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

• 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
• 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

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HCMV感染通常与胃肠炎，肺炎和威胁性视网膜炎有关。耐药性HCMV菌株的出现（例如Ganciclovir）提出了开发新药和新型策略来打击HCMV感染的新药物。这项研究的目的是开发沙门氏菌作为一种新型的基因转移载体，用于靶向抗HCMV RNase P核酶来阻断病毒感染。沙门氏菌的减毒菌株可以充当载体系统，用于递送真核表达载体，并已证明可以输送治疗剂，包括基于核酸的疫苗和抗肿瘤分子（例如，小型干扰RNA）以治疗人类疾病的治疗和预防。我们最近表明，衰减的沙门氏菌有效地将抗HCMV RNase P核酶序列传递给人类细胞，从而导致实质性的核酶表达和有效抑制病毒感染。此外，我们已经产生了新型的减毒沙门氏菌菌株，这些菌株有效地传递了RNase P核酶序列以表达细胞中的表达。然而，目前，关于沙门氏菌如何实现有效基因转移的机制知之甚少。尚未报道沙门氏菌介导的RNase P核酶的递送特异性且有效地抑制动物模型中的病毒感染和发病机理。为了解决这些问题，我们建议首先研究产生的沙门氏菌菌株如何实现有效的基因转移。然后，我们将研究是否通过产生的沙门氏菌靶向递送RNase P核酶，以废除培养的人类细胞中的HCMV基因表达和生长。使用小鼠的鼠巨细胞病毒（MCMV）感染作为模型系统，我们将确定生成的沙门氏菌菌株对于动物中RNase P核酶的靶向递送高度高，以及由Salmonella介导的RNase P核酶是否高效地阻断MCMV感染和病原体的基础。还将研究与生成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有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，这些结果将提供有关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核酶的递送是否在关闭培养细胞和动物中的基因表达方面是特异性且有效的。这项研究将有助于开发沙门氏菌介导的RNase P核酶作为一种新型治疗方法，用于治疗HCMV和其他与AIDS相关的病毒。 公共卫生相关性：拟议的研究是为了在治疗由艾滋病相关病毒引起的感染（包括人类巨细胞病毒（HCMV））引起的抗病毒分子的靶向基因递送的新型药物，这导致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，该方法可以用作研究工具，以及一种研究和治疗与HCMV等人类病毒相关的感染的治疗方法。