Project 4: Off-the-shelf engineered cord blood-derived natural killer cells for the treatment acute lymphoblastic leukemia

项目 4：现成的工程化脐带血自然杀伤细胞，用于治疗急性淋巴细胞白血病

• 批准号: 10931069
• 负责人: Katy Rezvani
• 金额: $ 29.34万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10931069
• 关键词: Acute Lymphocytic Leukemia; Adoptive Cell Transfers; Adoptive Transfer; Adult; Advanced Malignant Neoplasm; Advisory Committees; Allogenic; Antigen Targeting; Antigens; Autologous; B lymphoid malignancy; B-Lymphocytes; Blood Banks; CAR T cell therapy; CASP9 gene; CD19 gene; CD3 Antigens; Cancer Center; Cancer Patient; Cell Therapy; Cell physiology; Cells; Clinic; Clinical; Clinical Protocols; Clinical Research; Clinical Trials; Correlative Study; Data; Disadvantaged; Disease; Doctor of Medicine; Dominant Genes; Dominant-Negative Mutation; Dose; Effector Cell; Engineering; Equipment and supply inventories; Exhibits; Freezing; Generations; Genetic Engineering; Human; In Vitro; Individual; Innate Immune System; Institutional Review Boards; Interleukin-15; Laboratories; Leukapheresis; Lymphoid; Malignant Neoplasms; Measures; Mediating; Methods; Modality; Mus; Natural Killer Cell toxicity; Natural Killer Cells; Patient-Focused Outcomes; Patients; Positioning Attribute; Proliferating; Protocols documentation; Recombinant DNA; Refractory; Regulation; Relapse; Reporting; Reproducibility; Research; Retroviral Vector; Risk; Risk Reduction; Safety; Sampling; Series; Signal Transduction; Source; Specificity; T-Cell Receptor; T-Lymphocyte; Testing; Texas; Therapeutic; Toxic effect; Transforming Growth Factor beta; Umbilical Cord Blood; Universities; acute lymphoblastic leukemia cell; advanced disease; alpha-beta T-Cell Receptor; antileukemic activity; cancer cell; cancer therapy; cellular transduction; chimeric antigen receptor; chimeric antigen receptor T cells; chimeric gene; clinically relevant; conventional therapy; cost effective; cytokine; cytokine release syndrome; cytotoxicity; engineered NK cell; first-in-human; graft vs host disease; immunoengineering; immunotherapy clinical trials; improved outcome; in vivo; individual patient; innovation; leukemia; manufacture; next generation; novel; novel strategies; pharmacologic; pre-clinical; preclinical study; receptor; reconstitution; refractory cancer; relapse risk; response; safety testing; screening; suicide gene; tumor; tumor-immune system interactions; vector

