Project 2: Towards a safe and effective AML treatment strategy using anti-CD33 CAR T cells in combination with CAR-resistant hematopoietic stem cells

摘要/摘要(项目2) 免疫疗法使多种晚期恶性肿瘤的治疗发生了革命性变化。抗CD19嵌合体 抗原受体重定向T细胞(CART-19)在B细胞恶性肿瘤中尤其成功。如何 将CART细胞治疗的成功转化为其他恶性肿瘤，如急性髓系白血病(AML) 仍然是该领域的一个重要问题。CART细胞疗法的一个关键要求是靶组织 成为牺牲品。AML是一种造血干/祖细胞(HSPC)和SHARE细胞的恶性肿瘤 具有正常HSPC和正常髓系后代的表面抗原，如中性粒细胞和单核细胞。它 已经很明显，缺乏AML特异性抗原是释放AML的最大障碍 CART细胞对抗急性髓系白血病和其他髓系恶性肿瘤的能力。这个项目的长期目标是 为AML开发临床可行的CART细胞平台，方法是创建AML特定的CART细胞，该细胞能够 在体内扩大并持续存在，以根除AML，同时保留正常的骨髓功能。中心假说 抗CD33 CART细胞的持久抗白血病作用可以与足够水平的 基因工程CD33缺陷的造血。这将通过三个具体目标来实现。在AIM 1、研制高质量的可耗竭抗CD33 CAR T细胞。这些细胞将是同种异体的，供体来源的 用编码人源化抗CD33-41BB-Zeta CAR的双顺反子慢病毒载体转导T细胞 以及CD20。在目标2中，将测试制造CD33缺陷型人HSPC的可行性和安全性 生产CD33缺陷HSPC的监管部门将获得批准。在目标3中，将进行一项临床试验 进行了一种联合方法，其中包括CD33缺陷的、对汽车有抵抗力的异基因红细胞移植，然后 CART-33在急性髓系白血病患者中的应用这项研究将具有重要意义，因为它将有助于深度( 临床反应)和(有资格接受潜在根治疗法)的广度 AML。这项拟议研究的创新之处在于取代了寻找合适的白血病特异性基因 具有结合泛髓系特异性CART(如CART-33)和输液的新型平台的抗原 供者HSPC的基因工程缺乏CD33，因此对 Cart-33。

SUMMARY/ABSTRACT (PROJECT 2) Immunotherapy has revolutionized the treatment of a variety of advanced malignancies. Anti-CD19 chimeric antigen receptor redirected T cells (CART-19) have been particularly successful in B-cell malignancies. How to translate the success of CART cell therapy to other malignancies such as acute myeloid leukemia (AML) remains an important question in the field. A critical requirement of CART cell therapy is that the target tissue be expendable. AML is a malignancy of the hematopoietic stem/progenitor cells (HSPC) and shares cell surface antigens with normal HSPC and with normal myeloid progeny such as neutrophils and monocytes. It has become clear that the lack of AML-specific antigens is the single biggest impediment to unleashing the power of CART cells against AML and other myeloid malignancies. The long-term goal of this project is to develop a clinically feasible CART cell platform for AML, by creating AML-specific CART cells that are able to expand and persist in vivo to eradicate AML while preserving normal marrow function. The central hypothesis is that a durable anti-leukemic effect from anti-CD33 CART cells can co-exist with adequate levels of genetically engineered CD33-deficient hematopoiesis. This will be accomplished in three specific aims. In Aim 1, high quality depletable anti-CD33 CAR T cells will be manufactured. These will be allogeneic, donor-derived T cells transduced with a biscistronic lentiviral vector that encodes a humanized anti-CD33-41BB-zeta CAR as well as CD20. In Aim 2, the feasibility and safety of manufacturing CD33-deficient human HSPC will be tested and regulatory approvals to manufacture CD33-deficient HSPC will be obtained. In Aim 3, a clinical trial will be conducted of a combined approach incorporating CD33-deficient, CAR-resistant allogeneic HCT followed by CART-33 infusion in patients with AML. This research will be significant because it will contribute depth (of clinical responses) and breadth (of eligibility for potentially curative therapy) to the therapeutic arsenal against AML. The innovation of the proposed research lies in replacing the search for suitable leukemia-specific antigens with a novel platform that combines pan-myeloid specific CART (such as CART-33) with an infusion of donor HSPC that are genetically engineered to lack CD33 and which are therefore resistant to killing by CART-33.