Overcoming Barriers of Virotherapy by Next-Generation oHSV Expressing E-Cadherin

通过表达 E-钙粘蛋白的下一代 oHSV 克服病毒治疗的障碍

• 批准号: 10544483
• 负责人: Jianhua Yu
• 金额: $ 37.49万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544483
• 关键词: Address; Animals; Binding; Biology; Cell Adhesion Molecules; Cell fusion; Cell physiology; Cells; Complex; Creativeness; Cytolysis; Data; Diagnosis; Dose; E-Cadherin; Eating; Engineering; FDA approved; Fc Immunoglobulins; Glioblastoma; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Herpesvirus 1; Immune; Immune response; Immunity; Immunocompetent; Immunoglobulin G; Immunoglobulins; Immunologic Deficiency Syndromes; Immunosuppression; Infection; Innate Immune Response; Killer Cells; Laboratories; Ligands; Malignant Neoplasms; Manuscripts; Mediating; Minor; Modeling; Mus; N-Cadherin; NK Cell Activation; Names; Natural Killer Cells; Neoplasm Metastasis; Normal Cell; Oncolytic viruses; Operative Surgical Procedures; Paper; Patients; Penetration; Pharmaceutical Preparations; Play; Production; Publications; Publishing; Radiation therapy; Reporting; Reproduction; Research; Role; Signal Transduction; Site; Solid Neoplasm; Surface; Testing; Toxic effect; Viral; Virotherapy; Virus; anti-tumor immune response; blood-brain barrier crossing; cancer therapy; chemotherapy; chimeric antigen receptor; cytotoxicity; immune clearance; improved; in vivo; interest; neoplastic cell; next generation; novel; novel strategies; novel therapeutics; oncolytic herpes simplex virus; oncolytic virotherapy; particle; receptor; receptor binding; trafficking; tumor; tumor eradication; tumor microenvironment; Address; Animals; Binding; Biology; Cell Adhesion Molecules; Cell fusion; Cell physiology; Cells; Complex; Creativeness; Cytolysis; Data; Diagnosis; Dose; E-Cadherin; Eating; Engineering; FDA approved; Fc Immunoglobulins; Glioblastoma; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Herpesvirus 1; Immune; Immune response; Immunity; Immunocompetent; Immunoglobulin G; Immunoglobulins; Immunologic Deficiency Syndromes; Immunosuppression; Infection; Innate Immune Response; Killer Cells; Laboratories; Ligands; Malignant Neoplasms; Manuscripts; Mediating; Minor; Modeling; Mus; N-Cadherin; NK Cell Activation; Names; Natural Killer Cells; Neoplasm Metastasis; Normal Cell; Oncolytic viruses; Operative Surgical Procedures; Paper; Patients; Penetration; Pharmaceutical Preparations; Play; Production; Publications; Publishing; Radiation therapy; Reporting; Reproduction; Research; Role; Signal Transduction; Site; Solid Neoplasm; Surface; Testing; Toxic effect; Viral; Virotherapy; Virus; anti-tumor immune response; blood-brain barrier crossing; cancer therapy; chemotherapy; chimeric antigen receptor; cytotoxicity; immune clearance; improved; in vivo; interest; neoplastic cell; next generation; novel; novel strategies; novel therapeutics; oncolytic herpes simplex virus; oncolytic virotherapy; particle; receptor; receptor binding; trafficking; tumor; tumor eradication; tumor microenvironment

