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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要尽管 T 细胞介导的免疫反应对于免疫治疗的成功至关重要，但这些 T 细胞与恶性病变相关的通常是功能失调并且无法控制肿瘤生长。治疗用离体分离、激活和扩增的肿瘤浸润淋巴细胞 (TIL) 已被证明非常有效在一些黑色素瘤患者群体中。然而，相当多的患者没有反应，可能是由于许多宿主免疫因素之一。目前对 TIL 作用机制的了解表明肿瘤特异性效应 T 细胞的早期强劲扩增和低分化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 TIL 输注时高剂量外源性 IL-2，反应与高频率的相关性效应 T 细胞，以及大多数肿瘤杀伤发生在治疗开始后的早期。证据后者在许多临床前实验以及临床观察中发现，其中 TIL 的存在输注产品中的中央记忆子集与肿瘤消退相关。我们的总体目标是通过生成具有瞬时有效能力的 TIL 产品来提高 TIL 治疗效果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，我们开发了稳健的方法，用 mRNA 介导的基因重新编程 TIL 治疗。使用我们的mRNA方法具有安全性高、效率高、快速等优点生产、严格控制表达水平和同时多因子重编程。在初步在工作中，我们开发了一种系统，可以将 mRNA 的寿命延长一个数量级。我们的单细胞对扩增前和扩增后患者 TIL 的分析确定了扩增中缺乏的两个特定途径 TIL 产品可能导致其即时疗效不佳和记忆缺失的命运。这两个将通过 TIL mRNA 重编程得到增强。在目标 1 中，我们将开发一种增强扩展后的方法 TIL 生存，而在目标 2 中，我们将提高中央记忆 TIL 的产量。我们将评估效果使用源自多个黑色素瘤的配对肿瘤/TIL 组对 TIL 产生进行这些改进通过在体外和临床前人源化小鼠中研究肿瘤介导的 TIL 激活和肿瘤溶解来治疗患者使用单细胞分析和先进的空间转录组学的模型。该应用程序解决了以下需求提高黑色素瘤过继细胞免疫疗法的反应率，旨在可转化为在不久的将来进行临床试验。