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

该SBIR第二阶段拨款提案的总体目标是研究和开发GMP生产 用于实体瘤的下一代肿瘤浸润淋巴细胞（TIL）癌症治疗剂的方法。临床 在临床试验中，基于TIL的免疫疗法已经证明了肿瘤消退和增加的存活率， 不同的癌症类型。尽管这些令人鼓舞的临床结果，持久的抗肿瘤反应通常是 在一组患有晚期癌症如黑色素瘤的患者中观察到。这就强调了发展 更有效的基于TIL的策略，并扩大对不同癌症类型的疗效。TIL疗法是 部分受到肿瘤中TIL积累低、持久性低、T细胞受体（TCR）亲和力弱的阻碍 和/或亲合力以及细胞表面上肿瘤抗原的低表达。此外，存在 T细胞上的抑制性信号如Lag 3、Tim 3、CD 39，对产生有效的和 长期抗肿瘤T细胞反应。在我们已发表的和初步的研究中，激活MyD 88信号转导， 人或小鼠T细胞，增加抗肿瘤活性和抗肿瘤T细胞存活。通过融合高亲和力 CD 8 α（细胞外）到MyD 88（细胞内; CD 8 α：MyD 88），我们开发了一种新的通用平台， 其严格地在TCR与肿瘤细胞上的MHC抗原接合时激活MyD 88信号传导。 TILs细胞中的CD 8 α：MyD 88表达显著增加了对弱和次优水平的CD 8 α：MyD 88的应答。 肿瘤抗原，包括新抗原，并在初步研究中已经证明了非凡的性质， 减少T细胞进入“耗尽状态”。重要的是，使用CD 8 α共受体代表了 这是一种增强T细胞反应的“通用”方法，无论患者的病情如何， HLA型。通过SBIR第二阶段的应用，我们提出了以下目标。目标1将研究， 开发和生产GMP级g-逆转录病毒载体。在这里，我们将产生高病毒滴度生产 CD 8a：MyD 88-EGFRt PG-13细胞系，伴随分析测定，并生产GMP CD 8a：MyD 88-EGFRt g-逆转录病毒载体。为此，我们正在与商业合同合作 开发生产组织（CDMO），Vigene Biosciences生产GMP病毒载体。在Aim中 2、拟开发生产GMP CD 8a：MYD 88工程TIL（coR 8：AMPTM TIL）。在 与科罗拉多大学的盖茨制造工厂（GBF）合作 校园（UCAMC），我们将进行过程和分析检测，QC放行检测，开发和制造 GMP级coR 8：AMPTM TILs。这些目标的成功完成将导致pre-IND和IND申请 与FDA合作进行免疫难治性实体瘤患者的I期临床试验。这项活动将 与UCAMC的细胞治疗操作计划（CTOP）合作进行，盖茨 我们的团队成员在细胞治疗药物方面拥有丰富的经验， 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.