Despite research efforts over the past two decades, glioblastoma (GBM) is still a devastating, deadly cancer with a median overall survival of less than 15 months. Oncolytic virotherapy is a promising approach for GBM treatment because it not only involves direct lysis of tumor cells and spares normal cells but also boosts immune responses to tumor cells. However, early clearance of oncolytic viruses by innate immune responses, poor virus propagation in tumor cells, and insufficient viral entry and/or spread present barriers that limit the efficacy of oncolytic virotherapy. In this application, we generated oncolytic herpes simplex virus 1 (oHSV) carrying a “don't eat me signal”, i.e., being engineered to express a ligand, E-cadherin, of the inhibitory receptor of natural killer (NK) cells, KLRG1. This next-generation oHSV carrying E-cadherin (named E-oHSV) reduces the deleterious NK-mediated clearance of oHSV. Importantly, unlike the drugs that we and others previously used to suppress oncolytic virus clearance by innate immune responses, the novel E-oHSV that we generated should not have an immunosuppressive effect systemically or even in the tumor (GBM) microenvironment. That is, E-oHSV only reduces activation of NK cells that bind to or attack E-oHSV-infected cells in the tumor microenvironment, while the remaining NK cells will maintain their potency against GBM cells. E-oHSV also showed enhanced viral production likely by increasing cell-to-cell fusion between virus- infected and uninfected cells. GBM cells do not have endogenous E-cadherin expression; however, interestingly, we found that E-cadherin is not only highly expressed on the surface of E-oHSV-infected cells but also loaded on free viral particles, the latter of which facilitates viral entry and/or spread in solid tumors. These improvements on oncolytic viruses led to significantly enhanced survival of mice bearing patient-derived GBM cells and also prolonged animal survival in an orthotopic, immunocompetent GBM model. In this application, we propose to mechanistically characterize this E-oHSV and to strengthen our hypothesis that a next- generation oHSV expressing E-cadherin, E-oHSV, plays multiple roles to overcome the barriers of oncolytic virotherapy and thus has a superior efficacy for GBM treatment. Three aims have been proposed: Aim 1 is to explore the mechanisms by which innate immune responses to E-oHSV are inhibited. Aim 2 is to explore the mechanisms by which E-oHSV enhances cell-to-cell infection, viral binding, and thus viral spread and distinguish them from the NK inhibition mechanism. Aim 3 is to investigate in vivo mechanisms, efficacy, and potential toxicity of E-oHSV for GBM treatment. Collectively, our study will not only address fundamental questions on NK cell and OV biology, but will also develop novel therapeutics for GBM treatment. It may also provide a platform to enhance efficacy of non-HSV oncolytic viruses.

尽管在过去二十年中进行了研究工作，但胶质母细胞瘤（GBM）仍然是一种毁灭性的致命癌症 总体生存率中位数不到15个月。溶瘤病毒疗法是GBM的一种有前途的方法 治疗是因为它不仅涉及肿瘤细胞的直接裂解并释放正常细胞，还涉及启动 对肿瘤细胞的免疫反应。但是，通过先天免疫反应对溶瘤病毒的早期清除， 肿瘤细胞中的病毒传播不良，病毒进入和/或传播障碍不足，以限制 溶瘤病毒疗法的功效。在此应用中，我们生成了溶瘤疱疹单纯疱疹病毒1（OHSV） 携带“不要吃我信号”，即正在设计以表达抑制性的配体e-钙粘着蛋白 天然杀手（NK）细胞的受体KLRG1。下一代OHSV携带E-钙粘蛋白（命名为E-OHSV） 减少了已删除的NK介导的OHSV的清除。重要的是，与我们和他人的药物不同 以前用来通过先天免疫反应抑制溶瘤病毒的清除，这是我们的新型E-OHSV 产生的应有系统甚至在肿瘤（GBM）中具有免疫抑制作用 微环境。也就是说，E-OHSV仅降低与感染E-OHSV感染的NK细胞的激活 肿瘤微环境中的细胞，而其余的NK细胞将保持其对GBM的效力 细胞。 E-OHSV还显示出可能通过增加病毒之间的细胞间融合而增强的病毒产生。 感染和未感染的细胞。 GBM细胞没有内源性E-钙粘蛋白表达。然而， 有趣的是，我们发现E-钙粘蛋白不仅在E-OHSV感染的细胞表面高度表达，而且还在 还加载了自由病毒颗粒，后者有助于病毒进入和/或扩散在实体瘤中。这些 改善溶瘤病毒导致带有患者衍生GBM的小鼠的生存率显着提高 细胞以及在原位，免疫功能的GBM模型中延长动物生存。在此应用程序中 我们建议机械地表征这一E-OHSV，并加强我们的假设，即接下来的假设 OHSV代表E-辅助蛋白E-OHSV扮演多个角色以克服障碍 溶瘤病毒疗法，因此具有较高的GBM治疗效率。三个目标是 提议：目标1是探索抑制先天免疫反应的机制。 AIM 2是探索E-OHSV增强细胞对细胞感染，病毒结合并因此病毒的机制 扩展并区分NK抑制机制。目标3是研究体内机制， E-OHSV对GBM治疗的疗效和潜在的毒性。总体而言，我们的研究不仅会解决 关于NK细胞和OV生物学的基本问题，但还将开发用于GBM治疗的新型疗法。 它还可以提供一个平台来提高非HSV溶瘤病毒的效率。