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: 10006816
• 负责人: Katy Rezvani
• 金额: $ 23.22万
• 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/10006816
• 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; 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.

摘要： 在过去的几年里，免疫效应细胞工程取得了巨大的进步，比如 癌症治疗。然而，嵌合抗原受体(CAR)修饰的T细胞有许多局限性。 为每个患者生产一种自体产品在逻辑上是繁琐和受限的 用于广泛的临床应用。汽车T细胞的制造通常需要几个星期的时间，这使得它不切实际 适用于疾病进展迅速的患者。此外，并不总是可能产生与临床相关的 来自大量预治疗的CAR T细胞的剂量，通常是淋巴细胞减少的患者。以前收集的同种异体 产品可以克服这些限制；然而，同种异体T细胞(即使是HLA相合的)携带 移植物抗宿主病的显著风险由其固有的αβT细胞受体抑制介导 它们作为临床产品使用，不需要进一步的操作来消除T细胞受体。自然杀手(NK) 细胞为汽车工程提供了一种非常有吸引力的T细胞替代品。NK细胞不会引起移植物抗宿主病 从而打开了生产现成产品的机会，供立即临床使用。此外，由于 基因工程的NK细胞也应该保留它们的全部天然受体，它们有可能发挥作用 通过CAR的特异性以外的机制产生的细胞毒性，原则上可以 减少由CAR靶向抗原丢失所介导的复发风险，正如CAR-T细胞疗法所报道的那样。 自体NK细胞可在体外复制生成，但其抗病毒活性极其有限。 汽车工程无法攻克的自体肿瘤。脐带血(CB)是一种很容易获得的 具有明显优势的同种异体NK细胞来源。CB以现成的冷冻产品形式提供， 这一优势已经得到了从 体外冷冻的CB单位。大容量冰冻脐血细胞转化为CAR转导NK细胞的研究 全球可转债银行库存有望实现广泛的可扩展性，这是个人无法复制的 需要筛查和白细胞分离的成年捐赠者。 在目标1中，我们将进行第一次非人类临床试验，以测试改造的CB-NK细胞的安全性和有效性。 表达抗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 扩散和持久性，并表达基于IC9的自杀基因，以解决安全问题 与直接毒性的潜在风险有关；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 使用一种新的逆转录病毒构建物从转化生长因子-β/SMAD信号轴转导NK细胞，除了 CD19和IL-15，包括显性阴性的人转化生长因子β受体II的基因(转化生长因子β- DNRII)用于下一代临床研究。