Next Generation Autologous TIL Cancer Therapy: Development of GMP manufacturing process

这项第二阶段SBIR资助计划的总体目标是研究和发展GMP制造 下一代肿瘤浸润性淋巴细胞(TIL)癌实体瘤的治疗进展。在临床上 试验表明，基于TIL的免疫疗法显示肿瘤消退并提高了存活率。 不同的癌症类型。尽管有这些令人鼓舞的临床结果，持久的抗肿瘤反应通常是 在黑色素瘤等晚期癌症患者中观察到。这强调了发展的必要性 更有效的基于TIL的策略，并扩大对不同类型癌症的疗效。TIL疗法是 部分原因是肿瘤中低TIL积聚、持续时间低、T细胞受体(TCR)亲和力弱 和/或亲和力以及肿瘤抗原在细胞表面的低表达。此外，存在的 T细胞上的抑制信号，如LAG3，TIM3，CD39，构成了产生有效和 长寿的抗肿瘤T细胞反应。在我们已发表的和初步的研究中，激活MyD88信号在 人或小鼠T细胞，增强抗肿瘤活性，延长T细胞存活时间。通过融合高亲和力 CD8α(细胞外)到MyD88(细胞内；CD8α：MyD88)我们开发了一个新颖的通用平台 这在TCR与肿瘤细胞上的MHC抗原结合时严格激活MyD88信号。 CD8α：MyD88在TIL细胞中的表达显著增强对弱和次适水平 肿瘤抗原包括新抗原，并在初步研究中表现出非凡的性质 减少T细胞进入“精疲力竭状态”。重要的是，CD8α共受体的使用代表了 一种增强T细胞反应的“通用”方法，可用于患者，而不考虑患者的 人类白细胞抗原类型。通过这项SBIR第二期申请，我们提出了以下目标。目标一号将进行研究， 研制GMP级g-逆转录病毒载体。在这里，我们将产生高滴度的病毒 CD8a：MyD88-EGFRt PG-13细胞株，附分析检测，并制作GMP CD8a：MyD88-EGFRt g-逆转录病毒载体。为此，我们与一家商业公司签订了合作合同 开发制造组织(CDMO)，Vigene Biosciences，以产生GMP病毒载体。在AIM 2、我们计划研制GMP CD8a：MyD88工程化TIL(CoR8：AMPTM TILs)。在……里面 与科罗拉多大学安舒茨医学院盖茨制造设施(GBF)的合作 校园(UCAMC)我们将进行过程分析测试，QC释放测试，开发和制造 GMP级CoR8：AMPTM瓷砖。这些目标的成功完成将导致IND预审和IND申请 与FDA合作，对免疫难治性实体肿瘤患者进行I期临床试验。这项活动将是 与UCAMC的细胞治疗操作计划(CTOP)合作进行，盖茨 生物制造设施，以及我们在细胞治疗药物方面拥有丰富经验的团队成员 发展。

The overall goal of this Phase II SBIR grant proposal is to research and develop the GMP manufacturing process for next-generation tumor infiltrating lymphocyte (TIL) cancer therapeutic for solid tumors. In clinical trials, TIL-based immunotherapies have demonstrated tumor regression and increased survival rates against different cancer types. Despite these encouraging clinical results, durable antitumor responses are typically observed in a subset of patients with advanced cancers like melanoma. This stresses the need to develop more effective TIL-based strategies and to expand efficacy against different cancer types. TIL therapies are hindered in part by low TIL accumulation into tumors, low persistence, weak T cell receptor (TCR) affinity and/or avidity as well as the low expression of tumor antigens on the cell’s surface. Additionally, the presence of suppressive signals on T cells such as Lag3, Tim3, CD39, pose a major obstacle to generating effective and long-lived antitumor T cell responses. In our published and preliminary studies, activating MyD88 signaling in human or mouse T cells, increases antitumor activity and prolongs T cell survival. By fusing the high affinity CD8α (extracellular) to MyD88 (intracellular; CD8α:MyD88) we have developed a novel and universal platform that activates MyD88 signaling strictly upon the engagement of the TCR with the MHC-antigen on tumor cells. CD8α:MyD88 expression in TILs cells substantially increases responses to weak and suboptimal levels of tumor antigens including neoantigens, and in preliminary studies has demonstrated the extraordinary property of reducing the T cells entry into an ‘exhausted state’. Importantly, the use of the CD8α co-receptor represents a ‘universal’ approach to enhancing T cell responses, and can be used in patients regardless of the patient’s HLA type. Through this SBIR Phase II application, we propose the following aims. Aim 1 will research, develop, and manufacture GMP grade g-retroviral vectors. Here, we will generate high virus titer-producing CD8a:MyD88-EGFRt PG-13 cell lines, accompanying analytical assays, and manufacture the GMP CD8a:MyD88-EGFRt g-retroviral vectors. For this aim, we are partnering with a commercial contract development manufacturing organization (CDMO), Vigene Biosciences to generate GMP viral vectors. In Aim 2, we propose to develop and manufacture GMP CD8a:MYD88 engineered TILs (coR8:AMPTM TILs). In collaboration with the Gates Manufacturing Facility (GBF) at the University of Colorado Anschutz Medical Campus (UCAMC) we will conduct process & analytical assay, QC release assay, develop and manufacture of GMP grade coR8:AMPTM TILs. The successful completion of these aims will lead to the pre-IND and IND filings with the FDA for a Phase I clinical trial in patients with immune refractory solid tumors. This activity will be conducted in collaboration with the UCAMC’s Cell Therapy Operations Program (CTOP), Gates Biomanufacturing Facility, and our team members who have extensive experience in cell therapy drug development.