Engineering an HIV-resistant immune system for HIV cure

设计抗艾滋病毒免疫系统以治愈艾滋病毒

• 批准号: 9063293
• 负责人: Hind Jawdat Fadel
• 金额: $ 17.04万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9063293
• 关键词: AIDS/HIV problem; APOCEC3G gene; Adherence; Aging; Alleles; Allogeneic Bone Marrow Transplantation; Amino Acids; Animals; Anti-Retroviral Agents; Antiviral Agents; Area; Autoimmune Diseases; Award; Berlin; Binding; Biological Assay; Biology; Bone Marrow Transplantation; CCR5 gene; CD34 gene; CD4 Positive T Lymphocytes; Capsid; Cardiovascular system; Cell Line; Cells; Chromosomes; Chromosomes, Human, Pair 5; Chronic; Clinic; Colorado; Communicable Diseases; Complex; Coupled; DNA; Data; Dependency; Development; Development Plans; Disease; Docking; Doctor of Medicine; Doctor of Philosophy; Drug toxicity; Engineering; Environment; Epidemic; Evolution; Gene Targeting; Genes; Genome; Genome engineering; Goals; HIV; HIV resistance; HIV-1; Heel; Hematological Disease; Hematopoietic stem cells; Hepatic; Human; Human Genome; Immune System Diseases; Immune system; Infection; Inflammation; Ingestion; Integration Host Factors; Kidney; Knock-in; Knock-out; Lead; Left; Life; Macaca mulatta; Malignant Neoplasms; Mediating; Medicine; Mentors; Metabolic; Methods; Modification; Molecular Medicine; Natural regeneration; Neurologic; Pathogenesis; Patients; Pharmaceutical Preparations; Pharmacotherapy; Physicians; Point Mutation; Positioning Attribute; Problem Solving; Process; Proteins; Recruitment Activity; Research; Research Personnel; Resistance; Role; Scientist; Site; Specificity; System; Technology; Testing; Therapeutic; Training; Universities; Virus; Virus Replication; Work; antiretroviral therapy; career; career development; clinical application; cofactor; collaborative environment; college; conditioning; experience; gene therapy; genome editing; humanized mouse; immunosenescence; in vivo; insight; leukemia; medical schools; member; novel; nuclease; pandemic disease; particle; pill; pre-clinical; prevent; professor; public health relevance; reconstitution; research and development; resistance gene; skills; success; theories; therapeutic target; therapy development; transcription activator-like effector nucleases; transcriptional coactivator p75; transmission process; viral resistance

 DESCRIPTION (provided by applicant): The candidate, Hind Fadel, M.D., Ph.D., is an Assistant Professor at the Mayo Clinic College of Medicine. She is a member of the Division of Infectious Diseases in the Department of Medicine, and of the Department of Molecular Medicine at the Mayo Clinic. Dr. Fadel is a physician-scientist committed to a career in basic and translational HIV-1 research and gene therapy. She has gathered an experienced team of mentors, collaborators and advisors and is proposing a comprehensive research, course work, and career development plan that will position her to be a successful independent investigator. The environment provided by the Mayo Clinic is ideally suited for the success of both the research and career development components of the award. She has also maintained and identified advisors and collaborators outside Mayo Clinic. The career development plan will allow her to hone her skills in genome editing with engineered site-specific nucleases, to gain insights into host factors role in HIV-1 biology, and to develop new skills to manipulate human genome for HIV-1 cure applications and gene therapy. The research plan focuses on developing HIV cure strategies that use targeting/editing of HIV-1 dependency and restriction factor genes with novel engineered nucleases in order to create HIV-1 resistance. The methods will be applied to primary CD34+ hematopoietic stem cells (HSCs) and studied in humanized mice. While antiretroviral drugs have made treatment of HIV disease possible, they in many ways represent a "halfway technology." They leave existing patients vulnerable to major problems that include resistance, persistent immune dysfunction and immunosenescence, complex metabolic disturbances, and accelerated aging phenomena. The second problem is that while drug treatment can in theory prevent further transmission, the many problems associated with supplying the pill combinations life-long and with maintaining adherence have meant that the epidemic continues to expand. A definitive cure could solve both problems. Cure was until recently considered futuristic, but this has now changed. The recent cure of a patient with both HIV and leukemia -- the so-called "Berlin patient" -- by bone marrow transplantation from a donor lacking a main HIV cellular cofactor (the entry co-receptor CCR5) was energizing to the field. The specific approach (allogeneic BMT) is far too toxic for anyone without cancer. However, the value of targeting a gene that HIV needs (a host cofactor) has been made clear. Along with the very recent emergence of highly promising gene targeting technologies, and advances in our understanding of the disease process itself, the Berlin patient helped catalyze making cure a central priority in the field. Modification of a patient's own HSCs to regenerate an HIV- resistant immune system in subjects with HIV/AIDS, coupled with appropriately mild conditioning, could be an effective cure strategy. It is essential to recruit and develop young investigators to populate what is certain to be an important, expanding field. Dr. Fadel is proposing to target two vulnerabilities of HIV-1 as she develops curative gene therapy approaches: (i) eliminating host dependency factors HIV-1 requires, and (ii) editing host restriction factors that HIV-1 has evolved to evade in order to restore their ability to restrict HV-1. Her central hypothesis is that engineered nucleases will efficiently target HIV-1 dependency and restriction factors to generate resistance to HIV-1 infection for therapeutic application and t further our understanding of these factors in HIV-1 pathogenesis. The main aims are: 1) Knockout HIV-1 dependency factors in human CD4+ T-cell lines to simultaneously determine their virological roles and establish their potential for therapeutic targets for HIV cure applications; 2) Edit human restriction factors to restore HIV-1 resistance and determine effects on viral replication; 3) Create dependency factor knockouts and restriction factor knock-ins in CD34+ HSC cells ex-vivo and characterize them in pre-clinical animal studies in humanized mice. The proposed research has the potential to provide insights into the roles of multiple host factors in HIV-1 biology and to lead to translational applications for curative HIV-1 therapies. Developing HSC-mediated therapeutics in this disease can also advance related applications to other infectious, autoimmune and hematologic diseases.

