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

以 SOCS1 和 RASA2 为靶点，设计更有效的过继 T 细胞疗法来治疗癌症。

• 批准号: 10041283
• 负责人: JULIA Catherine CARNEVALE
• 金额: $ 26.33万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10041283
• 关键词: Adoptive Cell Transfers; Affect; Antigens; Antitumor Response; Area; Attenuated; Award; Basic Science; Biological; CAR T cell therapy; CD19 gene; CRISPR screen; CTAG1 gene; Cancer Center; Cancer Patient; Candidate Disease Gene; Cell physiology; Cells; Cellular biology; Characteristics; Chemicals; Clinic; Clinical; Clinical Trials; Combined Modality Therapy; Complement; Educational workshop; Electroporation; Engineering; Evaluation; Foundations; Funding; Gene Combinations; Gene Targeting; Genes; Genetic Transcription; Genome; Genome engineering; Goals; Human; Immune; Immune system; Immunocompetent; Immunodeficient Mouse; Immunomodulators; Immunotherapy; Implant; In Vitro; Infection; Knock-out; Laboratories; Libraries; MAP Kinase Gene; Malignant Neoplasms; Melanoma Cell; Membrane Proteins; Mentors; Mentorship; Methods; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; Oncology; Pathway interactions; Patients; Performance; Phenotype; Pre-Clinical Model; Property; Publishing; Research; Research Personnel; SILV gene; Signal Transduction; Solid Neoplasm; Spleen; Structure; System; T cell response; T cell therapy; T-Cell Activation; T-Cell Proliferation; T-Lymphocyte; Technology; Testing; Therapeutic; Training; Translational Research; Tumor Immunity; cancer cell; cancer immunotherapy; cancer therapy; career; cell behavior; cell killing; cellular transduction; effective therapy; engineered T cells; experience; gene discovery; genome wide screen; genome-wide; hands on research; immune checkpoint; improved; in vivo; in vivo Model; inhibitor/antagonist; interest; lead candidate; mouse model; neoplastic cell; next generation; next generation sequencing; novel; pre-clinical; preclinical evaluation; programs; research clinical testing; response; screening; single-cell RNA sequencing; skills; synergism; tool; tumor

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.

此K08应用程序的目标是为医学博士Julia Carnevale提供她将拥有的技能 需要成为一名独立资助的实验室研究员，研究设计T的新方法 具有更强抗肿瘤活性的细胞。临床上对改进有强烈的兴趣 采用T细胞疗法治疗和治愈更多癌症。然而，直到最近，还没有 有效和全面地评估基因组以找到关键的基因网络的方法 可以被用来编程有利的人类T细胞行为。卡内维尔博士最近研制出一种 用Cas9电穿孔(切片)筛选sgRNA慢病毒感染新技术 在原代人类T细胞中进行全基因组CRISPR筛查，以寻找调节关键基因 治疗功能。Carnevale博士使用切片对T细胞进行了全基因组筛查 发现了一系列增殖调节剂，其中许多还增强了T细胞的激活和 在体外杀死癌细胞。她现在建议从SOCS1这个屏幕中寻找排名靠前的基因 和RASA2(分别是JAK/STAT和MAP激酶信号的负调节因子)，以改善 过继T细胞疗法的疗效。她假设SOCS1或RASA2的中断将 增强人T细胞的抗肿瘤特性，即这些基因可以靶向改善 采用T细胞疗法，并可在体内进行切片联合筛查以确定 与SOCS1和RASA2缺失协同作用的修饰基因靶点，以改善肿瘤控制。这个 拟议研究的具体目的是：1)测试SOCS1和RASA2对T细胞的影响 对重复刺激的反应，并探索控制这些反应的分子途径 反应，2)确定SOCS1或RASA2失活对肿瘤细胞抗肿瘤活性的影响 3)在体内寻找与SOCS1或RASA2缺失协同作用的基因靶点。 卡内维尔博士的培训和研究计划包括有组织的课程作业和 研讨会、一对一教程和实践研究体验都将在 加州大学旧金山分校，世界著名的基础和翻译研究学术中心。 卡内维尔博士的培训计划将补充她现有的专业知识，为她奠定坚实的基础 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T细胞治疗的临床前模型，以及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·阿什沃斯博士的指导下进行， 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，以及HER 共同导师亚历克斯·马森博士，免疫细胞基因组工程方面的领先专家。她还会 从她杰出的咨询小组那里获得广泛的意见，并补充专业知识以指导 她的研究和职业轨迹。在完成这一奖项时，卡内维尔博士将获得 相关教学和研究经验，成为发现和发展的领导者 癌症患者的下一代采用T细胞疗法。