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: 10247506
• 负责人: Katy Rezvani
• 金额: $ 27.24万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-05 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10247506
• 关键词: Acute Lymphocytic Leukemia; Address; 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; Cancer Center; Cancer Patient; Cell Therapy; Cell physiology; Cells; Clinic; Clinical; Clinical Data; 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; Logistics; Lymphoid; Malignant Neoplasms; Measures; Mediating; Methods; Modality; Mus; Natural Killer Cell toxicity; Natural Killer Cells; Patient-Focused Outcomes; Patients; Pharmacology; Positioning Attribute; Proliferating; Protocols documentation; Recombinant DNA; Refractory; Regulation; Relapse; Reporting; Reproducibility; Research; Retroviral Vector; Risk; 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; base; 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; graft vs host disease; immunoengineering; immunotherapy clinical trials; improved outcome; in vivo; individual patient; innovation; leukemia; next generation; novel; novel strategies; 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： 过去几年，免疫效应细胞工程取得了巨大进展， 然而，嵌合抗原受体（CAR）修饰的 T 细胞有许多局限性。 为每个患者生产自体产品在后勤上是麻烦且限制性的 CAR T 细胞的制造通常需要数周时间，因此不切实际。 此外，对于疾病快速进展的患者来说，并不总是能够产生临床相关的结果。 来自预先接受过大量治疗、通常患有淋巴细胞减少症的患者的 CAR T 细胞剂量 A 先前收集的同种异体。 产品可以克服这些限制；然而，同种异体 T 细胞（即使 HLA 匹配）携带 通过其天然 αβ T 细胞受体抑制介导的移植物抗宿主病 (GVHD) 的显着风险 它们作为临床产品使用，无需进一步操作即可消除 T 细胞受体 (NK)。 NK 细胞为 CAR 工程提供了一种极具吸引力的 T 细胞替代方案，不会引起 GVHD。 从而为生产可立即用于临床的现成产品提供了机会。 工程化 NK 细胞还应保留其全部天然受体，它们有潜力发挥作用 通过 CAR 特异性以外的机制产生细胞毒性，原则上可以 根据 CAR-T 细胞疗法的报道，降低因 CAR 靶向抗原丢失而介导的复发风险。 自体 NK 细胞可以在体外重复生成，但其抗病毒活性极其有限 无法通过 CAR 工程克服的自体肿瘤 脐带血 (CB) 是一种现成的方法。 CB 是一种具有明显优势的同种异体 NK 细胞来源，可作为现成的冷冻产品使用。 产生大量高功能 NK 细胞的方法增强了这一优势 体外冷冻 CB 单位 从储存在大容量冷冻 CB 单位中生成 CAR 转导的 NK 细胞。 全球 CB 银行库存有望实现广泛的可扩展性，而个人无法复制 需要筛查和白细胞分离术的成年捐献者。 在目标 1 中，我们将进行首次人体临床试验，以测试工程化 CB-NK 细胞的安全性和有效性 表达针对 CD19（一种 B 细胞特异性抗原）的 CAR，异位产生 IL-15 以支持其体内 增殖和持久性，并表达基于 IC9 的自杀基因，这将解决安全问题 与直接毒性的潜在风险相关；在目标 2 中，我们将应用高度创新的相关研究来 描述临床试验的治疗潜力；目标 3 在临床前小鼠研究中，我们将保护 使用新型逆转录病毒构建体从 TGF-β/SMAD 信号轴转导 NK 细胞，除了 CAR.CD19 和 IL-15，包括人 TGFβ 受体 II 显性失活版本的基因 (TGFβ- DNRII）用于下一代临床研究。