Targeting SOCS1 and RASA2 to Engineer More Potent Adoptive T Cell Therapies for Cancer Treatment.

这个K 08应用程序的目标是为博士提供朱莉娅·科瓦莱，医学博士，与她将 我需要成为一个独立资助的实验室研究员，研究新的方法来设计T 具有更强的抗肿瘤活性的细胞。有强烈的临床兴趣， 过继性T细胞疗法治疗和治愈更多癌症。然而，直到最近， 一种有效和全面评估基因组的方法，以找到关键的基因网络， 可以被用来编程有利的人类T细胞行为。Dr. Dallavale最近开发了一种 新的筛选技术，sgRNA慢病毒感染与CAS9电穿孔（SLICE）， 能够在原代人类T细胞中进行全基因组CRISPR筛选，以寻找调节关键基因的基因。 治疗功能。Dr. Kristivale使用SLICE在T细胞中进行全基因组筛选， 鉴定了一系列增殖调节因子，其中许多也能增强T细胞活化， 体外杀死癌细胞。她现在建议从这个筛选中靶向顶级基因SOCS 1 和RASA 2（分别为JAK/STAT和MAP激酶信号传导的负调节剂），以改善 过继性T细胞疗法的功效。她假设SOCS 1或RASA 2的破坏将导致 增强人类T细胞的抗肿瘤特性，这些基因可以靶向改善 过继性T细胞疗法，并且可以在体内进行SLICE合并筛选以鉴定 修饰基因靶点与SOCS 1和RASA 2丢失协同作用，以改善肿瘤控制。的 本研究的具体目的是：1）检测SOCS 1和RASA 2如何影响T细胞 对重复刺激的反应，并探索控制这些反应的分子途径。 2）确定SOCS 1或RASA 2失活对 体内T细胞，以及3）鉴定与体内SOCS 1或RASA 2丢失协同作用的基因靶点。 博士Eschervale的培训和研究计划包括结构化的课程和 研讨会，一对一的教程，和动手研究的经验，都将发生在 UCSF是世界著名的基础和转化研究卓越学术中心。 博士埃克塞特瓦莱的培训计划将补充她现有的专业知识，以建立一个强大的基础 在以下领域：1）基础T细胞生物学，包括调节电路的研究， 功能和肿瘤免疫，2）过继性T细胞疗法的临床前建模，和3） 下一代测序方法和分析，包括单细胞RNA测序。这个项目 将在她的主要导师，艾伦阿什沃思博士，总统的指导下进行。 加州大学旧金山分校癌症中心和治疗发现和转化研究的世界专家，她 Alex Marson博士是免疫细胞基因组工程领域的领先专家。她还将 从她杰出的顾问团获得广泛的投入，并提供互补的专业知识， 她的研究和职业轨迹在这个奖项完成后，Dr. 相关的教学和研究经验，成为发现和发展的领导者 癌症患者的下一代过继性T细胞疗法。

The goal of this K08 application is to provide Dr. Julia Carnevale, MD, with the skills she will need to become an independently-funded laboratory investigator studying novel ways to engineer T cells with more potent anti-tumor activity. There is intense clinical interest in improving adoptive T cell therapies to treat and cure more cancers. However, until recently there has been no way to efficiently and comprehensively evaluate the genome to find the critical gene networks that can be leveraged to program favorable human T cell behaviors. Dr. Carnevale recently developed a novel screening technology, sgRNA lentiviral infection with CAS9 electroporation (SLICE), that enables genome-wide CRISPR screening in primary human T cells to search for genes that regulate key therapeutic functions. Dr. Carnevale used SLICE to perform a genome-wide screen in T cells which identified a host of regulators of proliferation, many of which also enhanced T cell activation and in vitro cancer cell killing. She now proposes targeting top-raking genes from this screen, SOCS1 and RASA2 (negative regulators of JAK/STAT and MAP kinase signaling, respectively), to improve the efficacy of adoptive T cell therapies. She hypothesizes that disruption of SOCS1 or RASA2 will enhance the anti-tumor properties of human T cells, that these genes can be targeted to improve adoptive T cell therapies, and that SLICE pooled screening can be performed in vivo to identify modifier gene targets that synergize with SOCS1 and RASA2 loss to improve tumor control. The specific aims of the proposed research are: 1) to test how SOCS1 and RASA2 influence T-cell responses to repeated stimulation and explore the molecular pathways that govern these responses, 2) to determine the effects of SOCS1 or RASA2 inactivation on antitumor activities of T cells in vivo, and 3) to identify gene targets that synergize with SOCS1 or RASA2 loss in vivo. Dr. Carnevale's training and research plans includes a combination of structured coursework and workshops, one-on-one tutorials, and hands-on research experience that will all take place at UCSF, a world-renowned academic center of excellence in basic and translational research. Dr. Carnevale's training plan will complement her existing expertise to build a strong foundation in the following areas: 1) fundamental T cell biology including the study of regulatory circuitry, function, and tumor immunity, 2) preclinical modeling of adoptive T cell therapies, and 3) next-generation sequencing methods and analysis including single-cell RNA sequencing. This project will be conducted under the mentorship of her primary mentor, Dr. Alan Ashworth, President of the UCSF Cancer Center and world expert in therapeutic discovery and translational research, and her co-mentor, Dr. Alex Marson, leading expert in genome engineering in immune cells. She will also receive extensive input from her distinguished advisory panel with complementary expertise to guide her research and career trajectory. At the completion of this award, Dr. Carnevale will have the relevant didactic and research experience to become a leader in discovering and developing next-generation adoptive T cell therapies for cancer patients.