Preclinical development of gene-edited human hematopoietic stem cells

基因编辑人类造血干细胞的临床前开发

• 批准号: 9052832
• 负责人: Rodger novak
• 金额: $ 27万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9052832
• 关键词: Address; Allogenic; Autologous; Autologous Transplantation; Berlin; Biodistribution; Biological Assay; Blood; Blood Cells; Bone Marrow Transplantation; CCR5 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Count; Cells; Chemistry; Clinic; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Development; Disease; Dose; Elements; Ensure; European; Gene-Modified; General Population; Genes; Genetic; Goals; Guide RNA; HIV; HIV Infections; Hematological Disease; Hematopoietic; Hematopoietic stem cells; Hereditary Disease; Homing; Homologous Transplantation; Human; Infection; Laboratories; Laboratory Research; Mediating; Methods; Modification; Mutate; Pathway interactions; Patients; Pharmacology; Population; Pre-Clinical Model; Private Sector; Procedures; Process; RNA analysis; Regimen; Regulation; Resistance; Risk; Safety; Source; Stem cells; System; Technology; Testing; Therapeutic; Toxic effect; Toxicology; Transfection; Translating; Translations; Transplantation; Tumorigenicity; Viral reservoir; Work; base; cell type; clinical application; clinically relevant; conditioning; cytotoxic; genome editing; graft vs host disease; in vivo; interest; manufacturing process; null mutation; pre-clinical; prevent; receptor; stem; success; tool; treatment strategy; zinc finger nuclease

The overall goal of this collaborative project is to develop a “Defend and Destroy” strategy for the functional cure of HIV infection in which a patient's CD4+ T cells are rendered resistant to HIV infection (“Defend”) while simultaneously strategies are applied to eliminate viral reservoirs (“Destroy”). The “Defend” portion of this strategy is based upon the “Berlin Patient,” who was cured of HIV infection upon receipt of a bone marrow transplant from a donor harboring a null mutation in the CCR5 gene, which encodes a co-receptor essential for HIV infection. Although transplantation with CCR5 null hematopoietic stem cells (HSCs) is theoretically an attractive treatment strategy for HIV, in practice allogeneic bone marrow transplants from CCR5-null donors are not a viable strategy for treating HIV in the general population due to limited donor availability and the risks associated with the procedure, particularly graft vs. host disease. Here, we propose to harness the power of the CRISPR-Cas9 genome editing system to introduce null mutations in CCR5 in a patient's own hematopoietic stem cells (HSCs), thereby allowing for an autologous transplant and a functional cure of HIV without many of the risks associated with an allogeneic transplant. The success of the proposed strategy hinges upon the recently discovered CRISPR-Cas9 genome editing system, which is a highly customizable and has the on-target efficiency needed to mutate both copies of CCR5 in a high percentage of the target cell population. As the first step towards full clinical translation of the “Defend and Destroy” strategy, the company CRISPR Therapeutics Ltd, the private-sector partner in this application, will transition strategies for disrupting CCR5 in HSCs developed in the academic laboratory (see Project 1) towards clinical application through the execution of two specific aims: 1) Development of a process for genetically modifying human HSCs compliant with applicable regulations governing cell-based medicinal products; and 2) Analysis of pharmacology, biodistribution, toxicology and off-target effects of genetically modified HSCs. The accomplishment of these two aims will ready this technology for a first-in-humans clinical trial, and will provide the basis for a long-term functional cure of HIV. More broadly, these studies will establish a basic clinic-ready platform upon which other elements of the overall strategy such as non-toxic conditioning regimens (Project 3) and strategies to enhance HSC homing or destruction of viral reservoirs (Projects 1 and 2) can be added. More broadly, the methods developed here for conducting GMP-compliant gene editing in HSCs will have widespread applicability to a variety of genetic diseases of the blood.

该合作项目的总体目标是制定“防御和摧毁”战略 HIV 感染的功能性治愈，患者的 CD4+ T 细胞对 HIV 感染具有抵抗力 HIV 感染（“防御”），同时应用策略消除病毒库 （“摧毁”）该策略的“防御”部分是基于“柏林病人”，他是 接受携带无效艾滋病毒的捐赠者的骨髓移植后，艾滋病毒感染被治愈 CCR5 基因突变，该基因编码 HIV 感染所必需的辅助受体。 CCR5 缺失的造血干细胞 (HSC) 移植理论上是一种有吸引力的方法 HIV 治疗策略，实践中 CCR5 缺失的同种异体骨髓移植 由于捐赠者有限，捐赠者并不是治疗普通人群艾滋病毒的可行策略 可用性以及与手术相关的风险，特别是移植物抗宿主病。 我们建议利用 CRISPR-Cas9 基因组编辑系统的力量来引入 null 患者自身造血干细胞 (HSC) 中的 CCR5 发生突变，从而允许 自体移植和艾滋病毒的功能性治愈，没有许多与移植相关的风险 同种异体移植的成功取决于最近的进展。 发现了 CRISPR-Cas9 基因组编辑系统，这是一个高度可定制的并且具有 在目标的高比例中突变 CCR5 的两个副本所需的达标效率 作为“防御与毁灭”全面临床转化的第一步。 战略，CRISPR Therapeutics Ltd 公司是本申请的私营部门合作伙伴， 将转变学术实验室开发的破坏 HSC 中 CCR5 的策略（参见 项目 1) 通过执行两个具体目标实现临床应用：1) 开发符合适用标准的人类 HSC 基因改造工艺 管理细胞医药产品的法规；以及 2) 药理学分析， 转基因 HSC 的生物分布、毒理学和脱靶效应。 这两个目标的实现将为这项技术进行首次人体临床试验做好准备， 更广泛地说，这些研究将为艾滋病毒的长期功能性治愈提供基础。 将建立一个基本的临床就绪平台，在此基础上总体战略的其他要素 例如无毒调理方案（项目 3）和增强 HSC 归巢的策略或 更广泛地说，可以添加破坏病毒库（项目 1 和 2）的方法。 这里开发的用于在 HSC 中进行符合 GMP 的基因编辑的技术将得到广泛应用 适用于多种遗传性血液病。