 描述（由适用提供）：候选人，医学博士Hind Fadel，博士，是Mayo诊所医学院的助理教授。她是医学系传染病科和梅奥诊所分子医学系的成员。 Fadel博士是一位身体科学家，致力于从事基础和翻译的HIV-1研究和基因疗法的职业。她聚集了一支经验丰富的导师，合作者和顾问团队，并正在提出一项全面的研究，课程工作和职业发展计划，这将使她成为成功的独立研究员。 Mayo诊所提供的环境非常适合该奖项的研究和职业发展组成部分的成功。她还维持并确定了Mayo诊所以外的顾问和合作者。职业发展计划将使她能够使用工程特定地点的核武器来尊重自己在基因组编辑方面的技能，从而深入了解HIV-1生物学中宿主因素的作用，并开发新技能，以操纵HIV-1治疗应用和基因疗法的人类基因组。该研究计划着重于制定HIV治疗策略，该策略使用新型工程核心使用HIV-1依赖性和限制因素基因的靶向/编辑，以创建HIV-1耐药性。该方法将应用于原发性CD34+造血干细胞（HSC），并在人源化的小鼠中进行了研究。尽管抗逆转录病毒药物使HIV疾病的治疗成为可能，但它们在许多方面都代表了“中途技术”。他们使现有患者容易受到主要问题的影响，包括耐药性，持续性免疫功能障碍和免疫衰老，复杂的代谢疾病以及加速的衰老现象。第二个问题是，尽管在理论上可以防止进一步传播药物，但与供应药丸组合终生和维持依从性相关的许多问题意味着流行病在不断扩大。定义的治疗可以解决这两个问题。直到最近被认为是未来派的，但现在已经改变了。艾滋病毒和白血病患者最近通过缺乏缺乏HIV HIV细胞辅助因子的供体的骨髓移植（入口共受感受器CCR5）来治愈了所谓的“柏林患者”。对于没有癌症的任何人来说，具体方法（同种异体BMT）太毒性了。但是，靶向艾滋病毒需要的基因（宿主辅助因子）的价值已清楚。除了最近出现的高度有希望的基因靶向技术以及我们对疾病过程本身的理解的进步之外，柏林患者还帮助催化使治疗成为该领域的核心优先事项。修改患者自己的HSC，以在患有艾滋病毒/艾滋病的受试者中再生抗艾滋病毒的免疫系统，再加上适当的轻度调节，可能是一种有效的治疗策略。招募和培养年轻的研究人员以填充肯定是一个重要的，不断扩展的领域是至关重要的。 Fadel博士提议针对HIV-1的两个漏洞，因为她开发了治愈性基因疗法方法：（i）消除HIV-1需要的宿主依赖性因素，以及（ii）编辑HIV-1已演变为逃避以恢复其限制HV-1的宿主限制因素。她的中心假设是，工程核武器将有效地靶向HIV-1的依赖性和限制因素，以产生对HIV-1感染的抗药性，并进一步了解HIV-1发病机理中这些因素。主要目的是：1）人类CD4+ T细胞系中的基因敲除HIV-1依赖性因子，以同时确定其病毒学作用并确定其用于HIV治疗应用的治疗靶标的潜力； 2）编辑人类限制因素，以恢复HIV-1抗性并确定对病毒复制的影响； 3）在CD34+ HSC细胞中创建依赖性因子基因敲除和限制因子敲除ins，并在人道化小鼠的临床前动物研究中表征它们。拟议的研究有可能洞悉HIV-1生物学中多种宿主因素的作用，并导致治疗HIV-1疗法的转化应用。在这种疾病中开发HSC介导的疗法也可以推动针对其他感染性，自身免疫和血液学疾病的相关应用。