.4% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.5% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.7% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 2.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.1% of the amount of the oncolytic vims when used separately fromAtty Docket No. 055523-507001WG the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 3.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0192] In embodiments, the synergistic amount is about 4.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.4% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 4.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 4.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5. 1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). InAtty Docket No. 055523-507001WG embodiments, the synergistic amount is about 5.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 5.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0193] In embodiments, the synergistic amount is about 6. 1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 6.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.1% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 7.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount isAtty Docket No. 055523-507001WG about 7.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.5% of the amount of the oncolytic virus w en used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 7.8% of the amount of the oncolytic vims when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 7.9% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0194] In embodiments, the synergistic amount is about 8. 1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.4% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 8.5% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 8.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.0% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.1% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.2% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.3% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.4% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.5% of the amount of the oncolyticAtty Docket No. 055523-507001WG virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.6% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.7% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.8% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 9.9% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 10.0% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia).

[0195] In embodiments, the synergistic amount is about 11% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 12% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 13% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 14% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 15% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 16% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 17% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 18% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 19% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 20% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 21% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 22% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 23% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 24% of the amount of the oncolytic vims when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 25% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 26% of the amount of the oncolytic vims when used separately from the glialAtty Docket No. 055523-507001WG cell (e.g. microglia). In embodiments, the synergistic amount is about 27% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 28% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 29% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 30% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0196] In embodiments, the synergistic amount is about 31% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 32% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 33% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 34% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 35% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 36% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 37% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 38% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 39% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 40% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 41% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 42% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 43% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 44% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 45% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 46% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 47% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, theAtty Docket No. 055523-507001WG synergistic amount is about 48% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 49% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 50% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0197] In embodiments, the synergistic amount is about 51% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 52% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 53% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 54% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 55% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 56% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 57% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 58% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 59% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 60% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 61% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 62% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 63% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 64% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 65% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 66% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 67% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 68% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 69% of theAtty Docket No. 055523-507001WG amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 70% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0198] In embodiments, the sy nergistic amount is about 71% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 72% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 73% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 74% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 75% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 76% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 77% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 78% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 79% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 80% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 81% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 82% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 83% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 84% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 85% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 86% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 87% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 88% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 89% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 90% of the amount of the oncolytic virus whenAtty Docket No. 055523-507001WG used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 91% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 92% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 93% of the amount of the oncolytic virus when used separately from the glial cell (e g. microglia). In embodiments, the synergistic amount is about 94% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 95% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 96% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 97% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 98% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia). In embodiments, the synergistic amount is about 99% of the amount of the oncolytic virus when used separately from the glial cell (e.g. microglia).

[0199] In embodiments, the oncolytic virus (e.g., vaccinia virus) does not induce apoptosis in the glial cell (e.g., microglia). In embodiments, the oncolytic virus (e.g., vaccinia virus) does not induce necrosis in the glial cell (e.g., microglia).

[0200] In embodiments, the oncolytic virus (e.g., vaccinia virus) is capable of replicating in the glial cell (e.g., microglia). In embodiments, the oncolytic virus (e.g., vaccinia virus) is capable of replicating in the glial cell (e.g., microglia) without lysing the glial cell. In embodiments, the replication of the oncolytic vims (e.g., vaccinia virus) in the glial cell is greater than the replication in anon-glial cell (e.g., cancer cell). In embodiments, the replication of the oncolytic virus (e.g., vaccinia virus) in the glial cell is greater than the replication in a cancer cell.

[0201] In embodiments, the glial cell (e.g., microglia) is capable of secreting the oncolytic virus (e.g., vaccinia virus). In embodiments, the glial cell (e.g., microglia) is capable of secreting the oncolytic virus (e.g., vaccinia vims) without being lysed. In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the glial cell (e.g., microglia) is greater than the amount secreted from a non-glial cell (e.g., cancer cell). In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) secreted from the glial cell (e.g., microglia) is greater than the amount secreted from a cancer cell. In embodiments, the glial cell (e.g., microglia) is capable of releasing the oncolytic vims (e.g., vaccinia virus). In embodiments, the glial cell (e.g., microglia) is capable of releasing the oncolytic virus (e.g., vaccinia virus) without being lysed. InAtty Docket No. 055523-507001WG embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) released from the glial cell (e.g., microglia) is greater than the amount released from a non-glial cell (e.g., cancer cell). In embodiments, the amount of the oncolytic virus (e.g., vaccinia virus) released from the glial cell (e.g., microglia) is greater than the amount released from a cancer cell.

