Curing Sickle Cell Disease with CRISPR-Cas9 genome editing

利用 CRISPR-Cas9 基因组编辑治疗镰状细胞病

• 批准号: 10264257
• 负责人: Mark C Walters
• 金额: $ 47.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-12 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10264257
• 关键词: Address; Adult; Allogenic; American; Autologous; CRISPR/Cas technology; Caring; Cells; Clinical; Clinical Protocols; Clinical Trials Design; Clonal Expansion; Consent Forms; Cyclic GMP; Engraftment; Flow Cytometry; Funding; Genes; Genetic Diseases; Genomics; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Histopathology; Human; Induced Mutation; Institutional Review Boards; Longevity; Malignant - descriptor; Medical; Modification; Mus; Mutation; Nature; Patients; Pharmaceutical Preparations; Phase I Clinical Trials; Procedures; Reagent; Reporting; Safety; Safety Management; Sickle Cell Anemia; Site; Toxicology; United States National Institutes of Health; Xenograft Model; base; beta Globin; curative treatments; data management; early phase clinical trial; exhaustion; experience; genome editing; genotoxicity; meetings; nuclease; preclinical development; preclinical study; prevent; recessive genetic trait; repaired; safety and feasibility; sickling; stem cells; therapy development; tumor progression

Sickle Cell Disease (SCD) is a devastating recessive genetic disorder, afflicting ~100,000 Americans (3), ~6000 Californians, and hundreds of thousands more worldwide (4). It typically shortens lifespan by decades even with optimal medical care. SCD is caused by a single mutation in the ß-globin gene; gene editing can directly correct the mutation. We have developed an autologous stem cell product that uses a CRISPR/Cas9 nuclease to stimulate repair of the sickle mutation in blood stem cells. Editing reproducibly yields levels of correction that, based on clinical experience, will be sufficient for a curative effect. We propose to complete preclinical development of this therapy, leading to an IND application for an early phase clinical trial in adults with severe SCD. We held a pre-IND meeting on October 30, 2018, and request CIRM funding for the following activities to support IND submission as discussed with the FDA: 1. Demonstrate the capacity to manufacture final cell product under GMP conditions at clinical scale. We will generate at least 3 clinical-scale lots under cGMP that meet all release criteria, in plerixaformobilized HSPCs. We will also complete a drug product stability study. These reports will complete the CMC section of the IND. 2. Complete a rigorous genotoxicity assessment. A murine xenograft model that supports human hematopoietic cells will be used to perform a toxicology study on final cell product manufactured under cGMP conditions. After long-term engraftment, mice will be assessed for evidence of malignant human cells using histopathology, flow cytometry, and genomic readouts. (pre-IND Question 8). 3. Complete additional preclinical studies to establish product safety and potency. An exhaustive interrogation for off-target genomic modifications with the final manufacturing reagents will address potential genotoxicity. We will also assess large deletions and translocations at the HBB on-target site, the nature and potential impact of HBB mutations induced by the editing procedure, and evidence of clonal expansion that could reflect early-stage neoplastic progression (pre-IND Questions 5,11). 4. Draft and file an IND with a final clinical trial design. We will generate a final clinical protocol, consent form, and data and safety management plan to support the IND for a Phase I clinical trial of safety and feasibility in adults with severe SCD. We will submit these for required regulatory (e.g. IRB, IBC, NIH RAC) review, pending final approval of IND. This therapy has the potential to transform the care of SCD by producing a curative treatment that is applicable to any SCD patient and safer than allogeneic hematopoietic stem cell transplantation (HSCT), thus making it possible to prevent complications of SCD before they have done irreversible damage.

镰状细胞病 (SCD) 是一种毁灭性的隐性遗传疾病，困扰着约 100,000 名美国人 (3)， 约 6000 名加州人以及全世界数十万人的寿命通常会缩短 (4)。 即使在最佳的医疗护理下，SCD 也是由 β-珠蛋白基因的单一突变引起的； 编辑可以直接纠正突变。我们开发了一种使用自体干细胞产品。 CRISPR/Cas9 核酸酶可重复性地刺激修复血液干细胞中的镰状突变。 根据临床经验，产生的校正水平足以产生疗效。 提议完成该疗法的临床前开发，从而获得早期 IND 申请 我们于 2018 年 10 月 30 日举行了 IND 前会议，并在成人严重 SCD 中进行了临床试验。 请求 CIRM 为以下活动提供资金，以支持与 FDA 讨论的 IND 提交： 1. 展示在临床规模的 GMP 条件下生产最终细胞产品的能力。 我们将根据 cGMP 生产至少 3 个符合所有放行标准的临床规模批次，采用 Plerixaformobilized 我们还将完成药品稳定性研究。 IND 的 CMC 部分。 2. 完成严格的遗传毒性评估，支持人类的小鼠异种移植模型。 造血细胞将用于对生产的最终细胞产品进行毒理学研究 长期植入后，将评估小鼠的 cGMP 条件是否存在恶性人类的证据。 使用组织病理学、流式细胞术和基因组读数对细胞进行分析（IND 前问题 8）。 3. 完成额外的临床前研究以确定产品的安全性和效力。 使用最终生产试剂询问脱靶基因组修饰将解决 我们还将评估 HBB 靶位点的大量缺失和易位， 编辑程序引起的 HBB 突变的性质和潜在影响，以及证据 克隆扩增可以反映早期肿瘤进展（IND 前问题 5,11）。 4. 起草并提交包含最终临床试验设计的 IND 我们将生成最终临床方案、同意书。 表格、数据和安全管理计划，以支持 IND 进行安全性和安全性的 I 期临床试验 我们将提交这些资料以供监管机构（例如 IRB、IBC、NIH）使用。 RAC）审查，等待 IND 最终批准。 这种疗法有可能通过产生一种治愈性治疗来改变 SCD 的护理 适用于任何SCD患者，比异基因造血干细胞移植（HSCT）更安全， 因此可以在 SCD 并发症造成不可逆转的损害之前预防它们。