Selection of RNAi-based inhibitors for anti-HIV gene therapy

作     者：Stefanie A Knoepfel Mireille Centlivre Ying Poi Liu Fatima Boutimah Ben Berkhout 

作者机构：Laboratory of Experimental Virology Department of Medical Microbiology Center for Infection and Immunity Amsterdam Academic Medical Center University of Amsterdam 1105 AZ Amsterdam The Netherlands 

出 版 物：《World Journal of Virology》 (世界病毒学杂志)

年 卷 期：2012年第1卷第3期

页      面：79-90页

学科分类：10[医学] 

基　　金：Supported by The NWO-CW(Chemical Sciences),Zon Mw(Medical Sciences),the Dutch AIDS Fund(project 2006006) the DAAD(German Academic Exchange Service) the FRM(Fondation pour la Recherche Medicale) 

主　　题：Human immunodeficiency virus type 1 RNA interference Gene therapy “Human Immune System”mouse Lentivirus 

摘      要：In the last decade, RNA interference(RNAi) advanced to one of the most widely applied techniques in the biomedical research field and several RNAi therapeutic clinical trials have been launched. We focus on RNAibased inhibitors against the chronic infection with human immunodeficiency virus type 1(HIV-1). A lentiviral gene therapy is proposed for HIV-infected patients that will protect and reconstitute the vital immune cell pool. The RNAi-based inhibitors that have been developed are short hairpin RNA molecules(sh RNAs), of which multiple are needed to prevent viral escape. In ten distinct steps, we describe the selection process that started with 135 sh RNA candidates, from the initial design criteria, via testing of the in vitro and in vivo antiviral activity and cytotoxicity to the final design of a combinatorial therapy with three sh RNAs. These sh RNAs satisfied all 10 selection criteria such as targeting conserved regions of the HIV-1 RNA genome,exhibiting robust inhibition of HIV-1 replication and having no impact on cell physiology. This combinatorial sh RNA vector will soon move forward to the first clinical studies.