LENTIVIRAL GENE TRANSFER INTO PANCREATIC BETA-CELLS

描述（摘自申请人摘要） 糖尿病是一种困扰着超过200万美国人的疾病。 一种治疗这种疾病的基因治疗方法需要：1）安全的 将候选基因导入胰岛细胞以防止其 排斥，和2）用于产生大量胰岛细胞供应的方法。 本项目满足了这两项要求。通过工程化 慢病毒系统，其由提供人类免疫应答的细胞系组成。 免疫缺陷病毒1型（HIV-1）的包装功能，一个人 免疫缺陷病毒2型（HIV-2）转移载体和包膜 表达下述糖蛋白B（gB）包膜蛋白的载体 巨细胞病毒（CMV），我们假设一个将产生病毒粒子， 以高亲和力结合β-胰岛细胞， 转基因的表达而没有炎症反应。的 将可切除的癌基因引入胰岛细胞可以允许 他们的传播与能力，以消除转化片段的 DNA. 所描述的工作将在以下出色的环境中进行： 哈佛医学院Vikas P. Sukhatme，M.D.实验室，哲学博士、 致力于基因治疗应用的分子生物学家。合作者 包括特里B。斯特罗姆，医学博士，使用免疫构建体的专家， 细胞免疫和Towia A.利伯曼博士，他有相当多的 在同种异体胰岛细胞移植中的操作经验。 顾问Norman Letvin，医学博士，将提供慢病毒方面的专业知识 基因组和嵌合体的构建。新方法，申请人将 学习是细胞系开发、重组腺病毒的生产， 胰岛的分离和移植。 申请人是一个联合的M.D./博士程序 位于达拉斯的德克萨斯大学西南医学中心， 在哺乳动物细胞蛋白质折叠方面的经验。申请人已花费 他在肾脏病学研究金的两年里开发了慢病毒载体系统 并研究细胞因子的释放机制， 腺病毒感染他绝对致力于一个以 肾脏病学学术部门的基础研究。 研究中的发起人、合作者和顾问的综合素质 哈佛医学院沿着的环境以及申请人以前的 研究经验和对研究的持续承诺提供了一个独特的 申请人有机会实现项目的目标，并成为 成功的独立调查员

DESCRIPTION (Taken from the applicant's Abstract) Diabetes mellitus is a disease that afflicts over two million Americans. One gene therapy approach to treating this disease requires: 1) a safe vector to introduce candidate genes into islet cells to prevent their rejection, and 2) a method for producing an abundant supply of islet cells. This project addresses both of these requirements. By engineering a lentiviral system that consists of a cell line providing human immunodeficiency virus type 1 (HIV-1) packaging functions, a human immunodeficiency virus type 2 (HIV-2) transfer vector, and an envelope vector expressing the glycoprotein B (gB) envelope protein of cytomegalovirus (CMV), we hypothesize that one will produce virions that bind with high affinity to beta-islet cells and provide long-lasting expression of a transgene without an inflammatory response. The introduction of an excisable oncogene into pancreatic islet cells may allow their propagation with the ability to remove the transforming fragment of DNA. The work described will be performed in the outstanding environment of Harvard Medical School in the laboratory of Vikas P. Sukhatme, M.D., Ph.D., a molecular biologist committed to gene therapy applications. Collaborators include Terry B. Strom, M.D., expert in the use of immunoconstructs leading to cellular immunity and Towia A. Libermann, Ph.D., who has considerable experience in the manipulation of islet cells for allotransplantation. Consultant Norman Letvin, M.D., will provide expertise in the lentiviral genome and construction of chimeras. New methodologies the applicant will learn are cell line development, production of recombinant adenovirus, and isolation and transplantation of pancreatic islets. The applicant is a graduate of a combined M.D./Ph.D. program of the University of Texas Southwestern Medical Center at Dallas with 3.5 years of experience in protein folding in mammalian cells. The applicant has spent two years of his Nephrology fellowship developing a lentiviral vector system and investigating the mechanism of release of cytokines in response to adenoviral infection. He is absolutely committed to a career centered on basic research in an academic division of Nephrology. The combined quality of the sponsor, collaborators, and consultant in the environment of Harvard Medical School along with the applicant's previous research experience and ongoing commitment to research provides a unique opportunity for the applicant to achieve the goals of the project and become a successful independent investigator.