Targeting Entry in Epithelia with ICMV-FIV

使用 ICMV-FIV 靶向进入上皮细胞

• 批准号: 6853152
• 负责人: PAUL B MCCRAY
• 金额: $ 19.24万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6853152
• 关键词: biotechnology; cell differentiation; chloride channels; clinical research; cystic fibrosis; feline immunodeficiency virus; gene delivery system; gene expression; genetic transduction; glycoproteins; human tissue; laboratory mouse; lymphocytic choriomeningitis virus; receptor binding; respiratory epithelium; transfection /expression vector; virus envelope; virus infection mechanism; virus protein; virus receptors

Gene transfer to pulmonary epithelia is a direct approach to correct the ion transport and host defense defects associated with cystic fibrosis (CF). Unfortunately, several problems currently preclude its clinical application including poor apical gene transfer efficiency, transient expression, adverse immune responses, and a limited understanding of the intracellular fate of integrating vectors. Our recent work funded through this program focused on the development and application of recombinant lentiviral vectors based on feline immunodeficiency virus (FIV) to surmount these problems. Initial steps in retroviral transduction require interactions between the viral envelope glycoprotein (GP) and receptors on the host cell. For many vector envelopes, the cellular receptors are functionally absent from the apical surface of airway epithelia. Significant progress has come through our detailed studies of the interactions between the airway cell and envelope GPs. Recently we successfully pseudotyped FIV using the GPs from several enveloped viruses to direct vector binding and entry to the apical surface of airway epithelia. Importantly, FIV vector pseudotyped with envelope glycoproteins from the arenavirus lymphocytic choriomeningitis virus (LCMV) strain WE54 enters polarized human airway epithelia from the apical surface without the disruption of tight junctions. Therefore we now have a useful reagent to address important issues regarding the interactions between an integrating vector and airway epithelia. Our central hypothesis is that LCMV pseudotyped FIV can enter airway epithelia via the apical surface, allowing for persistent correction of the CF defect by gene transfer. We will focus on four main questions: 1) What is the receptor for LCMV pseudotyped FIV in airway epithelia? 2) Are there apical barriers to gene transfer with LCMV pseudotyped FIV in airway epithelia? 3) Does LCMV pseudotyped FIV confer persistent transgene expression in human airway epithelia? 4) If FIV enters airway epithelia through the apical surface in vivo will it result in persistent transgene expression? Completing these aims will address key questions regarding FIV-airway cell interactions and improve our ability to apply lentiviral vectors for the treatment and prevention of cystic fibrosis lung disease.

基因转移到肺上皮是纠正与囊性纤维化（CF）相关的离子转运和宿主防御缺陷的直接方法。不幸的是，目前有几个问题排除了其临床应用，包括不良的顶部基因转移效率，瞬态表达，不良免疫反应以及对整合向量的细胞内命运的有限理解。我们最近通过该计划资助的工作重点是基于猫免疫缺陷病毒（FIV）的重组慢病毒载体的开发和应用，以克服这些问题。逆转录转导的初始步骤需要在 病毒包膜糖蛋白（GP）和宿主细胞上的受体。对于许多载体信封，在气道上皮的顶部表面缺乏细胞受体。通过我们对气道电池与包膜GPS之间相互作用的详细研究，取得了重大进展。最近，我们使用来自几种包膜病毒的GPS成功地拟型FIV，以指导载体结合并进入气道上皮的顶部表面。重要的是，从体育症病毒病毒淋巴病毒脉络膜脑膜炎病毒（LCMV）菌株WE54进入极化的人类气道上皮的FIV载体伪型 从顶部表面，而不会破坏紧密连接。因此，我们现在有一个有用的试剂来解决有关综合向量和气道上皮相互作用的重要问题。我们的中心假设是，LCMV伪型FIV可以通过顶部表面进入气道上皮，从而可以通过基因转移对CF缺陷进行持续校正。我们将重点关注四个主要问题：1）气道上皮中LCMV伪型FIV的受体是什么？ 2）在气道上皮中有LCMV伪型FIV存在基因转移的根尖障碍吗？ 3）LCMV是否在人气道上皮中允许持续的转基因表达？ 4）如果FIV通过体内顶端表面进入气道上皮，是否会导致持续的转基因表达？完成这些目标将解决有关FIV-Airway细胞相互作用的关键问题，并提高我们将慢病毒载体应用于治疗和预防囊性纤维化肺疾病的能力。