[0202] In embodiments, the composition is formulated for administration to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra- parenchymal, intraventricular, intraarticular, or intraocular injection. In embodiments, the composition is formulated for administration to the subject by intravenous injection. In embodiments, the composition is formulated for administration to the subject by intraperitoneal injection. In embodiments, the composition is formulated for administration to the subject by intrathecal injection. In embodiments, the composition is formulated for administration to the subject by intra-cerebro-ventricular injection. In embodiments, the composition is formulated for administration to the subject by intrapleural injection. In embodiments, the composition is formulated for administration to the subject by intra-parenchymal injection. In embodiments, the composition is formulated for administration to the subject by intraventricular injection. In embodiments, the composition is formulated for administration to the subject by intraarticular injection. In embodiments, the composition is formulated for administration to the subject by intraocular injection.

[0203] In some embodiments, the compositions disclosed herein comprise a pharmaceutically acceptable carrier. As used herein, the term “pharmaceutically acceptable carrier” refers to solvents, diluents, preservatives, dispersion or suspension aids, isotonic agents, thickening or emulsifying agents, solid binders, and lubricants, appropriate for the particular dosage form. The skilled artisan is aware of a variety of different carriers that may be used in formulating pharmaceutical compositions and knows techniques for the preparation thereof (See Remington’s Pharmaceutical Sciences Ed. by Gennaro, Mack Publishing, Easton, Pa., 1995; which is incorporated herein in its entirety by reference). The pharmaceutically acceptable carriers may include, but are not limited to Ringer’s solution, isotonic saline, starches, potato starch, sugars, glucose, powdered tragacant, malt, gelatin, talc, cellulose and its derivatives, ethyl cellulose, sodium carboxymethyl cellulose, cellulose acetate excipients, cocoa butter, suppository waxes, agar, alginic acid, oils, cottonseed oil, peanut oil, safflower oil, sesame oil, olive oil, soybean oil, com oil, glycols, propylene glycol, esters, ethyl laureate, ethyl oleate, buffering agents, aluminum hydroxide, magnesium hydroxide, phosphate buffer solutions, pyrogen-free water, ethyl alcohol, other non-toxic compatible lubricants, sodium lauryl sulfate, magnesium stearate, coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents.Atty Docket No. 055523-507001WGPharmaceutically acceptable carriers may also include preservatives and antioxidants. One or more of the above-mentioned materials can be specifically excluded from the compositions and methods of some embodiments.

[0204] The effective dosage of each of the treatment modalities disclosed herein may vary depending on various factors, including but not limited to the particular treatment, compound or pharmaceutical composition employed, the mode of administration, the condition being treated, and / or the severity of the condition being treated. Thus, the dosage regimen of the combination of the invention is selected in accordance wi th a variety7of factors including the route of administration and the renal and hepatic function of the patient. A physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the single active ingredients required to prevent, counter or arrest the progress of the condition. Optimal precision in achieving concentration of the active ingredients within the range that yields efficacy without toxicity requires a regimen based on the kinetics of the active ingredients' availability to target sites.

[0205] Methods of preparing pharmaceutical compositions including the relevant treatments disclosed herein are known in the art and will be apparent from the art, from known standard references, such as Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 18th edition (1990), which is incorporated herein by reference in its entirety.

[0206] It should be understood that the embodiments described herein are not limited to vaccinations or vaccinating per se. but also relate to generating an immune response or reaction to an antigen associated with a disease. While the words “vaccine,” “vaccination,” or other like terms are used for convenience, it should be understood that such embodiments also relate to immune compositions, immunogenic compositions, immune response generation, immunization, etc., where absolute prophylactic immunity is not required or generated. For example, the embodiments referring to vaccination also can relate to generating or to assisting in creating an immunogenic or immune response against an antigen, regardless of whether that response results in absolute eradication or immunization against the disease to be treated.

[0207] U.S. Patent No. 10,105,436, which is incorporated herein by reference in its entirety7, describes methods of administering, making, storing, and using compositions including poxvirus and cells that can be used in the methods and compositions described herein.

