Enhancing Melanoma TIL Efficacy with Multifactor mRNA-Mediated T Cell Reprogramming

通过多因子 mRNA 介导的 T 细胞重编程增强黑色素瘤 TIL 功效

• 批准号: 10721549
• 负责人: MICHAEL E HURWITZ
• 金额: $ 23.49万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10721549
• 关键词: Acclimatization; Address; Adoptive Immunotherapy; Antitumor Response; Apoptotic; Back; Cell Differentiation process; Cell Reprogramming; Cell Therapy; Cells; Clinical; Clinical Trials; Cytolysis; Data; Dose; Electroporation; Environment; Fishes; Fluorescent in Situ Hybridization; Frequencies; Future; Generations; Genes; Goals; Harvest; Image; Immune checkpoint inhibitor; Immune system; Immunologic Factors; Immunotherapy; In Vitro; In complete remission; Individual; Infrastructure; Infusion procedures; Insertional Mutagenesis; Interleukin-2; Knowledge; Laboratories; Lesion; Ligands; Longevity; Lymphocyte; Lymphocyte Activation; Malignant - descriptor; Malignant Neoplasms; Mediating; Memory; Messenger RNA; Metastatic Melanoma; Methods; OX40; Outcome; Pathway interactions; Patients; Phenotype; Population; Population Heterogeneity; Positioning Attribute; Production; Property; Proteins; Protocols documentation; RNA; Rejuvenation; Research; Safety; Sampling; System; T cell receptor repertoire sequencing; T cell therapy; T-Cell Receptor; T-Lymphocyte; TNFSF4 gene; Therapy Clinical Trials; Time; Transfection; Treatment Efficacy; Tumor Antigens; Tumor Expansion; Tumor-Infiltrating Lymphocytes; Variant; Work; advanced disease; cell mediated immune response; cytokine; design; effector T cell; efficacy evaluation; experimental study; fighting; gene therapy; humanized mouse; improved; instrument; long term memory; lymphocyte product; mRNA Expression; manufacture; melanoma; mouse model; multidisciplinary; neoplastic cell; novel; objective response rate; patient derived xenograft model; patient population; pre-clinical; programs; response; single cell analysis; single-cell RNA sequencing; success; transcriptomics; tumor; tumor growth; tumor microenvironment

PROJECT SUMMARY Although T cell mediated immune responses are critical for the success of immunotherapy, those T cells associated with malignant lesions are typically dysfunctional and fail to control tumor growth. Treatment with tumor infiltrating lymphocytes (TIL) that are isolated, activated, and expanded ex vivo has proven very effective in some patient populations of melanoma. However, a substantial number of patients do not respond, presumably due to one of a number of host immune factors. Current understanding of TIL mechanism of action suggests that both an early robust expansion of tumor-specific effector T cells and transfer of less differentiated cells with long-term survival capacity are key to a successful therapy. Evidence for the former includes the need for high dose exogenous IL-2 at the time of TIL infusion, the correlation of response with a high frequency of effector T cells, and the majority of tumor killing occurring very early after the initiation of therapy. Evidence for the latter is found in many pre-clinical experiments as well as clinical observations where the presence of TILs from the central memory subset in the infusion product correlates with tumor regression. Our overall goal is to improve TIL therapeutic efficacy through the generation of TIL products with both the transient ability to effectively immediately kill tumor cells as well as the long-term ability to persist and maintain durable anti-tumor responses. To address these challenges we have developed robust methods to reprogram TILs with mRNA-mediated gene therapy. The use of our mRNA approach has the advantages of increased safety, high efficiency, rapid production, tightly controlled expression levels and simultaneous multi-factor reprogramming. In preliminary work we have developed a system that increases mRNA lifespan by an order of magnitude. Our single cell analysis of patient TILs pre- and post-expansion has identified two specific pathways deficient in the expanded TIL product that likely contribute to their poor immediate efficacy and absence of memory fate. Both of these will be augmented by TIL mRNA reprogramming. In Aim 1 we will develop a method to enhance post-expansion TIL survival, while in Aim 2 we will improve the production of central memory TILs. We will evaluate the effect of these improvements in TIL production using paired tumor/TIL sets derived from the melanomas from multiple patients by studying tumor-mediated TIL activation and tumor lysis in vitro and in pre-clinical humanized mouse models using single cell analysis and advanced spatial transcriptomics. This application addresses the need to improve response rates to adoptive cell immunotherapies for melanoma and is designed to be translatable to clinical trials in the near future.

项目摘要 尽管T细胞介导的免疫反应对于免疫疗法的成功至关重要，但这些T细胞 与恶性病变相关的通常是功能失调，无法控制肿瘤的生长。与 被证明非常有效 在某些患者黑色素瘤中。但是，大量患者没有反应， 大概是由于许多宿主免疫因子之一。当前对行动机制的理解 表明既早期肿瘤特异性效应T细胞的稳健膨胀，又是分化较小的转移 长期生存能力的细胞是成功治疗的关键。前者的证据包括需要 对于TIL输注时高剂量的外源IL-2，反应与高频的相关性 效应T细胞和大多数肿瘤杀伤发生在开始治疗后很早就发生。证据 后者在许多临床前实验以及临床观察中发现了TIL的存在 从输注产物中的中央记忆子集与肿瘤回归相关。我们的总体目标是 通过产生具有有效的瞬态能力的直到产物来提高治疗功效 立即杀死肿瘤细胞，以及持久和维持持久抗肿瘤反应的长期能力。 为了应对这些挑战，我们开发了可靠的方法来重新编程mRNA介导的基因 治疗。我们的mRNA方法的使用具有提高安全性，高效率，快速的优势 生产，紧密控制的表达水平和同时进行多因素重编程。在初步 我们已经开发了一个系统，该系统将mRNA寿命增加一个数量级。我们的单元 扩张前后患者tils的分析已经确定了两种缺乏扩展的特定途径 直到可能有助于其立即效力和缺乏记忆命运的产品。这两个 TIL mRNA重编程将增强。在AIM 1中，我们将开发一种增强扩张后的方法 直到生存期，在AIM 2中，我们将改善中央记忆的产生。我们将评估效果 在使用成对的肿瘤/TIL集中，这些改进的TIL产生源自来自多个黑色素瘤 通过研究肿瘤介导的TIL激活和肿瘤裂解的患者，并在临床前人性化小鼠中 使用单细胞分析和高级空间转录组学模型。此申请解决了需要 提高对黑色素瘤的收养细胞免疫疗法的反应率，并被设计为可翻译成 在不久的将来进行临床试验。