PARVOVIRUS VECTORS FOR HUMAN GENE THERAPY

描述：腺相关病毒2（AAV），一种非致病性人类 细小病毒已被建议作为人类基因的潜在有用载体 疗法 然而，与AAV宿主相关的一些基本问题 细胞间的相互作用在很大程度上仍未被探索。 其中包括身份 AAV的细胞受体，以及AAV与 正常人二倍体细胞，天然AAV最可能的靶细胞， 感染，也是利用该载体进行基因治疗的潜在靶点 系统 类似地，虽然第二种人源细小病毒，命名为 细小病毒B19对人类具有明显的嗜性 红系造血祖细胞B19载体 能够红系细胞特异性递送基因的细胞还没有被 开发 Srivastava博士和他的同事们提出， 细胞受体的AAV，并评估病毒宿主相互作用，使用 人脐带血原代造血干/祖细胞 脐带血 细小病毒B19的嗜红细胞性有待进一步研究 开发基于B19的新型载体，以实现靶向递送人 珠蛋白基因 本研究拟检验的假设为：1. 的 AAV进入人细胞是特异性的和受体介导的。 2. 的 红系细胞特异性递送和高水平表达转导的 在造血细胞中的β-珠蛋白基因是可行的，在体外以及在 vivo. 四个具体目标是：1。 相互作用的表征 重组AAV载体与造血干细胞和祖细胞在 人脐带血，和鉴定推定的细胞 AAV的受体。 2. AAV介导的转导和红系细胞的评价 正常人β-珠蛋白基因的细胞特异性高水平表达， 脐带血造血祖细胞 3. 细小病毒的研究进展 基于B19的载体和B19介导的红系细胞靶向递送以及 正常人β-珠蛋白基因在造血祖细胞中的表达 脐带血中的细胞 4. 研究AAV和B19介导的 体外转导，和长期体内表达的潜力， 在β-地中海贫血小鼠中转导的人β-珠蛋白基因， 非人类灵长类动物模型。 这些研究将提供新的见解的基础分子生物学， 细小病毒-二倍体细胞相互作用作为开发安全 和有效载体，并有助于评估体内功效， 在其用于人类基因治疗的潜在用途之前的安全性 一般血红蛋白病、镰状细胞病和β-地中海贫血 特别是。

DESCRIPTION: The adeno-associated virus 2 (AAV), a non-pathogenic human parvovirus, has been suggested as a potentially useful vector for human gene therapy. However, a number of fundamental questions related to AAV-host cell interactions remain largely unexplored. These include the identity of the cellular receptor for AAV, and the nature of interaction of AAV with normal human diploid cells, the most likely target for a natural AAV infection, and also a potential target for gene therapy with this vector system. Similarly, although a second parvovirus of human origin, designated parvovirus B19, has been shown to possess a remarkable tropism for human hematopoietic progenitor cell in the erythroid lineage, B19-based vectors capable of erythroid cell-specific delivery of genes have not been developed. Dr. Srivastava and his colleagues propose to identify the cellular receptor for AAV, and evaluate the virus host interaction using primary human hematopoietic stem and progenitor cells in human umbilical cord blood. The erythroid cell-tropism of parvovirus B19 will be exploited to develop novel B19-based vectors to achieve targeted delivery of human globin genes. The hypotheses to be tested in this proposal are: 1. That entry of AAV in human cells is specific and receptor-mediated. 2. That erythroid cell-specific delivery and high level expression of a transduced beta-globin gene in hematopoietic cells is feasible in vitro as well as in vivo. The four specific aims are: 1. Characterization of interaction of recombinant AAV vectors with hematopoietic stem and progenitor cells in human umbilical cord blood, and identification of the putative cellular receptor for AAV. 2. Evaluation of AAV-mediated transduction and erythroid cell-specific, high level expression of a normal human beta-globin gene in hematopoietic progenitor cells in cord blood. 3. Development of parvovirus B19-based vector and B19-mediated erythroid cell-targeted delivery and expression of a normal human beta-globin gene in hematopoietic progenitor cells in cord blood. 4. Investigation of AAV- and B19- mediated transduction ex vivo, and the potential for long-term in vivo expression of the transduced human beta-globin gene in beta-thalassemic murine and non-human primate models. These studies will provide new insights into the basic molecular biology of parvovirus-diploid cell interactions as a prelude to the development of safe and effective vectors, and help in evaluating the in vivo efficacy and safety prior to their potential use in gene therapy for human hemoglobinopathies in general, and sickle cell disease and beta-thalassemia in particular.