GAMMA GLOBIN VECTORS FOR TREATMENT OF HEMOGLOBINOPATHIES

这项申请的重点是应聘者的直接职业目标，即通过获得开发、测试和使用珠蛋白载体的新技能，加强和进一步他迄今以实验室为基础的培训，这些载体是为β-链状血红蛋白疾病的基因治疗方法而设计的。凭借申请者的临床背景、以前的博士研究经验以及最近在圣犹大儿童研究医院(SJCRH)Arthur Nienhuis博士的实验室进行的三年博士后工作，候选人现在进入了他职业生涯的过渡阶段，目标是成为一名独立的研究员，成为一名临床医生兼科学家。然而，候选人和赞助商坚信，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，并极大地提高作为独立研究人员早期成功的潜力。作为一名学术医学领域的独立教员，候选人的长期职业目标是继续在血液疾病的基因治疗领域，特别是开发与成功的临床前基因治疗方法转化为临床的研究计划相兼容的研究计划。在这份申请中，候选人提议与他目前在SJCRH的导师Arthur Nienhuis博士一起获得开发和测试新的治疗性珠蛋白载体方面的额外培训和特定专业知识。在分部内。作为实验血液学研究所的成员和主任，他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以及人类干细胞NOD/SCID小鼠移植模型的应用方面拥有丰富的专业知识。因此，申请人执行拟议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进的珠蛋白载体的需要，用于地中海贫血和镰状细胞性贫血的基因治疗方法。这个项目的重点是一种基因添加策略，该策略基于这样一种假设，即交付优化的伽马珠蛋白基因盒可以在发育中的红系细胞中实现足够的表达水平，以逆转地中海贫血或镰状细胞疾病的表型。该项目包括3个具体目标：1)设计和测试新型伽马珠蛋白逆转录病毒和慢病毒载体，2)使用β-地中海贫血小鼠模型来模拟使用优化的伽马珠蛋白载体的基因治疗方法。3)鉴定和使用来自β-地中海贫血患者的原始造血细胞，以评估优化的伽玛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.