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: 8897541
• 负责人: Rodger novak
• 金额: $ 28.73万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8897541
• 关键词: Address; Allogenic; Autologous; Autologous Transplantation; Berlin; Biodistribution; Biological Assay; Blood; Blood Cells; Bone Marrow Transplantation; CCR5 gene; CD4 Positive T Lymphocytes; Cell Count; Cells; Chemistry; Chemokine (C-C Motif) Receptor 5; 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; Work; Zinc Fingers; base; cell type; clinical application; clinically relevant; conditioning; cytotoxic; genome editing; graft vs host disease; in vivo; interest; manufacturing process; nuclease; null mutation; pre-clinical; prevent; receptor; stem; success; tool; treatment strategy

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.

这个合作项目的总体目标是制定一个“防御和摧毁”战略， HIV感染的功能性治愈，其中使患者的CD 4 + T细胞对 艾滋病毒感染（“防御”），同时采取战略消除病毒储存库 （“摧毁”）。这一战略的“防御”部分是基于“柏林病人”， 接受骨髓移植后，艾滋病毒感染治愈从一个捐助者窝藏无效 CCR 5基因突变，该基因编码HIV感染所必需的辅助受体。虽然 移植CCR 5无效造血干细胞（HSC）在理论上是一种有吸引力的方法。 HIV的治疗策略，实际上是从CCR 5缺失的异基因骨髓移植 由于供体有限，因此供体不是治疗普通人群中HIV的可行策略 可用性和与手术相关的风险，特别是移植物抗宿主病。在这里， 我们建议利用CRISPR-Cas9基因组编辑系统的力量， 患者自身造血干细胞（HSC）中的CCR 5突变，从而允许患者的造血干细胞（HSC）中的CCR 5突变。 自体移植和HIV的功能性治愈，而没有与自体移植相关的许多风险。 同种异体移植拟议战略的成功取决于最近 发现了CRISPR-Cas9基因组编辑系统，这是一个高度可定制的， 以高百分比的靶向突变CCR 5的两个拷贝所需的靶向效率 细胞群作为全面临床翻译《保卫与摧毁》的第一步 CRISPR Therapeutics Ltd公司是该应用的私营部门合作伙伴， 将在学术实验室开发的HSC中破坏CCR 5的过渡策略（见 项目1）通过执行两个具体目标实现临床应用：1） 开发符合适用法规的用于遗传修饰人HSC的方法 管理基于细胞的医药产品的法规;和2）药理学分析， 生物分布、毒理学和脱靶效应。的 这两个目标的实现将使该技术为首次人体临床试验做好准备， 并将为HIV的长期功能性治愈提供基础。更广泛地说，这些研究 将建立一个基本的临床准备平台，在此基础上， 例如无毒的预处理方案（项目3）和增强HSC归巢的策略， 可以增加病毒储存库的破坏（项目1和2）。更广泛地说， 在这里开发的用于在HSC中进行符合GMP的基因编辑的技术将具有广泛的应用前景。 适用于各种血液遗传性疾病。