[0208] In another aspect is provided a delivery device. In embodiments, the delivery device is configured to store and / or administer compositions comprising glial cells (e.g., microglia) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e.g., aAtty Docket No. 055523-507001WG poxvirus or vaccinia virus). In embodiments, the delivery device is configured to store and administer compositions comprising glial cells (e.g., microglia) and / or macrophages (e.g., CNS- associated macrophages) infected with an oncolytic virus (e.g., a poxvirus or vaccinia virus). In embodiments, the delivery device is configured to store or administer compositions comprising glial cells (e.g.. microglia) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e.g., a poxvirus or vaccinia virus). In embodiments, the delivery device is configured to store compositions comprising glial cells (e.g., microglia) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e g., a poxvirus or vaccinia virus). In embodiments, the delivery device is configured to administer compositions comprising glial cells (e.g., microglia) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e.g., a poxvirus or vaccinia virus).

[0209] In embodiments, the deliver}' device may be a syringe, catheter, implantable reservoir, or other biocompatible container. In embodiments, the delivery device is suitable for maintaining cell viability and / or viral infectivity prior to administration. In embodiments, the device may include temperature control features, cell suspension media, or encapsulation matrices to preserve the therapeutic integrity of the infected cells during storage and transport. In embodiments, the device may be configured for intravenous, intraperitoneal, intrathecal, intra- cerebro-ventricular, intrapleural, intra-parenchymal. intraventricular, intraarticular, or intraocular administration (e.g., injection). In embodiments, the device may include a controlled-release mechanism to enable sustained or localized deliver}' of the infected cells over time. In embodiments, the device may be pre-loaded with a defined dose of infected cells and sealed under sterile conditions for single-use or multi-dose applications. In embodiments, the device may further include components for imaging guidance, such as radiopaque markers or MRI- compatible materials, to facilitate precise delivery to CNS tumor sites.VII. METHODS OF MANUFACTURE

[0210] Provided herein are methods of making (e.g., manufacturing or producing) the compositions provided herein including embodiments thereof. The methods of making provided herein, including embodiments thereof, generally relate to the preparation of therapeutic compositions including glial cells (e g., microglia) and / or macrophages (e.g., CNS-associated macrophages) infected with an oncolytic virus (e.g., a poxvirus or vaccinia virus). In embodiments, the methods include isolating glial cells (e.g., microglia) and / or macrophages (e g., CNS-associated macrophages) from a subject. In embodiments, the methods further includeAtty Docket No. 055523-507001WO infecting the isolated glial cells and / or isolated macrophages with an oncolytic virus under conditions that preserve cell viability and viral infectivity. In embodiments, the manufacturing methods enable the generation of cell-based delivery7vehicles capable of targeted viral transfer to tumor cells. Notably, in contrast to prior reports indicating that glial tumors were not colonized by vaccinia virus due to competition between glial cells and astrocytes for viral uptake, the inventors discovered that infecting these cell types separately enabled efficient viral transmission and downstream tumor cell killing. In embodiments, the methods include culturing infected glial cells under conditions that promote viral replication and packaging. In embodiments, the manufacturing methods may further include steps for purification, formulation, and quality control to ensure therapeutic potency and reproducibility. Thus, in an aspect is provided a method for making a composition including a poxvirus and glial cells, the method including: (a) obtaining glial cells from a subject; and (b) contacting the glial cells with a poxvirus to form poxvirus-infected glial cells.

[0211] In embodiments, the glial cells are isolated prior to step b).

[0212] In embodiments, the glial cells include oligodendrocytes, astrocytes, microglia cells, ependymal cells, or a mixture thereof.

[0213] In embodiments, the glial cells include astrocytes, microglia cells, or a mixture thereof.

[0214] In another aspect is provided a method for making a composition including a poxvirus and macrophages, the method including: (a) obtaining macrophages from a subject; and (b) contacting the macrophages with a poxvirus to form poxvirus-infected macrophages. In embodiments, the macrophages are CNS-associated macrophages.

[0215] In embodiments, the poxvirus is a vaccinia virus. In embodiments, the vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City7Board of Health, Dairen, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J, and IHD-W, Brighton, Dairen I and Connaught strains. In embodiments, the vaccinia virus is a Lister strain. In embodiments, the vaccinia virus is a Copenhagen strain.

