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，遭受了约100,000名美国人（3）， 〜6000名加利福尼亚人，全球数十万（4）。它通常会缩短寿命 即使拥有最佳医疗服务，几十年。 SCD是由β-珠蛋白基因中的单个突变引起的。 编辑可以直接纠正突变。我们已经开发了一种使用一种自体干细胞产品 CRISPR/CAS9核酸酶刺激血液干细胞中镰状突变的修复。可重复编辑 基于临床经验的校正水平将足以实现治疗作用。我们 提出完成该疗法的临床前开发的建议，从而导致IND申请 患有严重SCD的成年人的阶段临床试验。我们在2018年10月30日举行了一次预定会议， 要求CIRM资助以下活动，以支持与FDA讨论的IND提交： 1。证明在GMP条件下以临床规模生产最终细胞产品的能力。 我们将在CGMP下至少生成3个临床规模的批次，以符合所有发布标准 HSPC。我们还将完成一项药物稳定性研究。这些报告将完成 IND的CMC部分。 2。完成严格的遗传毒性评估。支持人类的鼠定征学模型 造血细胞将用于对制造的最终细胞产物进行毒理学研究 CGMP条件。长期植入后，将对小鼠进行评估以证明人类恶性的证据 使用组织病理学，流式细胞仪和基因组读数的细胞。 （预性问题8）。 3。完成其他临床前研究以建立产品安全性和效力。详尽 对最终制造试剂的脱靶基因组修饰的询问将解决 潜在的遗传毒性。我们还将评估HBB内目标网站上的大量删除和易位， 由编辑程序引起的HBB突变的性质和潜在影响，以及证据 可以反映早期肿瘤进展的克隆扩张（预性问题5,11）。 4.用最终的临床试验设计草稿和提交IND。我们将生成最终的临床协议，同意 形式，数据和安全管理计划，以支持ID的安全性和安全性临床试验和 患有严重SCD的成年人的可行性。我们将提交这些要求的监管（例如IRB，IBC，NIH RAC）审查，等待IND的最终批准。 这种疗法有可能通过产生一种治疗疗法来改变SCD的护理 适用于任何SCD患者，并且比同种异体造血干细胞移植（HSCT），更安全 因此，可以防止SCD并发症发生不可逆的损害。