CCR5-mutant monkey model to facilitate the development of novel stem cell-based therapies for AIDS

CCR5突变猴模型促进新型干细胞艾滋病疗法的开发

• 批准号: 9140295
• 负责人: THADDEUS G GOLOS
• 金额: $ 76.44万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9140295
• 关键词: Acquired Immunodeficiency Syndrome; Address; Allogeneic Bone Marrow Transplantation; Allogenic; Animals; Anti-Retroviral Agents; Asians; Blood typing procedure; Bone Marrow Transplantation; CCR5 gene; CRISPR/Cas technology; CXCR4 gene; Cell Line; Cell Therapy; Cell Transplantation; Cells; Chlamydia trachomatis; Clinical; Communicable Diseases; Controlled Study; Cryptococcus neoformans; Development; Disease; Embryo; Engraftment; Evaluation; Founder Generation; Gene-Modified; Genes; Genetic; HIV; HIV Infections; Haplotypes; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hereditary Disease; Human; In Vitro; Infection; Institutes; Institution; Latent Virus; Listeria; Macaca; Macaca fascicularis; Methods; Modeling; Monkeys; Mutation; Mycobacterium tuberculosis; National Heart, Lung, and Blood Institute; National Institute of Allergy and Infectious Disease; National Institute of Child Health and Human Development; National Institute of Neurological Disorders and Stroke; Observational Study; Patients; Plasmodium; Predisposition; Primates; Regimen; Research; Research Personnel; Resource Development; Risk; SIV; Sexual Maturation; Simplexvirus; Stem cells; Techniques; Technology; Toxoplasma gondii; Translations; Transplantation; Trypanosoma cruzi; United States National Institutes of Health; Viral; Viral reservoir; Virus; West Nile viral infection; West Nile virus; Wisconsin; blood group; cancer genetics; genome editing; graft vs host disease; induced pluripotent stem cell; leukemia; mutant; nonhuman primate; novel; public health relevance; purge; simian human immunodeficiency virus; standard of care; vaccine efficacy

 DESCRIPTION (provided by applicant): Hematopoietic stem cell (HSC) transplantation has become a standard of care for the treatment of otherwise incurable blood cancers and genetic diseases. The cure of HIV by transplanting CCR5-mutant (CCR5-delta32) HSCs demonstrated the feasibility and power of stem cell-based therapies for eliminating latent virus and controlling AIDS. However, it will be critical to define the spectrum of anti-viral protection, the engraftment threshold of CCR5-mutant HSCs required for protection, and the potential for depletion of the virus reservoir through "allo-effect" following allogeneic HSC transplantation, to facilitate translation to human patients. We propose to develop an MHC-defined CCR5-mutant nonhuman primate (NHP) bone marrow transplantation model at the Wisconsin National Primate Research Center (WNPRC) to address the critical questions of HSC-based therapies for HIV. This model will employ Mauritian Cynomolgus monkeys (MCMs) with well-defined MHC to control genetic factors in the setting of allogeneic bone marrow transplant. In aim 1, we will use CRISPR/Cas9 methods in MCM induced pluripotent stem cell (iPSC) lines to optimize efficient targeting of the MCM CCR5-locus with minimal off-target effects. In aim 2, we will confirm the feasibility and accuracy of CCR5 genomic editing in IVF-derived MCM embryos and generate CCR5-mutant MCMs. In aim 3, we will evaluate the engraftment of WT and CCR5-mutant HSCs in SHIV-infected, MHC and blood group identical MCM recipients using a myeloablative BMT regimen and assess the effect of these transplants on persistence of SHIV infection. The proposed studies will establish a CCR5-mutant MHC- defined NHP model and will demonstrate utility for evaluation of HSC-based therapies for AIDS. Development of these resources will benefit AIDS researchers at multiple NIH institutions, including ORIP, NHLBI, NIAID, NIDKK, NICHD, and NINDS. Although CCR5-mutant mutation protects from HIV infection, the lack of CCR5 is also associated with increased susceptibility to West Nile virus infection, and other infections including Toxoplasma gondii, Mycobacterium tuberculosis, Chlamydia trachomatis, Listeria, and Plasmodium. Thus, for example, access to CCR5-mutant NHPs will open unique opportunities to establish NHP models for West Nile infection for evaluation of vaccine efficacy and advance the use of NHPs for studies of other infections which may require CCR5-mutant hosts to reproduce a disease course observed in susceptible humans. In addition, advances in gene editing technologies in NHP models will facilitate their clinical translation for treatment of genetic disorders in humans.

 描述（由适用提供）：造血干细胞（HSC）移植已成为治疗原本无法治愈的血液癌和遗传疾病的护理标准。通过移植CCR5突变剂（CCR5-DELTA32）HSC治愈HIV的HSC证明了基于干细胞的疗法消除潜在病毒并控制的可行性和功率 艾滋病。但是，定义抗病毒保护的范围至关重要 CCR5突变HSC的阈值是保护所需的，以及在同种异体HSC移植后通过“异效应”部署病毒储层的潜力，以促进向人类患者的转化。我们建议在威斯康星州国家灵长类动物研究中心（WNPRC）开发一个MHC定义的CCR5-突出非人类灵长类动物（NHP）骨髓移植模型，以解决基于HSC的HSC HIV疗法的关键问题。该模型将在同种异体骨髓移植中使用明确的MHC使用毛里求利亚cynomolgus猴子（MCMS）来控制遗传因素。在AIM 1中，我们将在MCM诱导的多能干细胞（IPSC）线中使用CRISPR/CAS9方法，以优化具有最小脱靶效应的MCM CCR5-locus的有效靶向。在AIM 2中，我们将确认IVF衍生的MCM胚胎中CCR5基因组编辑的可行性和准确性，并生成CCR5突变MCMS。在AIM 3中，我们将使用骨髓性BMT方案评估SHIV感染，MHC和血型相同的MCM受体中WT和CCR5突变的HSC的植入，并评估这些移植对SHIV感染持久性的影响。拟议的研究将建立CCR5突变的MHC定义的NHP模型，并将证明用于评估基于HSC的艾滋病疗法的实用性。这些资源的开发将使多个NIH机构的AIDS研究人员受益，包括Orip，NHLBI，NIAID，NIDKK，NICHD和NINDS。尽管CCR5突变突变可防止HIV感染，但缺乏CCR5也与西尼罗河病毒感染的易感性增加以及其他感染有关，以及其他感染，包括弓形虫弓形虫，结核分枝杆菌，结核病，沙丘乳核酸，沙丘，李斯特氏菌和静脉曲菌。例如，这将访问CCR5-突变的NHP将为西尼罗河感染建立NHP模型，以评估疫苗效率，并推动使用NHP用于研究其他可能需要CCR5突变宿主来再现易感性疾病课程的其他感染的研究。此外，NHP模型中基因编辑技术的进步将促进其临床翻译，以治疗人类的通用疾病。