Viscoelasticity and T Cell Production

虽然过继T细胞疗法(例如，抗CD19嵌合抗原受体(CAR)-T细胞)已经证明 白血病和淋巴瘤患者的显著疗效，在效力上仍存在显著差异 而且抗肿瘤反应的持久性，以及它们对抗实体瘤的成功一直是有限的。上一首 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细胞的产生通常需要体外T细胞激活和 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这一过程中被广泛决定。关于这个问题的重大研究 在过去的二十年里，细胞外基质(ECM)的弹性或硬度对许多方面都有影响 基本的细胞过程，并影响T细胞生物学的各个方面(例如，突触形成)。然而， 组织和细胞外基质不是线性弹性材料。ECM是粘弹性的，其对机械的响应 加载依赖于时间。基质粘弹性对干细胞分化的强烈影响 已证实，但基质刚性和粘弹性对T细胞激活的相互作用尚不清楚。这 该项目提出的假设是，激活过程中基质的粘弹性和刚性将直接影响T 细胞表型和治疗效果。这一假设将通过以下具体目标进行探讨：(1) 使用ECM模拟水凝胶评估基质粘弹性和硬度对T细胞表型的影响 利用可调刚度和粘弹性，(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患者分离的T细胞的关系 着重于AP-1途径，以及(3)阐明了T细胞状态变化对功能的影响 体内和体外基质力学采用采用转移研究。完成这些研究后， 提供有关基质力学在T细胞表型和功能上的作用的基础知识， 对为采用疗法制造T细胞的方法的潜在影响。

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.