Dissecting and engineering CAR T-cell function for optimized Immunotherapy

项目总结 我的长期职业目标是为癌症研究开发可加速的翻译技术 从桌面到临床的发现对临床试验和患者管理产生了真正的影响。我的 目前的研究利用了生物材料、微系统和生物制造方面的工程进展，为新的 并改进了癌症生物学和免疫工程中新出现的问题的临床解决方案。特定的 例如用于单细胞传感和免疫监测的芯片实验室系统、胶质母细胞瘤、脑肿瘤 用于癌症快速诊断和预后的微环境建模，以及用于 探索干细胞和免疫细胞机械生物学。我提议在#年以新的身份扩大我的工作 用于现场免疫治疗患者筛查的新型工程系统的转化性癌症研究。 最近FDA批准嵌合抗原受体(CAR)T细胞免疫疗法用于B细胞 恶性肿瘤，CAR T细胞疗法是治疗复发和难治性白血病以及 实体瘤。然而，在许多临床试验中，CAR-T免疫疗法的临床益处差异很大 而且在复发/难治性白血病试验中报告的总体患者反应仍然是不利的。影响因素 导致不同的临床反应可能来自早期步骤，如CAR T细胞制造或 给药、CAR T细胞耗竭和免疫抵抗在白血病利基中的作用，但关键因素 导致CAR T细胞功效变化的原因尚不完全清楚。 我们研究的目标是开发新的工程系统来探索和分析 CAR T细胞的免疫学和生物力学特性及其白血病骨髓生态位的研究进展 目前的CAR T细胞免疫疗法。首先，我们的目标是重建一种新的器官型白血病BM 免疫生态位体外模型研究不同类型B细胞免疫抑制机制的异质性 各亚型及临床前评估和优化CD19车T细胞免疫治疗效果。其次，我们 目的开发和集成单细胞水平和(或)原位细胞和分子免疫表型系统 以便提供可靠和准确的筛选以表征功能 CAR T细胞的状态。最后，我们将探讨CAR-T细胞调节CAR-T的机械敏感机制。 在细胞活化和杀伤过程中提高CAR T细胞的效能。基于CAR T-CELL的新见解 机械生物学，我们的目标是设计一种远程“机械开关”，并结合一个“机械推进器”来 有效控制CAR T细胞活化和细胞毒作用，提高CAR T细胞免疫治疗效果 安全。总之，我们提出了一个创新的框架来精确地绘制时空免疫和 CAR T细胞激活和杀伤过程中的生物力学动力学研究 和机械签名的CAR T细胞，最终优化CAR T细胞的管理、安全和疗效。

PROJECT SUMMARY My long-term career goal is to develop translational technologies for cancer research that can accelerate discoveries from the benchtop to the clinic to make a real impact on clinical trials and patient management. My current research leverages engineering advances in biomaterials, microsystems, and biomanufacturing for new and improved clinical solutions to emerging problems in cancer biology and immune engineering. Specific examples include lab-on-a-chip systems for single-cell sensing and immunomonitoring, glioblastoma brain tumor microenvironment modeling for rapid cancer diagnosis and prognosis, and micromechanical systems for exploring stem and immune cell mechanobiology. I proposes to expand on my work in new capacity in translational cancer research for novel engineering systems for on-site immunotherapeutic patient screening. With the recent FDA approval of chimeric antigen receptor (CAR) T-cell immunotherapies for B-cell malignancies, CAR T-cell therapies are a promising strategy to cure relapsed and refractory leukemia as well as solid tumors. However, the clinical benefit of CAR-T immunotherapy varies tremendously in many clinical trials and overall patient responses reported in trials of relapsed/refractory leukemia remain unfavorable. Factors that contribute to variable clinical responses may arise from early steps like CAR T-cell manufacturing or administration, CAR T-cell exhaustion and immunological resistance in the leukemic niche, but the key elements leading to variations in CAR T-cell efficacy are not fully understood. The objective of our research is to develop novel engineering systems to probe and analyze both the immunological and biomechanical attributes of CAR T-cells and map the leukemic BM niche for advancing current CAR T-cell immunotherapies. First of all, we aim to reconstruct a novel organotypic leukemic BM immunity niche ex vivo model to dissect the heterogeneity of immunosuppression mechanisms of different B- ALL subtypes and pre-clinically evaluate and optimize CD19 CAR T-cell immunotherapy efficacy. Secondly, we aim to develop and integrate in situ cellular and molecular immunophenotyping systems at single-cell level and/or in a 3D organotypic setting so as to provide a reliable and accurate screening to characterize the functional status of CAR T-cells. Lastly, we will explore CAR T-cell mechanosensitive mechanisms that regulate CAR T- cell activation and killing process to improve the CAR T-cell efficacy. Based on the new insights from CAR T-cell mechanobiology, we aim to engineer a remote “mechanical switch” and incorporate a “mechanical promoter” to effectively control CAR T-cell activation and cytotoxicity for improved CAR T-cell immunotherapy efficacy and safety. Altogether, we propose an innovative framework to precisely map the spatiotemporal immunological and biomechanical dynamics during CAR T-cell activation and killing, aiming to construct ex vivo leukemic BM niche and mechanical signature of CAR T-cells, ultimately optimize CAR T-cell administration, safety, and efficacy.