Engineering Hybrid SynNotch CAR Receptors to Enhance Cell Therapies for Cancer

嵌合抗原T细胞(CAR-T)疗法，将患者自身的T细胞重新定向到他们的癌症， 是治疗难治和难治性癌症的非常有前途的疗法。虽然CAR T细胞疗法一直是 在治疗难治性B细胞癌方面非常成功，1，2他们在指导下遇到了许多挑战 更复杂的实体和液体肿瘤，如多发性骨髓瘤。CAR T细胞面临的挑战包括 免疫抑制的肿瘤微环境，抑制配体经常在其中表达，营养物质和 缺乏氧气，分泌细胞因子或可溶性因子以支持恶性细胞的生长。 环境以及细胞固有缺陷会导致CAR T细胞增殖减少、功能抑制 和丧失CAR T细胞持久性。先前开发的合成Notch(SynNotch)受体允许控制 和治疗性免疫细胞的定制。4，5，6 SynNotch T细胞可以传递用户定义的细胞固有或 外来有效载荷，如特定抗原中的转录因子、细胞因子或抗体，在空间上 控制的方式。虽然这些感受器电路可以用来解决复杂的 对于肿瘤，这些受体缺乏像CARS或TCRs那样启动细胞毒性的能力。因此，成功和 持久工程T细胞既需要识别、激活和杀伤肿瘤细胞的能力，也需要 能够产生有效载荷，以应对在挑战肿瘤时遇到的各种挑战 微环境。最近，我设计了一款新型的混合动力SynNotch汽车，它的结构 包含信号域(例如，共刺激、CD3z等)可以启动伴随的T细胞的激活 具有典型的SynNotch受体的定制转录调控。这些混合SynNotch汽车接收器是 具有功能，代表了一类新的合成受体，激活了短时间尺度信号和长时间 单一受体体系结构中的定制转录反应。这项提议试图在这些基础上进行扩展 初步概念验证混合SynNotch汽车受体，了解它们的全面功能和影响 设计了T细胞，并展示了它们解决多发性骨髓瘤治疗挑战的能力。 加州大学旧金山分校(UCSF)是免疫学领域的领先机构，被认为 作为生物医学科学研究生教育的顶尖博士项目之一。作为一名研究生，在 Roybal实验室，我定期接受知名研究人员的指导和指导，并可以访问 由帕克癌症免疫治疗研究所和陈·扎克伯格生物中心提供的资源。这 培训计划还包括对职业发展计划的承诺，如加州大学旧金山分校的培训计划 导师计划，以及手稿和论文写作课程。这项提案旨在开发和应用一种 新的合成受体电路，它同时靶向癌细胞并解决不同的挑战 肿瘤和肿瘤微环境。拟议工作的完成将推动工程癌症领域的发展 免疫疗法向前发展，并为我未来的职业生涯做好准备，成为一名独立的调查员。

Chimeric Antigen T Cell (CAR-T) therapies, which redirect a patient’s own T cells towards their cancer, are very promising therapies for difficult to treat and refractory cancers. While CAR T cell therapies have been very successful in treating refractory B cell cancers,1,2 they have encountered many challenges when directed towards more complex solid and liquid tumors, such as multiple myeloma. CAR T cells face challenges such as an immunosuppressive tumor microenvironment, in which inhibitory ligands are often expressed, nutrients and oxygen are lacking, and cytokines or soluble factors are secreted to support malignant cell growth.3 This hostile environment, as well as cell intrinsic defects, can lead to reduced CAR T cell proliferation, inhibition of function and loss of CAR T cell persistence. Previously developed synthetic Notch (synNotch) receptors allow for control and customization of therapeutic immune cells.4,5,6 SynNotch T cells can deliver user defined cell intrinsic or extrinsic payloads, such as transcription factors, cytokines, or antibodies in an antigen specific, spatially controlled manner. While these receptor circuits could be used to address the many challenges of complex tumors, these receptors lack the ability to initiate cytotoxicity like CARs or TCRs. Therefore, successful and persistent engineered T cells require both the ability to recognize, activate and kill tumor cells, as well as the ability to produce payloads to counteract a wide range of challenges encountered in challenging tumor microenvironments. Recently, I have engineered a novel “Hybrid SynNotch CAR” receptor, whose architecture incorporates signaling domains (e.g. co-stimulation, CD3z, etc.) that can initiate activation of T cells concomitant with custom transcriptional regulation typical of a SynNotch receptor. These Hybrid synNotch CAR receptors are functional and represent a new class of synthetic receptors that activate short timescale signaling and long term custom transcriptional responses in a single receptor architecture. This proposal seeks to expand upon these initial proof-of-concept Hybrid SynNotch CAR receptors, understanding their full range of function and effect on engineered T cells, and demonstrating their ability to address therapeutic challenges in multiple myeloma. The University of California, San Francisco (UCSF) is a leading institution in immunology and is regarded as one of the top doctoral programs in Biomedical Sciences graduate education. As a graduate student in the Roybal lab, I receive regular guidance and mentorship from renowned investigators and have access to resources provided by the Parker Institute for Cancer Immunotherapy, and the Chan Zuckerberg Biohub. This training plan also includes commitment to professional development programs, such as the UCSF TRAIN UP Mentorship program, and courses in manuscript and thesis writing. This proposal seeks to develop and apply a novel synthetic receptor circuit, which simultaneously targets cancer cells and addresses diverse challenges in tumors and tumor microenvironments. Completion of the proposed work will move the field of engineered cancer immunotherapy forward and prepare me for a future career as an independent investigator.