Bacteria Based Agents for Therapy of AIDS Associated Infections

用于治疗艾滋病相关感染的细菌制剂

基本信息

批准号：
8141140
负责人：
SANGWEI LU
金额：
$ 8.39万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-03 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8141140
关键词：
AIDS therapy Acquired Immunodeficiency Syndrome Address Animal Model Animals Antiviral Agents Attenuated Bacteria Biological Models Catalytic RNA Cells Cultured Cells Cytomegalovirus Cytomegalovirus Infections Development Drug resistance Essential Genes Ganciclovir Gastroenteritis Gene Delivery Gene Expression Gene Targeting Gene Transfer Growth Homebound Persons Human Human Virus Immune Immunocompromised Host Infection Inflammatory Response Mammalian Cell Mediating Methods Modeling Murid herpesvirus 1 Mus Nucleic Acids Opportunistic Infections Oral Organ Pathogenesis Patients Pharmaceutical Preparations Plasmids Pneumonia Prevention RNase P Reporting Research Research Proposals Retinitis Salmonella Satellite Viruses Small Interfering RNA System Therapeutic Therapeutic Agents Vaccines Viral Genes Viral Pathogenesis Virus Diseases Vision anti-viral gene delivery base clinical application combat cytotoxicity expression vector gene transfer vector human disease in vivo insight mouse model novel novel strategies novel therapeutic intervention pathogen targeted delivery tool tumor

项目摘要

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) causes one of the most common opportunistic infections in patients with AIDS. Disseminated HCMV infection in these patients is usually associated with gastroenteritis, pneumonia, and sight-threatening retinitis. The emergence of drug-resistant HCMV strains to currently available drugs (e.g. ganciclovir) has posed a need to develop new drugs and novel strategies to combat HCMV infections. The objective of this study is to develop Salmonella as a novel class of gene transfer vectors for targeted delivery of anti-HCMV RNase P ribozymes for blocking viral infection. Attenuated strains of Salmonella can function as a carrier system for the delivery of eukaryotic expression vectors and have been shown to deliver therapeutic agents, including nucleic acids-based vaccines and anti-tumor molecules (e.g. small interfering RNAs) for the treatment and prevention of human diseases. We have recently shown that attenuated Salmonella efficiently delivered an anti-HCMV RNase P ribozyme sequence to human cells, leading to substantial ribozyme expression and effective inhibition of viral infection. Furthermore, we have generated novel attenuated Salmonella strains that efficiently delivered RNase P ribozyme sequence for expression in cells. However, little is currently known about the mechanism of how Salmonella achieves efficient gene transfer for delivery of nucleic acids-based agents (e.g. RNase P ribozymes) in human cells. It has not been reported if Salmonella-mediated delivery of RNase P ribozymes is specific and effective in inhibiting viral infection and pathogenesis in animal models. To address these issues, we propose to first study how the generated Salmonella strains achieve efficient gene transfer. We will then investigate whether targeted delivery of RNase P ribozyme by the generated Salmonella abolishes HCMV gene expression and growth in cultured human cells. Using murine cytomegalovirus (MCMV) infection of mice as a model system, we will determine whether the generated Salmonella strains are highly efficient for targeted delivery of RNase P ribozyme in animals and whether the targeted delivery of RNase P ribozymes mediated by Salmonella is highly effective in blocking MCMV infection and pathogenesis in vivo. The potential immune/inflammatory responses and cytotoxicity associated with the generated Salmonella strains will also be investigated. Our research will generate novel Salmonella strains with efficient gene transfer activity that can be used in gene-targeting clinical applications. Furthermore, these results will provide insight into the mechanism of how Salmonella achieves efficient gene delivery in mammalian cells, and will determine whether Salmonella-mediated delivery of RNase P ribozymes is specific and effective in shutting down gene expression in cultured cells and in animals. This study will facilitate the development of Salmonella-mediated delivery of RNase P ribozymes as a novel therapeutic approach for treatment of infections by HCMV and other AIDS-associated viruses. PUBLIC HEALTH RELEVANCE: The proposed research is to generate novel agents for targeted gene delivery of antiviral molecules in treating infections caused by AIDS-associated viruses, including human cytomegalovirus (HCMV), which causes one of the most common opportunistic infections in AIDS patients. Our study will facilitate the development of a new method that can be used as a research tool and a therapeutic approach for studies and treatment of infections associated with human viruses such as HCMV.

描述（由申请人提供）：人类巨细胞病毒（HCMV）引起艾滋病患者最常见的机会性感染之一。这些患者的播散性 HCMV 感染通常与胃肠炎、肺炎和危及视力的视网膜炎有关。对现有药物（例如更昔洛韦）产生耐药性的 HCMV 菌株的出现，需要开发新药和新策略来对抗 HCMV 感染。本研究的目的是将沙门氏菌开发为一类新型基因转移载体，用于靶向递送抗 HCMV RNase P 核酶以阻断病毒感染。沙门氏菌减毒株可以作为递送真核表达载体的载体系统，并已被证明可以递送治疗剂，包括基于核酸的疫苗和抗肿瘤分子（例如小干扰RNA），用于治疗和预防人类癌症疾病。我们最近表明，减毒沙门氏菌有效地将抗 HCMV RNase P 核酶序列传递至人类细胞，导致大量核酶表达并有效抑制病毒感染。此外，我们还生成了新型减毒沙门氏菌菌株，可有效递送 RNase P 核酶序列以在细胞中表达。然而，目前人们对沙门氏菌如何实现有效基因转移以在人类细胞中递送基于核酸的试剂（例如 RNase P 核酶）的机制知之甚少。目前尚未报道沙门氏菌介导的 RNase P 核酶递送是否在动物模型中特异性且有效地抑制病毒感染和发病机制。为了解决这些问题，我们建议首先研究生成的沙门氏菌菌株如何实现有效的基因转移。然后，我们将研究生成的沙门氏菌靶向递送 RNase P 核酶是否会消除培养的人类细胞中的 HCMV 基因表达和生长。使用小鼠巨细胞病毒（MCMV）感染作为模型系统，我们将确定所产生的沙门氏菌菌株是否能够高效地在动物体内靶向递送RNase P核酶，以及沙门氏菌介导的RNase P核酶的靶向递送是否在动物体内高效地靶向递送。阻断体内 MCMV 感染和发病机制。还将研究与产生的沙门氏菌菌株相关的潜在免疫/炎症反应和细胞毒性。我们的研究将产生具有高效基因转移活性的新型沙门氏菌菌株，可用于基因靶向临床应用。此外，这些结果将深入了解沙门氏菌如何在哺乳动物细胞中实现有效基因递送的机制，并将确定沙门氏菌介导的 RNase P 核酶递送是否特异性且有效地关闭培养细胞和动物中的基因表达。这项研究将促进沙门氏菌介导的 RNase P 核酶递送的发展，作为治疗 HCMV 和其他艾滋病相关病毒感染的新治疗方法。公共健康相关性：拟议的研究旨在开发新的药物，用于抗病毒分子的靶向基因传递，以治疗艾滋病相关病毒引起的感染，包括人类巨细胞病毒（HCMV），它是艾滋病患者最常见的机会性感染之一。我们的研究将促进一种新方法的开发，该方法可用作研究和治疗与人类病毒（如 HCMV）相关的感染的研究工具和治疗方法。