TARGETED siRNA DELIVERY AS AN ANTI-HIV MICROBICIDE

靶向 siRNA 递送作为抗 HIV 杀菌剂

• 批准号: 8880097
• 负责人: Derek Michael Dykxhoorn
• 金额: $ 40.63万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-15 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8880097
• 关键词: Anti-Retroviral Agents; Antibodies; Biological Models; CD4 Positive T Lymphocytes; Cell Line; Cell physiology; Cells; Clinical; Complement; Cytoplasm; Dendritic Cells; Dependency; Development; Disease; Drug Targeting; Ensure; Event; Exploratory/Developmental Grants Phase II; Gene Silencing; Gene Silencing Pathway; Gene Targeting; Genes; Genetic Materials; Genome; HIV; HIV Long Terminal Repeat; HIV vaccine; HIV-1; Heterogeneity; Heterosexuals; Immune system; Immunity; Individual; Infection; Infection prevention; Integration Host Factors; Integrins; Left; Life Cycle Stages; Lipids; Liposomes; Lymphocyte; Mammalian Cell; Mediating; Methods; Mucous Membrane; Mutate; Opportunistic Infections; Phase; Prevention; Process; Production; Proteins; RNA Interference; Reagent; Research; Safety; Satellite Viruses; Sexual Transmission; Small Interfering RNA; Small RNA; Staging; Structure; Subfamily lentivirinae; Surface; System; T-Lymphocyte; Testing; Therapeutic; Topical application; Vaccines; Vagina; Viral; Viral Genome; Virion; Virus; Virus Diseases; Virus Integration; Virus Replication; base; cell type; clinical application; efficacy testing; gene function; gene therapy; in vitro testing; in vivo; interest; macrophage; microbicide; mouse model; nanoparticle; novel; novel therapeutics; pathogen; pre-clinical; prevent; research study; screening; targeted treatment; therapeutic gene; therapeutic target; tissue culture; tool; transmission process; uptake; viral RNA

Targeted siRNA delivery as an anti-HIV microbicide PI: Derek M. Dykxhoorn Project Summary: Human immunodeficiency virus (HIV) is a highly lethal lentivirus which over a protracted course destroys the host's adaptive immune system leaving them vulnerable to numerous opportunistic infections. Unlike most viruses whose genome replicates independently of the host cell's genome, the HIV-1 genome integrates into and is replicated with the host genetic material. Therefore, even if therapeutic approaches can inhibit new virus production, the viral genome remains intact and competent. Therefore, strategies that can prevent the uptake and integration of the virus would be of tremendous clinical value. The vast majority of HIV infections occur as a consequence of viral transmission through mucosal surfaces, such as the vaginal mucosa. The delivery of siRNAs that specifically silence host factors required for early events in the HIV life cycle to lymphocytes in the vaginal mucosa could prove to be an effective means of protecting individuals from HIV infection and serve as a potential microbicide. One of the main challenges facing the clinical application of siRNAs as a genetic therapy is the ability to delivery siRNAs to the cytoplasm of the appropriate target cell types. We have recently developed a novel lipid nanoparticle that is coated with an antibody recognizing the integrin molecule LFA-1 which is broadly expressed on lymphocytes. These immuno-nanoparticles will be used to deliver siRNAs to lymphocytes present in the vaginal mucosa of humanized mouse models of HIV. Given the high level of sequence heterogeneity, the propensity of HIV-1 to mutate and the inability of anti-HIV siRNAs to target the incoming viral RNA genome and prevent integration, alternative therapeutic targets are required to prevent the transmission of HIV. Host factors that are necessary for early events in the HIV lifecycle but are dispensable for cellular functioning could prove to be an effective therapeutic alternative. Using a high-throughput RNA interference-based screening platform, we have identified a large number of potential therapeutic targets that could serve to inhibit HIV integration when silenced. However, these factors require extensive analysis and characterization to ensure their safety and efficacy. We will be combining the LFA-1-mediated cell-type specific vehicle to introduce siRNAs targeting therapeutically relevant host factors as a potential means to inhibit viral infection in humanized mouse models of HIV. These experiments will provide the preclinical groundwork necessary for the development of an effective RNAi-based anti-HIV microbicide.

靶向siRNA递送作为抗HIV杀微生物剂PI：Derek M. Dykxhoorn 项目摘要： 人类免疫缺陷病毒（HIV）是一种高致死性慢病毒，在一个漫长的过程中， 宿主的适应性免疫系统使它们容易受到许多机会性感染。不像大多数 对于基因组独立于宿主细胞基因组复制的病毒，HIV-1基因组整合到 并与宿主遗传物质一起复制。因此，即使治疗方法可以抑制新的病毒， 在生产过程中，病毒基因组保持完整和有能力。因此，可以防止摄取的策略 病毒的整合将具有巨大的临床价值。绝大多数艾滋病毒感染发生在 病毒通过粘膜表面如阴道粘膜传播的结果。提供 特异性沉默HIV生命周期早期事件所需的宿主因子的siRNA， 阴道粘膜可以被证明是一种有效的手段，保护个人免受艾滋病毒感染，并作为 一种潜在的杀菌剂SiRNA作为遗传学药物临床应用面临的主要挑战之一 治疗的一个重要方面是将siRNA递送至适当靶细胞类型的细胞质的能力。我们最近 开发了一种新的脂质纳米颗粒，其包被有识别整合素分子LFA-1的抗体 其在淋巴细胞上广泛表达。这些免疫纳米颗粒将用于将siRNA递送至 HIV的人源化小鼠模型的阴道粘膜中存在的淋巴细胞。鉴于高水平的 序列异质性、HIV-1突变的倾向以及抗HIV siRNA不能靶向HIV-1， 进入病毒RNA基因组并阻止整合，需要替代治疗靶点来阻止病毒RNA基因组的整合。 艾滋病毒的传播。宿主因素是HIV生命周期中早期事件所必需的， 可以证明是一种有效的治疗方法。使用高通量RNA 基于干扰的筛选平台，我们已经确定了大量潜在的治疗靶点， 可以抑制艾滋病毒的整合。然而，这些因素需要广泛的分析， 以确保其安全性和有效性。我们将结合LFA-1介导的细胞类型特异性 载体引入靶向治疗相关宿主因子的siRNA作为抑制病毒感染的潜在手段 HIV的人源化小鼠模型中的感染。这些实验将提供临床前基础 这是开发有效的基于RNAi的抗HIV杀微生物剂所必需的。