[0216] In embodiments, the glial cells are infected with the poxvirus at a titer betw een > 0.5 MOI and > 5 MOI (or any sub value or subrange therein including endpoints). In embodiments, the glial cells are infected with the poxvirus at a titer > 0.5 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 0.6 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 0.7 MOI. In embodiments, the glial cells are infected with theAtty Docket No. 055523-507001WO poxvirus at a titer > 0.8 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 0.9 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.1 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.2 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.3 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.4 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.5 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.6 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.7 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.8 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 1.9 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.1 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.2 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.3 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.4 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.5 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.6 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.7 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.8 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 2.9 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.1 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.2 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.3 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.4 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.5 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.6 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.7 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.8 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 3.9 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.1 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.2 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.3 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.4 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.5 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.6 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.7 MOI. In embodiments, the glial cells are infected with the poxvirus at aAtty Docket No. 055523-507001WG titer > 4.8 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 4.9 MOI. In embodiments, the glial cells are infected with the poxvirus at a titer > 5 MOI.

[0217] In embodiments, the glial cells are incubated with the virus for about 1 day to about 5 days (or any sub value or subrange therein including endpoints). In embodiments, the glial cells are incubated with the virus for about 2 days to about 5 days. In embodiments, the glial cells are incubated with the virus for about 3 days to about 5 days. In embodiments, the glial cells are incubated with the virus for about 4 days to about 5 days.In embodiments, the glial cells are incubated with the virus for about 1 day to about 4 days. In embodiments, the glial cells are incubated with the virus for about 1 day to about 3 days. In embodiments, the glial cells are incubated with the virus for about 1 day to about 2 days.

[0218] In embodiments, the glial cells are incubated with the virus for about 1 day. In embodiments, the glial cells are incubated with the virus for about 2 days. In embodiments, the glial cells are incubated with the virus for about 3 days. In embodiments, the glial cells are incubated with the virus for about 4 days. In embodiments, the glial cells are incubated with the virus for about 5 days. In embodiments, the glial cells are incubated with the virus for about 6 days. In embodiments, the glial cells are incubated with the virus for about 7 days. In embodiments, the glial cells are incubated with the virus for about 8 days. In embodiments, the glial cells are incubated with the virus for about 9 days. In embodiments, the glial cells are incubated with the virus for about 10 days. In embodiments, the glial cells are incubated with the virus for about 11 days. In embodiments, the glial cells are incubated with the virus for about 12 days. In embodiments, the glial cells are incubated with the virus for about 13 days. In embodiments, the glial cells are incubated with the virus for about 14 days. In embodiments, the glial cells are incubated with the virus for about 15 days. In embodiments, the glial cells are incubated with the virus for about 16 days. In embodiments, the glial cells are incubated with the virus for about 17 days. In embodiments, the glial cells are incubated with the virus for about 18 days. In embodiments, the glial cells are incubated with the virus for about 19 days. In embodiments, the glial cells are incubated with the virus for about 20 days. In embodiments, the glial cells are incubated with the virus for about 21 days.

[0219] In embodiments, the poxvirus does not lyse the glial cells and / or the CNS-associated macrophages. In embodiments, the poxvirus does not lyse the glial cells and the CNS-associated macrophages. In embodiments, the poxvirus does not lyse the glial cells or the CNS-associated macrophages. In embodiments, the poxvirus does not lyse the glial cells. In embodiments, the poxvirus does not lyse the CNS-associated macrophages.Atty Docket No. 055523-507001WD

[0220] In embodiments, the oncolytic virus (e.g., vaccinia virus) does not induce apoptosis in the glial cell (e.g., microglia). In embodiments, the oncolytic virus (e.g., vaccinia virus) does not induce necrosis in the glial cell (e.g., microglia).

[0221] In embodiments, the glial cells are derived from a subject to be treated with the composition. In embodiments, the glial cells are allogeneic to a subject to be treated with the composition.

