Investigating immunophenotype and metabolism of TCR KO donor and third-party CD19-targeted chimeric antigen receptor T cells

摘要 复发是异基因造血细胞移植（allo-HCT）后死亡的最重要原因，但 几十年来研究进展甚微。靶向CD 19（CD 19）的嵌合抗原受体 汽车）重定向T细胞效应子功能以消除表达CD 19的白血病和淋巴瘤细胞。然而，在这方面， 许多患者仍然复发。候选人有临床前数据表明使用基因组编辑的可行性 删除内源性T细胞受体（TCR）以降低供体CD 19 CAR T细胞的同种异体反应性，但事实确实如此 目前尚不清楚这些TCR敲除（KO）细胞在体内如何作为抗肿瘤剂发挥作用，移植到何种程度， 抗宿主病（GVHD）将导致，或工程如何影响T细胞代谢。这种知识是 对于为血液病患者创造易于获得的“现成”CAR T细胞的进展至关重要。 恶性肿瘤。拟议研究的总体目标是确定捐助者和第三方TCR KO CD 19 CAR T细胞在临床前模型中影响免疫重建、GVHD和移植物排斥，以及 了解去除TCR如何影响这些细胞的免疫代谢。核心假设是 有效的抗肿瘤作用以及可忽略的GVHD和移植物排斥反应可以在临床前证实 通过在allo-HCT后使用TCR KO CD 19 CAR T细胞（供体或第三方），以产生上级 在allo-HCT后，常规CD 19 CAR T细胞的结果。这一假设将在拟议的 具体目标。 这项工作将为候选人提供理想的培训，因为她为她的长期职业目标做好准备， 研究细胞疗法和allo-HCT的独立实验室。Memorial Sloan Kettering癌症中心 有一个著名的免疫学项目，候选人的主要导师马塞尔货车登布林克博士是一个领导者 免疫疗法研究的一部分。她的共同导师和咨询委员会成员具有多样性和互补性 他们都有指导独立科学家的良好记录。候选人和她 指导团队制定了严格的培训计划，旨在增加她的知识基础：1） 下一代CAR T细胞的开发; 2）生物信息学和编程; 3）细胞代谢， CAR T细胞中的代谢通量、线粒体功能和代谢分析;以及4）专业发展 skills.候选人将通过课程，研讨会和导师在这些领域进行培训。 这个研究项目和培训将为她未来的独立职业生涯奠定基础。 物理学家兼科学家预计拟议的研究将产生指导未来基因组的发现 CAR T细胞的工程化。候选人的目标是推出一个独立的研究计划，设计“关闭- “货架”CAR T细胞，预计将为一系列疾病提供更好的治疗选择。 血液恶性肿瘤。

Abstract Relapse is the most important cause of mortality after allogeneic hematopoietic cell transplant (allo-HCT), but little research progress has been made in several decades. Chimeric antigen receptors targeting CD19 (CD19 CARs) redirect T cell effector functions to eliminate CD19-expressing leukemia and lymphoma cells. However, many patients still relapse. The candidate has preclinical data indicating the feasibility of using genome editing to delete the endogenous T cell receptor (TCR) to reduce the alloreactivity of donor CD19 CAR T cells, but it is unknown how these TCR knockout (KO) cells will function in vivo as anti-tumor agents, to what extent graft- versus-host-disease (GVHD) will result, or how engineering impacts T cell metabolism. This knowledge is essential for progress toward creating readily available “off-the-shelf” CAR T cells for patients with hematologic malignancies. The overall objectives of the proposed research are to determine how donor and third-party TCR KO CD19 CAR T cells impact immune reconstitution, GVHD, and graft rejection in preclinical models, as well as to understand how removing the TCR impacts the immunometabolism of these cells. The central hypothesis is that potent anti-tumor effects as well as negligible GVHD and graft rejection can be demonstrated preclinically by using TCR KO CD19 CAR T cells (either donor or third-party) following allo-HCT, to produce superior outcomes to conventional CD19 CAR T cells following allo-HCT. This hypothesis will be tested in the proposed Specific Aims. This work will provide ideal training for the candidate as she prepares for her long-term career goal to lead an independent laboratory studying cellular therapeutics and allo-HCT. Memorial Sloan Kettering Cancer Center has a renowned immunology program, and Dr. Marcel van den Brink, the candidate’s primary mentor, is a leader in immunotherapy research. Her co-mentor and advisory committee members have diverse and complementary expertise, and all have strong track records of mentoring independent scientists. The candidate and her mentoring team have developed a rigorous training plan designed to increase her knowledge base in: 1) development of next-generation CAR T cells; 2) bioinformatics and programming; 3) cellular metabolism, metabolic flux, mitochondrial function, and metabolic analyses in CAR T cells; and 4) professional development skills. The candidate will undertake training in these areas through coursework, workshops, and mentorship. This research project and training will provide the foundation to establish her future career as an independent physician-scientist. The proposed studies are expected to generate findings that will guide future genome engineering of CAR T cells. The candidate’s aim is to launch an independent research program designing “off- the-shelf” CAR T cells, which are expected to provide much improved therapeutic options for a range of hematologic malignancies.