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Oncolytic Herpes Simplex病毒（OHSV）对胶质母细胞瘤（GBM）进行有效治疗几乎可以肯定 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力。这 肿瘤微环境（TME）必须通过募集来支持适应性免疫调查 活性肿瘤抗原呈现细胞和响应式CD8+ T细胞。但是，GBM肿瘤众所周知“冷” 代谢善于预防促炎性TME的肿瘤。这种免疫 - 抑制性GBM TME表现出（1）检查点蛋白质Tigit和PD-1的更新表达 相应的抑制性配体，（2）胞外酶CD39和CD73增加了局部生产 腺苷（ADO）；两种途径赋予免疫抑制和肿瘤侵略性。在这里我们建议 改善RQNESTIN34.5V.1 A f菌株OHSV-1的抗肿瘤活性。该病毒已被设计为允许 自然病毒基因的肿瘤特异性表达挫败了先天性抗病毒细胞反应（例如ICP34.5）。 RQNESTIN34.5V.2是删除GFP的RQNESTIN34.5V.1的修改版本，在患者中被证明是安全的 试验具有长时间肿瘤内病毒抗原表达的证据（NCT03152318）。三 目的旨在评估RQNESTIN34.5V.1患者衍生型（PDX）模型的增强和 同步GBM小鼠模型。我们建议在AIM 1中检验以下假设。 RQNESTIN34.5V.1将表现出增强的溶瘤活性，增强病毒传播和持久性 瘤。在AIM 2中，我们建议检验以下假设：RQNESTIN34.5V.1的治疗效率可以是 通过使用IL-12组合武装载体，检查点抑制剂（CPI）抗PD-1和抗 - 可改善 Tigit（AIM 2A）或腺苷脱氨酶（ADA）基因（AIM 2B）。 ADA武装OHSV的效率将 与表达PTEN和抗CD73（OHSV-P10-CD73; Project 3）和抗CD73的OHSV结合进行测试 根据结果​​，两个武装基因将被引入单个载体以提高治疗效率 研究。人类临床试验数据的分析使项目2能够定义一个TME曲线特征 RQNESTIN34.5V2响应者证明了患者反应与TCR多样性增加之间的相关性。 在AIM 3中，我们将检验以下假设：RQNESTIN34.5V.1和武装衍生物将增加积累 效应T细胞群体的诱导T细胞的克隆膨胀，这些细胞识别病毒和肿瘤特异性 抗原。 AIM 3分析将将AIM 2中产生的武装向量与项目3产生的武装向量进行比较 和4，并且将使我们能够相对于克隆T细胞建立多个OHSV变体的相关性 膨胀和动物生存。提出的矢量修改可能会大大改善RQNESTIN34.5V.1 性能并有效增加对OHSV治疗的肿瘤数量，从而有可能 提供适合临床开发的新型矢量。