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 虽然T细胞介导的免疫应答对于免疫治疗的成功至关重要，但这些T细胞介导的免疫应答对于免疫治疗的成功至关重要。 与恶性病变相关的肿瘤细胞通常功能失调并且不能控制肿瘤生长。治疗 已证明离体分离、活化和扩增的肿瘤浸润淋巴细胞（TIL）非常有效 在一些黑色素瘤患者群体中。然而，相当多的病人没有反应， 可能是由于宿主免疫因素之一。目前对TIL作用机制的理解 表明肿瘤特异性效应T细胞的早期稳健扩增和分化程度较低的细胞的转移， 具有长期存活能力的细胞是成功治疗的关键。前者的证据包括需要 对于在TIL输注时的高剂量外源性IL-2，应答与高频率的 效应T细胞，并且大多数肿瘤杀伤发生在治疗开始后非常早期。证据 后者在许多临床前实验以及临床观察中发现， 来自输注产品中的中央记忆子集的信息与肿瘤消退相关。我们的总体目标是 通过产生具有瞬时能力的TIL产物来提高TIL治疗功效， 立即杀死肿瘤细胞以及长期持续和维持持久抗肿瘤反应的能力。 为了应对这些挑战，我们已经开发了稳健的方法来用mRNA介导的基因重组TILs。 疗法使用我们的mRNA方法具有增加的安全性、高效性、快速性和特异性的优点。 生产，严格控制表达水平和同时多因子重编程。初步 我们已经开发出一种系统，可以将mRNA的寿命提高一个数量级。我们的单细胞 对患者TIL扩增前和扩增后的分析已经确定了扩增后的TIL中缺乏的两种特异性途径。 TIL产品可能导致其即刻疗效差和缺乏记忆结局。这两 将被TIL mRNA重编程增强。在目标1中，我们将开发一种方法来增强后扩展 TIL存活，而在目标2中，我们将提高中央记忆TIL的产生。我们将评估效果 在使用源自来自多个肿瘤的黑色素瘤的配对肿瘤/TIL组的TIL产生的这些改善中， 通过在体外和临床前人源化小鼠中研究肿瘤介导的TIL活化和肿瘤溶解， 使用单细胞分析和先进的空间转录组学模型。该应用程序满足了以下需求： 提高了对黑素瘤过继细胞免疫疗法的应答率，并被设计成可转化为 在不久的将来进行临床试验。