[0222] In embodiments, the method includes an in vitro co-culture model system. In embodiments, the method includes 2D (Two-Dimensional) monolayers and / or 3D (Three- Dimensional) spheroid models. In embodiments, the method includes 2D (Two-Dimensional) monolayers and 3D (Three-Dimensional) spheroid models. In embodiments, the method includes 2D (Two-Dimensional) monolayers or 3D (Three-Dimensional) spheroid models. In embodiments, the method includes 2D (Two-Dimensional) monolayers. In embodiments, the method includes 3D (Three-Dimensional) spheroid models.

[0223] In embodiments, the oncolytic virus is a poxvirus. In embodiments, the oncolytic virus is vaccinia virus. In embodiments, the oncolytic vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, L1VP. Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I and Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve. Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Tashkent, Wyeth, IHD-J, IHD-W, Brighton, Dairen I or Connaught strains. In embodiments, the oncolytic vaccinia virus is Dryvax. In embodiments, the oncolytic vaccinia virus is ACAM1000. In embodiments, the oncolytic vaccinia vims is ACAM2000. In embodiments, the oncolytic vaccinia virus is Lister. In embodiments, the oncolytic vaccinia virus is EM63. In embodiments, the oncolytic vaccinia vims is LIVP. In embodiments, the oncolytic vaccinia vims is Tian Tan. In embodiments, the oncolytic vaccinia virus is Copenhagen, Western Reserve. In embodiments, the oncolytic vaccinia vims is Modified Vaccinia Ankara (MV A). In embodiments, the oncolytic vaccinia vims is New York City Board of Health. In embodiments, the oncolytic vaccinia vims is Dairen. In embodiments, the oncolytic vaccinia virus is Ikeda. In embodiments, the oncolytic vaccinia vims is LC16M8. In embodiments, the oncolytic vaccinia vims is Tashkent. In embodiments, the oncolytic vaccinia virus is Wyeth. In embodiments, the oncolytic vaccinia vims is IHD-J. In embodiments, the oncolytic vaccinia virus is IHD-W. In embodiments, the oncolytic vaccinia virus is Brighton. InAtty Docket No. 055523-507001WG embodiments, the oncolytic vaccinia virus is Dairen I. In embodiments, the oncolytic vaccinia virus is Connaught strains.

[0224] In some embodiments, vaccinia virus strain Dryvax is oncolytic. In some embodiments, vaccinia virus strain Dryvax is ACAM1000. In some embodiments, vaccinia virus strain ACAM2000 is oncolytic. In some embodiments, vaccinia virus strain Lister is oncolytic. In some embodiments, vaccinia virus strain EM63 is oncolytic. In some embodiments, vaccinia virus strain LIVP is oncolytic. In some embodiments, vaccinia virus strain Tian Tan is oncolytic. In some embodiments, vaccinia virus strain Copenhagen is oncolytic. In some embodiments, vaccinia virus strain Western Reserve is oncolytic. In some embodiments, vaccinia virus strain Modified Vaccinia Ankara (MV A) is oncolytic. In some embodiments, vaccinia virus strain New York City Board of Health is oncolytic. In some embodiments, vaccinia virus strain Dairen is oncolytic. In some embodiments, vaccinia virus strain Ikeda is oncolytic. In some embodiments, vaccinia virus strain LC16M8 is oncolytic. In some embodiments, vaccinia virus strain Tashkent is oncolytic. In some embodiments, vaccinia virus strain Wyeth is oncolytic. In some embodiments, vaccinia virus strain IHD-J is oncolytic. In some embodiments, vaccinia virus strain IHD-W is oncolytic. In some embodiments, vaccinia virus strain Brighton is oncolytic. In some embodiments, vaccinia virus strain Dairen I is oncolytic. In some embodiments, vaccinia virus strain Connaught is oncolytic.

[0225] In embodiments, the vaccinia virus does not include the Lister strain vaccinia virus. In embodiments, the vaccinia virus does not include Lister vaccinia virus. In embodiments, the vaccinia virus does not include LIVP vaccinia virus. In embodiments, the vaccinia virus does not include LIVP 1.1.1 vaccinia virus.

