GAMMA GLOBIN VECTORS FOR TREATMENT OF HEMOGLOBINOPATHIES

该申请的重点是候选人的直接职业目标，即通过获得开发、测试和使用球蛋白载体的新技能来增强和进一步他的实验室培训，这些球蛋白载体是为基因治疗方法设计的，用于治疗β链血红蛋白病。凭借申请人的临床背景，之前的博士研究经验，以及最近在St. Jude儿童研究医院（SJCRH） Arthur Nienhuis博士实验室的三年博士后工作，候选人目前正进入其职业生涯的过渡阶段，目标是成为一名临床科学家的独立研究者。然而，候选人和发起人坚信，涉及本申请中概述的新载体和动物模型的进一步培训将促进这一过渡，并大大提高作为独立研究者早期成功的潜力。作为学术医学环境中的独立教员，候选人的长期职业目标是继续在血液病基因治疗领域进行研究，并对开发与成功的临床前基因治疗方法转化相匹配的研究项目感兴趣。在此申请中，候选人建议在SJCRH的现任导师Arthur Nienhuis博士的指导下，获得新的治疗性珠蛋白载体开发和测试方面的额外培训和特定专业知识。Nienhuis博士是实验血液学部门的成员和主管，在逆转录病毒和慢病毒载体开发、小鼠和人类造血干细胞基因转移技术、地中海贫血动物模型和NOD/SCID人类干细胞小鼠移植模型的使用方面具有重要的专业知识。因此，申请人为执行拟议的研究所需的进一步培训是现成的。拟议的研究项目是基于开发改良珠蛋白载体的需求，用于地中海贫血和镰状细胞性贫血的基因治疗方法。该项目的重点涉及一种基因添加策略，该策略基于一种假设，即递送优化的γ -珠蛋白基因盒可以在发育中的红细胞中达到足够的表达水平，从而逆转地中海贫血或镰状细胞病表型。该项目包含3个具体目标：1)设计和测试新的γ -珠蛋白逆转录病毒和慢病毒载体，2)使用小鼠模型来模拟使用优化的γ -珠蛋白载体的基因治疗方法。3)表征和利用β -地中海贫血患者的原始造血细胞，评价优化后的γ -珠蛋白载体的治疗潜力。

This application is focused on the candidate's immediate career goal, which is to enhance and further his laboratory-based training to date by acquiring new skills in the development, testing and use of globin vectors designed for gene therapy approaches to the beta-chain hemoglobinopathies. With the applicant's clinical background, previous doctoral research experience and three years of post-doctoral work in the laboratory of Dr. Arthur Nienhuis at St. Jude Children's Research Hospital (SJCRH) most recently, the candidate is now entering a transitional phase in his career with the goal of becoming an independent investigator as a clinician-scientist. However, the candidate and the sponsor strongly believe that further training involving the new vectors and animal models outlined in this application will facilitate this transition and greatly enhance the potential for early success as an independent investigator. As an independent faculty member in an academic medical setting, it is the candidate's long-term career goal to continue in the area of gene therapy for hematologic disorders with specific interest in developing a research program compatible with the translation of successful preclinical gene therapy approaches to the clinic. In this application, the candidate proposes to obtain additional training and specific expertise in the development and testing of new therapeutic globin vectors with his current mentor, Dr. Arthur Nienhuis, at SJCRH. Within the Div. of Experimental Hematology in which Dr. Nienhuis is a member and Chief, there is significant expertise in retroviral and lentiviral vector development, in techniques of gene transfer into murine and human hematopoietic stem cells, in animal models of thalassemia, and in the use of the NOD/SCID murine transplant model for human stem cells. Thus, the further training the applicant requires for the execution of the proposed research is readily available. The proposed research project is based on the need for the development of improved globin vectors for use in a gene therapy approach to both thalassemia and sickle cell anemia. The focus of this project involves a gene addition strategy based on the hypothesis that delivery of an optimized gamma-globin gene cassette can achieve a sufficient level of expression in developing erythroid cells to reverse the thalassemic or sickle cell disease phenotype. The project contains 3 specific aims: 1) to design and test novel gamma-globin retroviral and lentiviral vectors, 2) to use a murine model of beta-thalassemia to model gene therapy approaches using optimized gamma-globin vectors. and 3) to characterize and use primitive hematopoietic cells from patients with beta-thalassemia to evaluate the therapeutic potential of optimized gamma-globin vectors.