Summary: The past several years have seen tremendous advances in the engineering of immune effector cells as therapy for cancer. However, chimeric antigen receptor (CAR)-modified T-cells have a number of limitations. The generation of an autologous product for each individual patient is logistically cumbersome and restrictive for widespread clinical use. The manufacturing of CAR T-cells often takes several weeks, making it impractical for patients with rapidly advancing disease. Furthermore, it is not always possible to generate clinically relevant doses of CAR T-cells from heavily pre-treated, often lymphopenic patients. A previously collected allogeneic product could overcome these limitations; however, allogeneic T-cells (even if HLA-matched) carry a significant risk of graft-versus-host disease (GVHD) mediated through their native αβ T-cell receptor prohibiting their use as a clinical product without further manipulation to eliminate the T cell receptor. Natural killer (NK) cells provide an extremely attractive alternative to T-cells for CAR engineering. NK cells do not cause GVHD and thus open opportunities to produce an off-the-shelf product for immediate clinical use. Moreover, as engineered NK cells should also retain their full array of native receptors, they have the potential to exert cytotoxicity through mechanisms other than that dictated by the specificity of the CAR, which in principle could reduce the risk of relapse mediated by loss of CAR-targeted antigen, as reported for CAR-T cell therapy. Autologous NK cells can be reproducibly generated in vitro, but have extremely limited activity against autologous tumor which cannot be overcome by CAR engineering. Cord blood (CB) is a readily available source of allogeneic NK cells with clear advantages. CB is available as an off-the-shelf frozen product, an advantage that has been bolstered by methods to generate large numbers of highly functional NK cells from frozen CB units ex vivo. The generation of CAR-transduced NK cells from frozen CB units stored in large global CB bank inventories holds promise for widespread scalability that cannot be replicated with individual adult donors who require screening and leukapheresis. In Aim 1 we will perform the first-inhuman clinical trial to test the safety and efficacy of CB-NK cells engineered to express a CAR against CD19 (a B cell-specific antigen), to ectopically produce IL-15 to support their in vivo proliferation and persistence, and to express a suicide gene, based on IC9, that will address safety concerns related to the potential risk of direct toxicity; In Aim 2 we will apply highly innovative correlative studies to describe the therapeutic potential of the clinical trial; Aim 3 in preclinical murine studies, we will protect the transduced NK cells from the TGF-β/SMAD signaling axis using a novel retroviral construct that, in addition to CAR.CD19 and IL-15, includes the gene for the dominant-negative version of human TGFβ receptor II (TGFβ- DNRII) for next-generation clinical studies.

概括： 在过去的几年中 癌症治疗。但是，嵌合抗原受体（CAR）修饰的T细胞具有许多局限性。 每位患者的自体产品的产生在逻辑上很麻烦且限制性 用于宽度临床使用。汽车T细胞的制造通常需要数周，使其变得不切实际 适用于快速前进的疾病的患者。此外，并非总是有可能产生与临床相关的 来自经过大量预处理的，通常是淋巴细胞减少患者的汽车T细胞剂量。先前收集的同种异体 产品可以克服这些局限性；但是，同种异体T细胞（即使HLA匹配）携带 禁止通过其天然αβT细胞受体介导的移植物抗宿主病（GVHD）的显着风险 它们用作临床产品，而无需进一步操纵以消除T细胞受体。天然杀手（NK） 细胞为汽车工程的T细胞提供了一种极具吸引力的替代品。 NK细胞不会引起GVHD 因此，开放了生产现成产品供临床使用的机会。而且， 工程的NK单元还应保留其全部本机接收器，它们有可能执行 通过除了汽车特异性所决定的机制，其细胞毒性原理可以 如CAR-T细胞疗法所报道，降低了因靶向汽车靶向抗原而介导的中继风险。 自体NK细胞可以在体外生成可重复产生，但具有极为有限的活性 汽车工程无法克服的自体肿瘤。脐带血（CB）很容易获得 具有明显优势的同种异体NK细胞的来源。 CB可作为现成的冷冻产品，一种 通过方法来产生大量高功能NK细胞的方法，从 冷冻的CB单元在体内。来自存储在大型的冷冻CB单元中的CAR转导的NK细胞的产生 全球CB银行库存具有无法与个人复制的宽度可伸缩性有望 需要筛查和白细胞术的成年捐助者。 在AIM 1中，我们将执行第一个人口临床试验，以测试CB-NK细胞设计的安全性和效率 要对CD19（A B细胞特异性抗原）表达汽车，以生态产生IL-15以支持其体内 基于IC9的扩散和持久性，并表达自杀基因，该基因将解决安全问题 与直接毒性的潜在风险有关；在AIM 2中，我们将应用高度创新的相关研究 描述临床试验的治疗潜力；目标3在临床前鼠研究中，我们将保护 使用新型逆转录病毒构建体从TGF-β/Smad信号轴翻译NK细胞，此外 CAR.CD19和IL-15包括人类TGFβ受体II的显性阴性版本（TGFβ-）的基因 DNRII）用于下一代临床研究。