[0226] In embodiments, the composition is formulated for administration to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra- parenchymal, intraventricular, intraarticular, or intraocular injection. In embodiments, the composition is formulated for administration to the subject by intravenous injection. In embodiments, the composition is formulated for administration to the subject by intraperitoneal injection. In embodiments, the composition is formulated for administration to the subject by intrathecal injection. In embodiments, the composition is formulated for administration to the subject by intra-cerebro-ventricular injection. In embodiments, the composition is formulated for administration to the subject by intrapleural injection. In embodiments, the composition is formulated for administration to the subject by intra-parenchymal injection. In embodiments, the composition is formulated for administration to the subject by intraventricular injection. InAtty Docket No. 055523-507001WG embodiments, the composition is formulated for administration to the subject by intraarticular injection. In embodiments, the composition is formulated for administration to the subject by intraocular injection.

[0227] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety7for all purposes.EXAMPLESExample 1: Preparation of Glial cells

[0228] Microglia, the resident macrophages of the central nervous system (CNS), serve as the brain's first line of defence by engulfing harmful pathogens, including viruses, and clearing cellular debris, misfolded proteins, and dying cells to maintain homeostasis. Vaccinia virus (VV), a member of the poxvirus family, is recognized for its strong cancer killing and immunomodulatory properties. Neuroblastoma is the most prevalent cancer in infants and the third most common cancer in children after leukaemia and brain tumours. Previous studies reported that glial tumors in mice were not colonized by Vaccinia Virus due to the competition of the glial cells and astrocytes for tumor infecting viruses. To overcome this inability7to kill brain tumors by Vaccinia Virus, glial cells and astrocyte cells were infected separately and added to neuroblastoma cells using an in vitro co-culture system.

[0229] Method: In order to ensure significant infection without causing undue cytotoxicity, mouse microglial cells (BV2) were infected with VV at the optimum multiplicity7of infection (MOI). Both infected and uninfected microglia were then co-cultured separately with human neuroblastoma cell lines (SK-N-AS and SH-SY5Y). Cell viability7, proliferation, and apoptosis of neuroblastoma were assessed using MTT assays and the InCucyte visualization system. Additionally, virus release from BV2 cells was quantified using the plaque assay method. A 3D co-culture model was also developed using the hanging drop method.

[0230] Results: VV-infected microglia were capable of releasing live VV. Co-culture of these infected microglia wi th neuroblastoma cells resulted in a significant reduction of viability and proliferation compared to uninfected controls. Furthermore, the 3D co-culture model with VV- infected microglia showed a drastic decrease in the number of tumour cells. All results were included.Atty Docket No. 055523-507001WO

[0231] Conclusion: VV infection of microglia triggers a response that significantly impairs the viability and survival of neuroblastoma cells due to virus replication and oncolysis. This study underscores the potential of utilizing virus-infected microglia as career of oncolytic vaccinia to alter the tumor microenvironment and suppress tumour cell growth by oncolysis.Example 2; Vaccinia virus converts microglia into potent oncolytic agent for glioblastoma and neuroblastoma

[0232] Abstract

[0233] Microglia, the resident macrophages of the central nervous system (CNS), constitute the brain’s first line of defense. Vaccinia virus (VV), a member of the poxvirus family, is well known for its potent oncolytic and immunomodulatory properties, and a favorable safety profile from historical smallpox vaccination campaign. Neuroblastoma is the most common cancer in infants and the third most frequent pediatric malignancy after leukemia and brain tumors. Glioblastoma (GBM), the most aggressive primary brain tumor in adults, carries a dismal prognosis. In previous in vivo studies, it was observed that glial tumors were not effectively colonized by VV, likely due to competition from resident glial cells and astrocytes for viral uptake. To address this limitation, an in vitro co-culture model system was developed, using both 2D (Two-Dimensional) monolayers and 3D (Three-Dimensional) spheroid models, in which VV- infected microglia cells were targeted to neuroblastoma and glioblastoma cells, enabling direct viral delivery to tumor targets. This study found that microglia BV2 cells supported productive VV replication, efficiently transferred infectious virus to tumor cells, and accelerated virolysis in both 2D and 3D systems. In spheroid cultures, microglia carriers promoted deeper viral penetration and more rapid structural collapse compared with free viruses. Therefore, pre-loading microglia with VV overcomes previously observed glial cell-mediated sequestration of virus, enabling potent delivery to brain tumor cells.

