GENE THERAPY FOR HEMOPHILIA

该项目的目标是为血友病A和b开发基于基因的治疗方法。该项目包括四个项目和两个核心，通过基础研究、转化研究和临床研究来实现总体目标。在项目#1中，D. Katherine High将开展一系列研究，旨在表征AAV转导肌肉细胞长期表达的分子基础，以确定肌肉细胞是否可以适当地翻译后修饰因子IX，并评估对载体中因子IX表达盒的特定修饰是否可以导致更高水平的转基因表达。在项目#2中，Haig Kazazian博士将使用逆转录病毒和腺病毒载体直接进入皮肤和肝脏，以纠正小鼠和犬a型血友病模型中的因子VIII缺乏。初步数据表明，高水平的重组因子VIII可以从皮肤中表达。本项目还将研究一种基于逆转录转座子的新型载体。在项目#3中，Mortimer Poncz博士将探索一种新的血友病基因治疗方法，通过将带有巨核细胞特异性启动子的载体引入造血细胞。Poncz博士的研究将确定以这种方式表达的因子VIII是否能有效治疗a型血友病。在项目#4中，Alan Cohen博士将评估aav介导的肌肉导向基因疗法可以改善B型血友病患者的临床表型而不会产生不良副作用的假设。除了这些项目外，该项目还将包括两个核心，一个是由High博士领导的行政核心，另一个是由Sriram Krishnaswamy博士领导的凝血核心，这两个核心将提供测量小鼠、狗和人类凝血因子抗原和活性水平的专业知识。因子Ix和因子VIII可用的方法。项目和核心之间的交互是广泛的。项目1将开发临床前数据，这些数据将直接用于项目4中正在进行的研究。反过来，项目#4的结果将为项目#1中的效应器提供新的方向。项目1和项目2正在合作开发一种新的载体，并扩展一种成功的基于表皮的因子IX方法。项目#2和#3将从项目#1的互动中受益匪浅，项目#1利用High博士的团队在小型和大型血友病动物模型以及病毒载体介导的基因转移方面的丰富经验。项目#2和3在使用因子VIII敲除小鼠和各种人类和小鼠因子VIII cdna方面进行了强有力的持续合作。一旦载体在动物模型中被证明是有效的，将通过项目#4中获得的经验和可用资源促进转化为临床研究。所有四个项目都将利用行政和凝聚核心。

The objective of this Program Project is to develop gene-based therapies for hemophilias A and B. The program project consists of four projects and two cores that address the overall goal through basic, translational, and clinical studies. In Project #1, D. Katherine High will carry out a series of studies designed to characterize the molecular basis of long-term expression of AAV transduced muscle cells, to determine whether muscle cells can properly post-translationally modify factor IX, and to assess whether specific modifications to the factor IX expression cassette in the vector can lead to higher levels of transgene expression. In Project #2, Dr. Haig Kazazian will use both retroviral and adenoviral vectors directed into skin and liver to correct factor VIII deficiency in murine and canine models of hemophilia A. Preliminary data show that high levels of recombinant factor VIII can be expressed from the skin. This project will also conduct studies on a novel vector based on retro-transposons. In Project #3, Dr. Mortimer Poncz will explore a novel approach to gene therapy for the hemophilias by introducing into hematopoietic cells a vector with a megakaryocyte-specific promoter. Dr. Poncz' studies will determine whether factor VIII expressed in this manner can effectively treat hemophilia A. In Project #4, Dr. Alan Cohen will evaluate the hypothesis that AAV-mediated, muscle-directed gene therapy can ameliorate the clinical phenotype in humans with hemophilia B without untoward side-effects. In addition to these projects, the Program Project will include two cores, an Administrative Core directed by Dr. High and Coagulation Core directed by Dr. Sriram Krishnaswamy, which will provide expertise in measurement of clotting factor antigen and activity levels in mice, dogs and humans. Methods available of factor Ix and factor VIII. Interactions among the projects and cores are extensive. Project #1 will develop pre-clinical data that will be direct utility to the studies ongoing in Project #4. In turn, results from Project #4 will provide new directions for the effectors in Project #1. Projects #1 and 2 are collaborating for development of a novel vector and on extension of a successful epidermal-based approach to factor IX. Projects #2 and #3 will benefit greatly from interactions with Project #1 utilizing the considerable experience of Dr. High's group in small and large animal models of hemophilia, and with viral vector-mediated gene transfer. Projects #2 and 3 have strong ongoing collaborations centered on the use of the factor VIII knockout mice and various human and mouse factor VIII cDNAs. Once vectors have been demonstrated to be efficacious in animal models,, translation to clinical investigation will be facilitated by experience gained and resources available in Project #4. All four projects will make use of the Administrative and the Coagulation Core.