LENTIVIRAL GENE TRANSFER INTO PANCREATIC BETA-CELLS

描述(摘自申请人的摘要) 糖尿病是一种折磨着200多万美国人的疾病。 治疗这种疾病的一种基因疗法需要：1)安全的 载体将候选基因导入胰岛细胞以防止其 排斥反应，以及2)一种产生充足的胰岛细胞供应的方法。 该项目满足了这两个要求。通过设计一种 慢病毒系统，由一个细胞系组成，为人类提供 免疫缺陷病毒1型(HIV-1)包装功能，人类 一种人类免疫缺陷病毒2型转移载体和包膜 猪瘟病毒糖蛋白B(GB)包膜蛋白的表达载体 巨细胞病毒(CMV)，我们假设一个人会产生病毒粒子 与β-胰岛细胞高亲和力结合，并提供持久的 在没有炎症反应的情况下表达转基因。这个 将可切除的癌基因导入胰岛细胞可能会 它们的传播具有移除转换片段的能力 DNA 所描述的工作将在以下环境中进行 哈佛医学院维卡斯·P·苏哈特姆，医学博士，博士， 一位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，免疫构造使用方面的专家 细胞免疫和托维亚·A·利伯曼博士，她有相当多的 同种异体胰岛细胞移植的操作体会。 医学顾问诺曼·莱特文将提供慢病毒方面的专业知识 基因组和嵌合体的构建。申请者将采用新的方法 学习是细胞系的发展，重组腺病毒的生产，以及 胰岛分离与移植。 申请人毕业于哈佛大学医学博士/博士结合项目。 德克萨斯大学达拉斯西南医学中心，拥有3.5年的 在哺乳动物细胞中进行蛋白质折叠的经验。申请人已花费 他两年的肾脏病研究开发慢病毒载体系统 并研究了细胞因子释放的机制 腺病毒感染。他绝对致力于他的职业生涯 肾脏病学部的基础研究。 赞助商、合作者和顾问的综合素质 哈佛医学院的环境和申请人以前的环境 研究经验和对研究的持续承诺提供了独特的 申请者有机会实现项目目标并成为 一位成功的独立调查员。

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.