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.

该协作项目的总体目标是制定“捍卫和破坏”策略 HIV感染的功能治愈，其中患者的CD4+ T细胞具有抗性 艾滋病毒感染（“防御”）同时采用策略来消除病毒库 （“破坏”）。该策略的“防御”部分是基于“柏林患者”的 从携带无效的捐赠者那里收到骨髓移植后治愈艾滋病毒感染 CCR5基因中的突变，该突变编码对HIV感染必不可少的共受体。虽然 用CCR5无效的造血干细胞（HSC）移植在理论上是一个吸引人的 艾滋病毒的治疗策略，实际上CCR5无效的同种异体骨髓移植 捐助者不是由于捐助者有限而导致普通人群治疗艾滋病毒的可行策略 可用性以及与该程序有关的风险，尤其是移植物与宿主疾病。这里， 我们建议利用CRISPR-CAS9基因组编辑系统的力量引入NULL 患者自身造血干细胞（HSC）中CCR5突变，从而允许 自体移植和艾滋病毒的功能治疗，没有许多与 同种异体移植。拟议策略的成功取决于最近 发现的CRISPR-CAS9基因组编辑系统，它是一种高度可定制的，具有 在高比例的目标中突变CCR5的两个副本所需的目标效率 细胞种群。作为迈向“防御和摧毁”完全临床翻译的第一步 策略，公司CRISPR Therapeutics Ltd，本申请中的私营部门合作伙伴， 将在学术实验室开发的HSC中破坏CCR5的过渡策略（请参阅 项目1）通过执行两个具体目标来临床应用：1） 开发一个符合适用的人类HSC的基因修饰的过程 管理基于细胞的医疗产品的法规； 2）药理学分析， 一般修饰的HSC的生物分布，毒理学和脱靶效应。这 实现这两个目标将为这项技术做好准备，以进行首个人类临床试验， 并将为艾滋病毒的长期功能治愈提供基础。更广泛地，这些研究 将建立一个基本的诊所就绪平台，在该平台上，整体战略的其他要素 例如无毒调节方案（项目3）和增强HSC归宿的策略或 可以添加病毒储层的破坏（项目1和2）。更广泛地，方法 在此开发用于在HSC中进行符合GMP兼容的基因编辑将具有宽度 适用于各种血液遗传疾病。