Viscoelasticity and T Cell Production

粘弹性和 T 细胞生产

• 批准号: 10566883
• 负责人: David J Mooney
• 金额: $ 56.89万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10566883
• 关键词: Address; Adoptive Transfer; Animals; Antitumor Response; CD19 gene; Cancer Patient; Cell Culture Techniques; Cell Separation; Cell physiology; Cellular biology; Dose; Elasticity; Environment; Extracellular Matrix; Gel; Hydrogels; Immune response; In Vitro; Infusion procedures; Knowledge; Malignant Neoplasms; Mechanics; Mediating; Outcome; Pathway interactions; Patients; Phenotype; Physical environment; Process; Production; Property; Research; Role; Signal Transduction; Solid Neoplasm; T cell differentiation; T cell therapy; T-Cell Activation; T-Lymphocyte; Therapeutic; Time; Tissues; Transcription Factor AP-1; Treatment Efficacy; anti-cancer; cancer immunotherapy; chimeric antigen receptor T cells; cytokine; imprint; improved; in vivo; knowledge of results; leukemia/lymphoma; manufacture; mechanical load; mimetics; physical property; response; single-cell RNA sequencing; stem cell differentiation; success; synaptogenesis; tumor; validation studies; viscoelasticity

While adoptive T cell therapies (e.g., anti-CD19 chimeric antigen receptor (CAR)-T cells) have demonstrated remarkable outcomes in patients with leukemias and lymphomas, significant variability remains in the potency and durability of the antitumor response, and their success against solid tumors has been limited. Previous studies have identified several key determinants of therapeutic efficacy, including distinct T-cell subpopulations in the CAR-T cell infusion product. CAR-T cell production generally requires ex vivo T-cell activation and expansion, and critical attributes of the CAR-T cell infusion product, including its proliferative capacity, persistence, and antitumor potency, are widely determined during this process. Significant research over the past two decades has established that extracellular matrix (ECM) elasticity, or stiffness, impacts many fundamental cell processes, and impacts various aspects of T cell biology (e.g., synapse formation). However, tissues and ECMs are not linearly elastic materials. The ECM is viscoelastic, with its response to mechanical loading being time dependent. Strong effects of matrix viscoelasticity on stem cell differentiation have been demonstrated, but the interplay of matrix stiffness and viscoelasticity on T cell activation is unknown. This project addresses the hypothesis that matrix viscoelasticity and stiffness during activation will directly impact T cell phenotype and therapeutic efficacy. This hypothesis will be explored via the following specific aims: (1) Assess the effects of matrix viscoelasticity and stiffness on T cell phenotype using ECM mimetic hydrogels with tunable stiffness and viscoelasticity, (2) Explore the mechanism by which matrix mechanics regulate T cell differentiation during activation, and its relation to T cells isolated from patients using scRNA-seq analysis and focusing on the AP-1 pathway, and (3) Elucidate the functional effects of changes in T cell state induced by matrix mechanics both in vitro and in vivo using adoptive transfer studies. Completion of these studies will provide fundamental knowledge regarding the role of matrix mechanics on T cell phenotype and function, with a potential impact on approaches to manufacture T cells for adoptive therapies.

虽然过继性T细胞疗法（例如，抗-CD 19嵌合抗原受体（CAR）-T细胞）已经证明 在白血病和淋巴瘤患者中的显著结果， 和抗肿瘤反应的持久性，并且它们对实体瘤的成功是有限的。先前 研究已经确定了治疗效果的几个关键决定因素，包括不同的T细胞亚群 在CAR-T细胞输注产品中。CAR-T细胞产生通常需要离体T细胞活化， 扩增和CAR-T细胞输注产品的关键属性，包括其增殖能力， 持久性和抗肿瘤效力在此过程中被广泛确定。在这方面的重要研究 过去二十年已经确定，细胞外基质（ECM）的弹性或刚度影响许多细胞外基质（ECM）。 基本细胞过程，并影响T细胞生物学的各个方面（例如，突触形成）。然而，在这方面， 组织和ECM不是线性弹性材料。ECM是粘弹性的，其响应于机械应力。 加载依赖于时间。基质粘弹性对干细胞分化的强烈影响已经被证实。 证明，但基质刚度和粘弹性对T细胞活化的相互作用是未知的。这 该项目解决了激活过程中基质粘弹性和刚度将直接影响T 细胞表型和治疗功效。本文将通过以下具体目标来探讨这一假设：（1） 使用ECM模拟水凝胶评估基质粘弹性和刚度对T细胞表型的影响 （2）探讨基质力学对T细胞的调控机制 活化期间的分化，以及其与使用scRNA-seq分析从患者分离的T细胞的关系， 聚焦于AP-1通路，和（3）阐明由AP-1通路诱导的T细胞状态变化的功能效应。 使用过继转移研究的体外和体内基质力学。完成这些研究将 提供关于基质力学对T细胞表型和功能的作用的基础知识， 对制造用于过继治疗的T细胞的方法的潜在影响。