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: 10696150
• 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/10696150
• 关键词: Acute Myelocytic Leukemia; Adaptor Signaling Protein; 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; 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; Mentors; Mentorship; Non-Malignant; Normal tissue morphology; PTPN6 gene; Patients; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physicians; Predisposition; Protein Dephosphorylation; Protein Tyrosine Phosphatase; Proteomics; Receptor Activation; Receptor Signaling; Recurrence; 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; cancer cell; candidate identification; career development; chemotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; clinical translation; cytotoxicity; design; education planning; engineered T cells; experimental study; extracellular; high dimensionality; improved; in vitro Model; in vivo; innovation; interest; leukemia; leukemia initiating cell; leukemic stem cell; neoplastic cell; next generation; novel; preservation; protein expression; receptor; recruit; skills; success; therapeutic target; transcriptome sequencing; transcriptomics; transmission process; 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，美国有近20，000名患者被诊断患有急性髓性白血病（AML）， 5-尽管有高强度的化疗，在许多情况下， 移植嵌合抗原受体（CAR）T细胞治疗B细胞恶性肿瘤患者的成功， 激发了人们对将这种革命性的免疫疗法应用于AML的浓厚兴趣。但临床 翻译受到AML靶向表达在不可缺少的健康组织上的重叠的限制， 增加其对CAR T细胞介导的细胞毒性的易感性。内皮细胞毒性特别值得关注 一些正在调查的反洗钱目标为了推进AML的CAR T细胞治疗， 设计策略以保持抗白血病功效，同时使正常组织免于靶向， 肿瘤外毒性这一提议的中心假设是抗原特异性抑制性汽车（iCAR）可以是 经工程改造以调节CAR T细胞活化信号，并将降低AML CAR的靶向、脱瘤毒性 T细胞。在强有力的初步数据的指导下，这一假设将通过三个具体目标进行检验：1）确定 并入含有iCAR的NOT门控CAR T细胞的最佳抑制基序; 2）定义机制 iCAR内的那些抑制性基序可以利用先进的蛋白质组学技术中断CAR T细胞信号传导， 技术;和3）通过整合转录组学和细胞凋亡来确定内皮特异性NOT门表面靶标。 蛋白质组学数据。该提案的主要创新包括实施有针对性的筛选，以确定最佳的 类iCAR，将在靶点之间推广，并应用质谱细胞术（CyTOF）来询问 CAR T细胞中的基本抑制信号介体。虽然该提案的直接重点是设计 AML的非门控CAR，这些实验定义的原则将提供应用此方法的框架。 技术广泛应用于其他肿瘤类型。 拟议的研究活动是一项全面的职业发展计划的一部分， 申请人的专业知识，是至关重要的，她的发展作为一个独立的调查员，重点是 转译免疫疗法特别是，申请人将获得高维蛋白质组学的专业知识， 利用斯坦福大学世界一流的科学和导师环境， 大学申请人将由Crystal Mackall博士指导，她以基础CAR工作而闻名 T细胞免疫学和翻译免疫疗法，并由Ravi Majeti博士共同指导， AML干细胞的治疗靶向。在他们的指导下，沿着聚集的咨询委员会 （包括卡拉·戴维斯博士、安苏曼·萨特帕蒂博士和肯尼思·温伯格博士）和具体的教育计划， 申请人将获得所需的支持和资源，以实现拟议的目标，并有效地 在K 08培训期结束后向独立过渡。

项目成果

Development of clinical pathways to improve multidisciplinary care of high-risk pediatric oncology patients.

• DOI: 10.3389/fonc.2022.1033993
• 发表时间: 2022
• 期刊: Frontiers in oncology
• 影响因子: 4.7