Project 1: Treatment of GBM using an oncolytic HSV engineered to improve immunogenic tumor destruction

项目 1：使用经过改造的溶瘤 HSV 治疗 GBM，以改善免疫原性肿瘤破坏

• 批准号: 10712280
• 负责人: Joseph C Glorioso
• 金额: $ 34.32万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-07 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712280
• 关键词: Adenosine; Affect; Aggressive behavior; American; Animals; Antigen-Presenting Cells; Antitumor Response; Area; Biological Assay; CD8-Positive T-Lymphocytes; Cell Death; Cell Line; Cell Separation; Cells; Characteristics; Clinic; Clinical Trials; Clonal Expansion; Collaborations; Data; Development; Dissociation; Engineering; Evaluation; Exhibits; Genes; Glioblastoma; Glycoproteins; Goals; Growth; HSV vector; Heterogeneity; Human; Immune; Immune checkpoint inhibitor; Immunosuppression; In Vitro; Infiltration; Inflammatory; Interleukin-12; Ligands; Macrophage; Malignant Glioma; Malignant neoplasm of brain; Mediating; Metabolic; Modeling; Modification; Monitor; Mus; Mutation; Myeloid Cells; Myeloid-derived suppressor cells; Nature; Oncolytic; Oncolytic viruses; Outcome; PD-1 inhibitors; PTEN gene; PVRL1; Pathway interactions; Patients; Performance; Phenotype; Population; Production; Proteins; Receptor Cell; Recruitment Activity; Recurrence; Resistance; Safety; Simplexvirus; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Time; Transgenes; Translating; Treatment Efficacy; Tumor Antigens; Tumor Cell Line; Tumor Immunity; Variant; Vertebral column; Viral; Viral Antigens; Viral Genes; Viral Load result; Virus; Virus Replication; adaptive immune response; adenosine deaminase; anti-cancer; arm; cell killing; clinical development; combinatorial; design; effective therapy; effector T cell; efficacy evaluation; efficacy study; exhaust; immunogenic; improved; in vitro activity; in vivo; in vivo evaluation; insight; mouse model; mutant; neoplastic cell; novel; oncolysis; oncolytic herpes simplex virus; oncolytic virotherapy; patient derived xenograft model; patient response; pre-clinical; prevent; programmed cell death protein 1; recruit; response; single-cell RNA sequencing; tumor; tumor microenvironment; vector

PROJECT SUMMARY – PROJECT 1 The effective treatment of glioblastoma (GBM) using oncolytic Herpes Simplex Virus (oHSV) will almost certainly require both broad intratumoral virus spread and tumor destruction and virus-induced anti-tumor immunity. The tumor microenvironment (TME) must be supportive of adaptive immune responses through the recruitment of active tumor antigen presenting cells and responsive CD8+ T cells. However, GBM tumors are notoriously “cold” tumors that are metabolically adept at preventing the development of a pro-inflammatory TME. This immune- suppressive GBM TME exhibits upregulated expression of (1) the checkpoint proteins TIGIT and PD-1, and their respective inhibitory ligands, and (2) the ectoenzymes CD39 and CD73 that increase local production of adenosine (ADO); both pathways confer immunosuppression and tumor aggression. Here we propose to improve the anti-tumor activity of rQNestin34.5v.1 an F strain oHSV-1. This virus has been engineered to allow tumor-specific expression of natural viral genes that thwart innate anti-viral cellular responses (e.g. ICP34.5). rQNestin34.5v.2, a modified version of rQNestin34.5v.1 with GFP removed, was shown to be safe in a patient trial with evidence of intratumoral viral antigen expression for extended time periods (NCT03152318). Three aims are designed to evaluate rQNestin34.5v.1 enhancement in patient derived xenograft (PDX) models and in syngeneic GBM mouse models. We propose in Aim 1, to test the hypothesis that syncytial mutant variants of rQNestin34.5v.1 will exhibit enhanced oncolytic activity, augmenting viral spread and persistence within the tumor. In Aim 2, we propose to test the hypothesis that the therapeutic efficacy of rQNestin34.5v.1 can be improved by arming the vector with a combination of IL-12, the checkpoint inhibitors (CPI) anti-PD-1 and anti- TIGIT (Aim 2A), or with the adenosine deaminase (ADA) gene (Aim 2B). The efficacy of ADA-armed oHSV will be tested in combination with oHSV expressing PTEN and anti-CD73 (oHSV-P10-CD73; Project 3) and depending on the outcome, both arming genes will be introduced into a single vector for treatment efficacy studies. Analysis of human clinical trial data has allowed Project 2 to define a TME profile characteristic of rQNestin34.5v2 responders, demonstrating a correlation between patient response and increased TCR diversity. In Aim 3, we will test the hypothesis that rQNestin34.5v.1 and armed derivatives will increase the accumulation of effector T cell populations and induce clonal expansion of T cells that recognize viral- and tumor-specific antigens. Aim 3 analyses will compare the armed vectors generated in Aim 2 to those generated by Projects 3 and 4 and will allow us to establish correlations across multiple oHSV variants with respect to clonal T cell expansion and animal survival. The proposed vector modifications may substantially improve rQNestin34.5v.1 performance and effectively increase the number of tumors responsive to oHSV treatment, thereby potentially providing a novel vector suitable for clinical development.