[0234] Background

[0235] Oncolytic viruses (OVs) are a class of viruses capable of selectively attacking, infecting, and destroying malignant cells within the tumor microenvironment, thereby slowing or halting tumor progression [1], These viruses, whether naturally occurring or genetically engineered, replicate preferentially in cancer cells, sparing healthy tissues, and exert direct cytotoxic effects. Unlike gene therapy, which uses viral vectors solely as delivery vehicles for therapeutic genes, OV therapy harnesses the virus itself as the active therapeutic agent [2], Several OVs have been developed from diverse viral backbones, including vaccinia virus, herpes simplex virus, and adenovirus, each with unique properties and therapeutic advantages [3],Atty Docket No. 055523-507001WG

[0236] VV, a member of the Poxviridae family, has atracted particular interest in oncolytic virotherapy owing to its well-established safety record - hundreds of millions of individuals were immunized with various VV strains during the Smallpox Eradication Program (1950-1980) [4], VV demonstrates a unique capacity to selectively infect and lyse cancer cells while largely sparing normal cells. It replicates rapidly, initiating robust viral production within six hours postinfection, and can infect a wide spectrum of malignancies, likely due to its large repertoire of viral proteins mediating cell entry [5, 6], Depending on the tumor cell type, most infected cells undergo lysis within 48-72 hours.

[0237] An important feature of VV biology is its cytoplasmic replication cycle, which prevents integration of the viral genome into host DNA. During replication, VV produces a small proportion of extracellular virus (EV) particles, infectious virions with a distinct structure incorporating host-derived membrane proteins, that facilitate evasion of the complement system and enable spread to distant metastases [7-9], These features make VV one of the most extensively studied viruses to date.

[0238] Challenges in VV therapy remain, particularly in delivering the virus efficiently to tumor sites. Intravenously administered VV must navigate multiple host immune barriers, including complement proteins, coagulation factors, blood cell components, and pre-existing neutralizing antibodies (nAbs), to target effectively the tumor mass [10, 11],

[0239] To enhance therapeutic efficacy, VV is often combined with other anticancer treatments or delivered via cellular carriers, which can both circumvent host immune defenses and boost antitumor immunity

[0012] ,

[0240] Neuroblastoma is the most common malignancy of infancy and the third most frequent pediatric cancer, responsible for approximately 1 % of childhood cancer-related deaths. Arising from postganglionic sympathetic nervous system cells, most often in the adrenal gland, neuroblastoma is diagnosed before age 10 in 96% of cases [13, 14], Current treatment regimens ty pically span 18 months and may include chemotherapy, surgical resection, high-dose chemotherapy with autologous stem cell rescue, radiotherapy, immunotherapy, and isotretinoin.

[0241] GBM, the most common malignant brain tumor in adults, is a highly aggressive astrocytic tumor with a median survival of only 12-15 months despite multimodal therapy

[0015] , Approximately 13.000 Americans are diagnosed annually. Stand...

Claims

Atty Docket No. 055523-507001WGWHAT IS CLAIMED IS:

1. A composition comprising an oncolytic virus and a glial cell, wherein the glial cell is infected by the oncolytic virus.

2. The composition of claim 1, wherein the glial cell is an oligodendrocyte, astrocyte, microglia cell, or ependymal cell.

3. The composition of claim 2, wherein the glial cell is a microglia cell, or an astrocyte.

4. The composition of any one of claims 1 to 3, wherein the glial cell is derived from a subject to be treated with the composition.

5. The composition of any one of claims 1 to 3, wherein the glial cell is allogeneic to a subject to be treated with the composition.

6. The composition of any one of claims 1 to 5, wherein the glial cell is genetically modified.

7. The composition of claim 6, wherein the glial cell is genetically modified to target a cancer cell.

8. The composition of any one of claims 1 to 7. wherein the virus does not lyse the glial cell for at least 5 days after infection.

9. The composition of any one of claims 1 to 8, comprising a plurality of glial cells infected by the oncolytic virus.

10. The composition of claim 9, wherein the plurality7of glial cells comprises oligodendrocytes, astrocytes, microglia cells, ependymal cells, or a mixture thereof.

11. The composition of any one of claims 1 to 10, wherein the oncolytic virus is a poxvirus.

12. The composition of claim 11. wherein the poxvirus is a vaccinia virus.Atty Docket No. 055523-507001WD13. The composition of claim 12, wherein the vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63. LIVP, Tian Tan, Copenhagen. Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J, and IHD-W, Brighton, Dairen I and Connaught strains.

