Multi-antigen-specific CAR T cells to treat acute myeloid leukemia

项目总结 嵌合抗原受体(CAR)T细胞疗法是一种新型的细胞免疫疗法，在这种疗法中 使用合成受体重定向T细胞的抗原特异性。CD19-CAR T细胞已完全达到 高达90%的急性淋巴细胞白血病患者的反应。然而，许多恶性肿瘤并不是 拥有单一的、高表达的肿瘤相关抗原(TAA)，如CD19。此外，CD19阴性 CD19-CAR T细胞治疗后复发频繁，提示多抗原- 将需要有针对性的方法来减少复发。急性髓系白血病(AML)是最常见的 成人的急性白血病和大多数患者将死于他们的疾病。我们和其他人正在评估 CAR T细胞治疗急性髓系白血病。然而，AML表现出TAA的异质性表达，并且许多这样的TAA 表达在造血祖细胞(HPC)上，增加了抗原阴性AML免疫的风险 AML靶向CAR T细胞治疗后的逃逸和骨髓毒性。此外，AML 采用许多主动免疫抑制策略，可能会抑制CAR T细胞。 为了克服这些挑战，我最近开发了一种新的病毒共转导和分选 允许产生和纯化具有多个转基因的T细胞的系统，例如多个CARS，免疫... 刺激分子、安全开关和分泌的细胞因子。初步数据显示，多功能 CAR T细胞可以被改造成克服抗原阴性白血病逃逸和免疫抑制 机械装置。我推测这种新的分选系统可以用来改造T细胞来克服急性髓系白血病 TAA异质性和免疫抑制策略。AIM 1将同时研究CAR T细胞 针对一组AML TAAs，并预测可避免对HPC的毒性。CAR T细胞被设计成克服AML- 还将对诱导免疫抑制进行评估。在目标2中，目标是针对一组由 AML和HPC都是移植前CAR T细胞免疫治疗策略的一部分。 在获奖期间，候选人将在纪念斯隆·凯特琳癌症中心进行研究 在马塞尔·范登布林克博士和一个咨询委员会的指导下。他将获得他所拥有的关键技能 需要成为一名终身教职的医生兼科学家，经营自己的学术实验室，开发合成 改进细胞疗法的生物学方法，并成功地竞争独立的NIH资金。他 将为同基因、异种移植和人源化小鼠模型培养详细而全面的技能 细胞免疫治疗，建立在现有的分子结构设计和细胞基因知识基础上 通过掌握多路CRISPR/Cas9基因中断和位点特异性基因整合进行修饰，以及 精通基因组分析，以更好地确定T细胞的激活和耗竭状态，并识别 基因治疗的新靶点。

PROJECT SUMMARY Chimeric antigen receptor (CAR) T cell therapy is a novel form of cellular immunotherapy in which the antigen specificity of T cells is redirected using synthetic receptors. CD19-CAR T cells have achieved complete responses in up to 90% of patients with acute lymphoblastic leukemia. However, many malignancies do not possess a single, highly expressed tumor-associated antigen (TAA) such as CD19. Furthermore, CD19-negative relapses have been frequently encountered following CD19-CAR T cell therapy, suggesting that multi-antigen- targeting approaches will be needed to reduce relapse. Acute myeloid leukemia (AML) is the most common acute leukemia in adults and the majority of patients will die from their disease. We and others are evaluating CAR T cells to treat AML. However, AML exhibits heterogeneous expression of TAAs and many of these TAAs are expressed on hematopoietic progenitor cells (HPCs), increasing the risk of antigen-negative AML immune escape and bone marrow toxicity following AML-targeting CAR T cell therapy, respectively. Additionally, AML employs many active immune-suppressive strategies that may inhibit CAR T cells. To overcome these challenges, I have recently developed a novel viral co-transduction and sorting system to allow generation and purification of T cells with multiple transgenes such as multiple CARs, immune- stimulating molecules, safety switches, and secreted cytokines. Preliminary data suggest that multi-functional CAR T cells can be engineered to overcome antigen-negative leukemia escape and immune suppression mechanisms. I hypothesize that this novel sorting system can be used to engineer T cells to overcome AML TAA heterogeneity and immune suppressive strategies. Aim 1 will investigate CAR T cells simultaneously targeting a set of AML TAAs and predicted to avoid toxicity to HPCs. CAR T cells engineered to overcome AML- induced immune suppression will also be evaluated. In Aim 2 the goal is to target a set of TAAs expressed by both AML and HPCs as part of a pre-transplant CAR T cell immunotherapy strategy. During the award period, the candidate will conduct research at Memorial Sloan Kettering Cancer Center under the mentorship of Dr. Marcel van den Brink and an Advisory Committee. He will obtain the critical skills he needs to become a tenure-track physician-scientist running his own academic laboratory developing synthetic biology approaches to improve cellular therapies and successfully competing for independent NIH funding. He will cultivate a detailed and comprehensive skill set for syngeneic, xenograft, and humanized mouse models of cellular immunotherapy, build upon an existing knowledge base of molecular construct design and cellular gene modification by mastering multiplexed CRISPR/Cas9 gene disruptions and site-specific gene integration, and develop proficiency in genomic analysis to better define T cell activation and exhaustion states and to identify novel targets for gene therapy.