NOT-gated CAR T cells to overcome on-target, off-tumor toxicity in AML

非门控 CAR T 细胞可克服 AML 中的靶向、肿瘤外毒性

• 批准号: 10524625
• 负责人: Rebecca Margaret Richards
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10524625
• 关键词: Acute Myelocytic Leukemia; Adaptor Signaling Protein; Address; Advisory Committees; Antigen Targeting; Antigens; B lymphoid malignancy; Bioinformatics; Biological Models; CAR T cell therapy; CD19 gene; Cell Line; Cell-Mediated Cytolysis; Cells; Cellular biology; Crystallization; Cytometry; Data; Data Set; Development; Development Plans; Diagnosis; Endothelial Cells; Endothelium; Engineering; Environment; Equilibrium; Event; Flow Cytometry; Foundations; Genetic Transcription; Goals; Green Fluorescent Proteins; Growth; Hematopoietic; Hematopoietic stem cells; Immune system; Immunohistochemistry; Immunology; Immunotherapy; Interruption; Investigation; Link; Logic; Malignant Neoplasms; Measures; Mediator of activation protein; Mentors; Mentorship; Non-Malignant; Normal tissue morphology; PTPN6 gene; Patients; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physicians; Predisposition; Protein Tyrosine Phosphatase; Proteomics; Receptor Activation; Receptor Signaling; Regulation; Reporter; Research Activity; Research Personnel; Resources; Safety; Sampling; Scientist; Signal Transduction; Signaling Molecule; Stem cell transplant; Structure; Surface; Survival Rate; T-Cell Activation; T-Cell Receptor; Techniques; Technology; Testing; Tissues; Toxic effect; Training; Transcriptional Activation; United States; Universities; Work; acute myeloid leukemia cell; acute toxicity; antileukemic activity; base; cancer cell; career development; chemotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; clinical translation; cytotoxicity; design; engineered T cells; experimental study; extracellular; high dimensionality; improved; in vitro Model; in vivo; innovation; interest; leukemia initiating cell; leukemic stem cell; neoplastic cell; next generation; novel; preservation; protein expression; receptor; recruit; skills; success; therapeutic target; transcriptome sequencing; transcriptomics; tumor; tumor specificity

PROJECT SUMMARY Each year, nearly 20,000 patients in the United States are diagnosed with acute myeloid leukemia (AML), and 5-year overall survival rates remain dismal despite high intensity chemotherapy and, in many cases, stem cell transplant. Success of chimeric antigen receptor (CAR) T cell therapy in patients with B cell malignancies has prompted intense interest in applying this revolutionary type of immunotherapy to AML. However, clinical translation has been limited by overlap of AML target expression on indispensable, healthy tissues, which increases their susceptibility to CAR T cell-mediated cytotoxicity. Endothelial cell toxicity is of particular concern for some AML targets under investigation. To advance CAR T cell therapy for AML, it is critically important to devise strategies to preserve anti-leukemic efficacy while simultaneously sparing normal tissue from on-target, off-tumor toxicity. The central hypothesis of this proposal is that antigen-specific inhibitory CARs (iCARs) can be engineered to modulate CAR T cell activation signals and will decrease on-target, off-tumor toxicity of AML CAR T cells. Guided by strong preliminary data, this hypothesis will be tested with three specific aims: 1) Identifying optimal inhibitory motifs to incorporate into iCAR-containing NOT-gated CAR T cells; 2) Defining mechanisms by which those inhibitory motifs within iCARs can interrupt CAR T cell signaling using advanced proteomic techniques; and 3) Determining endothelial-specific NOT-gate surface targets by integrating transcriptomic and proteomic data. Key innovations of this proposal include implementation of a targeted screen to identify a best- in-class iCAR that will be generalizable across targets and application of mass cytometry (CyTOF) to interrogate fundamental inhibitory signaling mediators in CAR T cells. While the immediate focus of this proposal is designing a NOT-gated CAR for AML, principles defined by these experiments will provide the framework for applying this technology broadly to other tumor types. The proposed research activities are part of a comprehensive career development plan that will build on past expertise of the applicant and are crucial to her development as an independent investigator focused on translational immunotherapy. In particular, the applicant will gain expertise in high dimensional proteomics and bioinformatics by taking advantage of the world class scientific and mentorship environment at Stanford University. The applicant will be mentored by Dr. Crystal Mackall, renowned for her work on fundamental CAR T cell immunology and translational immunotherapy, and co-mentored by Dr. Ravi Majeti, an expert on therapeutic targeting of AML stem cells. Under their guidance, along with the assembled advisory committee (including Drs. Kara Davis, Ansuman Satpathy, and Kenneth Weinberg) and specific educational plan, the applicant will receive the necessary support and resources to accomplish the proposed aims and efficiently transition to independence following the K08 training period.

项目摘要 每年，美国近20,000名患者被诊断出患有急性髓细胞性白血病（AML）和 尽管高强度化疗，但5年的总生存率仍然令人沮丧，在许多情况下，干细胞 移植。 B细胞恶性肿瘤患者的嵌合抗原受体（CAR）T细胞疗法的成功已有 对将这种革命性的免疫疗法应用于AML引起了强烈的兴趣。但是，临床 翻译受到AML目标表达在必不可少的健康组织上的重叠的限制， 增加了其对CAR T细胞介导的细胞毒性的敏感性。内皮细胞毒性特别关注 对于一些正在研究的AML目标。为了推进AML的CAR T细胞疗法，对 制定保留抗白血病功效的策略，同时从目标上避免正常组织 非肿瘤毒性。该提议的核心假设是抗原特异性抑制汽车（ICAR）可以是 设计用于调节CAR T细胞激活信号，并将减少AML CAR的靶向，肿瘤的毒性 T细胞。在强大的初步数据的指导下，该假设将以三个特定目的进行检验：1）确定 最佳的抑制基序将掺入含ICAR的未门控CAR T细胞中； 2）定义机制 通过高级蛋白质组学 技术； 3）通过整合转录组和 蛋白质组学数据。该提案的关键创新包括实施目标屏幕，以确定最佳 在跨目标和质量细胞术（Cytof）询问的跨越ICAR CAR T细胞中的基本抑制信号传导介质。虽然该提案的直接重点是设计 这些实验定义的原则将为应用程序提供框架 技术广泛地针对其他肿瘤类型。 拟议的研究活动是一项全面的职业发展计划的一部分，该计划将基于过去 申请人的专业知识，对于她作为专注于她的独立研究人员的发展至关重要 翻译免疫疗法。特别是，申请人将获得高维蛋白质组学的专业知识， 通过利用斯坦福大学的世界一流的科学和指导环境，生物信息学 大学。申请人将由Crystal Mackall博士指导，以其基本汽车的工作而闻名 T细胞免疫学和转化免疫疗法，并由Ravi Majeti博士合作 AML干细胞的治疗靶向。在他们的指导下，以及集会的咨询委员会 （包括卡拉·戴维斯（Kara Davis）博士，Ansuman Satpathy和Kenneth Weinberg）和特定的教育计划 申请人将获得必要的支持和资源，以实现拟议的目标并有效 在K08培训期间过渡到独立性。