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细胞重定向到他们的癌症， 是治疗难治性和难治性癌症的非常有前途的疗法。虽然CAR-T细胞疗法已经被广泛应用， 在治疗难治性B细胞癌方面非常成功，1，2，当定向治疗时， 更复杂的实体瘤和液体瘤，如多发性骨髓瘤。CAR T细胞面临的挑战包括： 免疫抑制性肿瘤微环境，其中通常表达抑制性配体，营养素和 氧气缺乏，细胞因子或可溶性因子分泌支持恶性细胞生长。 环境以及细胞内在缺陷可导致CAR T细胞增殖减少，功能抑制， 以及CAR T细胞持久性的丧失。先前开发的合成Notch（synNotch）受体允许控制 4，5，6 SynNotch T细胞可以递送用户定义的细胞内在或 外源性有效载荷，例如转录因子、细胞因子或抗原特异性的抗体，空间上 控制的方式。虽然这些受体回路可以用来解决复杂的神经系统的许多挑战， 在肿瘤中，这些受体缺乏启动细胞毒性的能力，如汽车或TCR。成功的， 持久的工程化T细胞需要识别，激活和杀死肿瘤细胞的能力，以及 能够产生有效载荷，以应对挑战性肿瘤中遇到的各种挑战 微环境最近，我设计了一种新的“杂交SynNotch CAR”受体，其结构 包含信号传导结构域（例如共刺激、CD 3 z等）可以启动T细胞的激活， 具有SynNotch受体的典型定制转录调节。这些杂交synNotch CAR受体是 功能和代表一类新的合成受体，激活短时间尺度的信号和长期的 在单一受体结构中的定制转录反应。本建议旨在扩大这些 最初的概念验证杂交SynNotch CAR受体，了解它们的全方位功能和作用， 工程化的T细胞，并证明其解决多发性骨髓瘤治疗挑战的能力。 加州大学旧金山分校弗朗西斯科（UCSF）是免疫学的领先机构，被认为是 作为生物医学科学研究生教育的顶级博士课程之一。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，并有机会获得 由帕克癌症免疫治疗研究所和Chan Zuckerberg Biohub提供的资源。这 培训计划还包括对专业发展计划的承诺，如UCSF培训计划 导师计划，以及手稿和论文写作课程。该提案旨在制定和应用一项 新型合成受体回路，它同时靶向癌细胞并解决了癌症治疗中的各种挑战。 肿瘤和肿瘤微环境。完成拟议的工作将推动工程癌症领域 免疫疗法向前发展，并为我未来的职业生涯做好准备，作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.