14. The composition of claim 13, wherein the vaccinia virus is a Lister strain.

15. The composition of claim 13, wherein the vaccinia virus is a Copenhagen strain.

16. A pharmaceutical composition comprising the composition of any one of claims 1 to 15 and a pharmaceutically acceptable excipient.

17. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject a composition of any one of claims 1 to 16.

18. The method of claim 17, wherein the cancer is neuroblastoma.

19. The method of claim 17 or 18, wherein the composition is administered to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra-parenchymal, intraventricular, intraarticular, or intraocular injection.

20. The method of any one of claims 17 to 19, wherein the composition is administered directly to a region affected by the disease.

21. The method of any one of claims 17 to 20, wherein the composition is administered by MRI-guided delivery.

22. The method of any one of claims 17 to 21, wherein the subject is a human.

23. A method for making a composition comprising a poxvirus and glial cells, the method comprising:(a) obtaining glial cells from a subject; and(b) contacting the glial cells with a poxvirus to form poxvirus -infected glial cells.Atty Docket No. 055523-507001WO24. The method of claim 23, wherein the glial cells are isolated prior to step b).

25. The method of claim 23 or 24, wherein the glial cells comprise oligodendrocytes, astrocytes, microglia cells, ependymal cells, or a mixture thereof.

26. The method of claim 25, wherein the glial cells comprise astrocytes, microglia cells, or a mixture thereof.

27. The method of any one of claims 23 to 26, wherein the poxvirus is a vaccinia virus.

28. The method of claim 27, wherein the vaccinia virus is selected from Dryvax. ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MV A), New York City Board of Health, Dairen, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent. Tian Tan, Wyeth, IHD-J. and IHD-W. Brighton, Dairen I and Connaught strains.

29. The method of claim 28, wherein the vaccinia virus is a Lister strain.

30. The method of claim 28, wherein the vaccinia virus is a Copenhagen strain.

31. The method of any one of claims 23 to 30, wherein the glial cells are infected with the poxvirus at a titer > 0.5 MOI.

32. The method of claim 31, wherein the glial cells are infected with the poxvirus at a titer > 1 MOI.

33. The method of any one of claims 23 to 32, wherein the glial cells are incubated with the virus for about 1 day to about 5 days.

34. The method of claim 33, wherein the glial cells are incubated with the virus for about 3 days.Atty Docket No. 055523-507001WG35. The method of any one of claims 23 to 34, wherein the poxvirus does not lyse the glial cells.

36. The method of any one of claims 23 to 35, wherein the glial cells are derived from a subject to be treated with the composition.

37. The method of any one of claims 23 to 25, wherein the glial cells are allogeneic to a subject to be treated with the composition.

38. Use of a composition of any of claims 1 to 16 in a method of treating cancer in a subject in need thereof, the method comprising administering to the subject the composition of any one of claims 1 to 16.

39. The use of claim 38, wherein the cancer is neuroblastoma or glioma.

40. The use of claim 38 or 39, wherein the composition is administered to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra- parenchymal, intraventricular, intraarticular, or intraocular injection.

41. The use of any one of claims 38 to 40, wherein the composition is administered directly to a region affected by the disease.

42. The use of any one of claims 38 to 41, wherein the composition is administered by MRI-guided delivery.

43. The use of any one of claims 38 to 42, wherein the subject is a human.

44. Use of a composition of any of claims 1 to 16 in the preparation of a medicament for treating cancer in a subject in need thereof, the method comprising administering to the subject the composition of any one of claims 1 to 16.

45. The use of claim 44, wherein the cancer is neuroblastoma or glioma.

46. The use of claim 44 or 45, wherein the composition is administered to the subject by intravenous, intraperitoneal, intrathecal, intra-cerebro-ventricular, intrapleural, intra- parenchymal, intraventricular, intraarticular, or intraocular injection.Atty Docket No. 055523-507001WG47. The use of any one of claims 44 to 46, wherein the composition is administered directly to a region affected by the disease.

48. The use of any one of claims 44 to 47, wherein the composition is administered by MRI-guided delivery.

49. The use of any one of claims 44 to 48, wherein the subject is a human.

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