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 CD19 嵌合抗原受体 (CAR)-T 细胞）已证明 白血病和淋巴瘤患者的疗效显着，但效力仍存在显着差异 抗肿瘤反应的持久性和持久性，并且它们对抗实体瘤的成功有限。以前的 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 CAR-T细胞输注产品中。 CAR-T细胞生产通常需要离体T细胞激活和 CAR-T 细胞输注产品的扩增和关键属性，包括其增殖能力， 持久性和抗肿瘤效力在此过程中广泛确定。重大研究 过去二十年已经证实细胞外基质 (ECM) 弹性或刚度影响许多 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， 组织和 ECM 不是线弹性材料。 ECM 是粘弹性的，它对机械响应 加载与时间相关。基质粘弹性对干细胞分化的强烈影响 已得到证实，但基质刚度和粘弹性对 T 细胞激活的相互作用尚不清楚。这 项目提出了这样的假设：激活期间的基质粘弹性和刚度将直接影响 T 细胞表型和治疗功效。该假设将通过以下具体目标进行探索：（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 具有可调的刚度和粘弹性，（2）探索基质力学调节T细胞的机制 激活过程中的分化及其与使用 scRNA-seq 分析从患者体内分离的 T 细胞的关系 重点关注 AP-1 途径，以及 (3) 阐明 T 细胞状态变化的功能效应 使用过继转移研究进行体外和体内基质力学。完成这些研究将 提供有关基质力学对 T 细胞表型和功能的作用的基础知识， 对用于过继疗法的 T 细胞制造方法的潜在影响。