项目摘要-项目1 使用溶瘤性单纯疱疹病毒（oHSV）有效治疗胶质母细胞瘤（GBM）几乎肯定 需要广泛的肿瘤内病毒扩散和肿瘤破坏以及病毒诱导的抗肿瘤免疫。的 肿瘤微环境（TME）必须通过募集 活性肿瘤抗原呈递细胞和应答性CD 8 + T细胞。然而，GBM肿瘤是出了名的“冷酷” 在代谢上擅长阻止促炎性TME发展的肿瘤。这种免疫- 抑制性GBM TME表现出（1）检查点蛋白TIGIT和PD-1的上调表达，以及它们的 各自的抑制配体，和（2）增加局部产生的胞外酶CD 39和CD 73， 腺苷（ADO）;这两种途径赋予免疫抑制和肿瘤侵袭。在此，我们建议 提高rQNestin34.5v.1和F株oHSV-1的抗肿瘤活性。这种病毒被设计成 天然病毒基因的肿瘤特异性表达，其阻碍先天性抗病毒细胞应答（例如ICP34.5）。 rQNestin34.5v.2是rQNestin34.5v.1的修改版本，删除了GFP，已被证明对患者是安全的 具有肿瘤内病毒抗原长时间表达证据的试验（NCT 03152318）。三 目的是评价rQNestin34.5v.1在患者来源的异种移植物（PDX）模型中的增强作用， 同系GBM小鼠模型。我们在目的1中提出，为了检验以下假设，即， rQNestin34.5v.1将表现出增强的溶瘤活性，增加病毒在肿瘤细胞内的扩散和持久性。 肿瘤在目的2中，我们提出检验以下假设：rQNestin34.5v.1的治疗功效可以是 通过用IL-12、检查点抑制剂（CPI）抗PD-1和抗PD-2抗体的组合武装载体来改善。 TIGIT（Aim 2A）或腺苷脱氨酶（ADA）基因（Aim 2B）。ADA武装的oHSV的疗效将 与表达PTEN和抗-CD 73的oHSV组合测试（oHSV-P10-CD 73;项目3），和 根据结果，两个武装基因将被引入到单个载体中以获得治疗功效。 问题研究对人体临床试验数据的分析使项目2能够定义以下TME特征： rQNestin34.5v2应答者，证明了患者应答与TCR多样性增加之间的相关性。 在目标3中，我们将检验rQNestin34.5v.1和武装衍生物将增加蓄积的假设 并诱导识别病毒和肿瘤特异性抗原的T细胞的克隆扩增。 抗原目标3分析将比较目标2中生成的武装矢量和项目3中生成的武装矢量 并且将允许我们建立多个oHSV变体之间关于克隆T细胞的相关性。 扩张和动物生存。所提出的载体修饰可以显著改善rQNestin34.5v.1 有效地增加对oHSV治疗有反应的肿瘤数量，从而潜在地 提供了一种适于临床开发的新型载体。