TARGETED siRNA DELIVERY AS AN ANTI-HIV MICROBICIDE

靶向 siRNA 递送作为抗 HIV 杀菌剂

• 批准号: 8071962
• 负责人: Derek Michael Dykxhoorn
• 金额: $ 17.63万
• 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 2012-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8071962
• 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 Delivery Systems; 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; Screening procedure; 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; therapeutic gene; therapeutic target; tissue culture; tool; transmission process; uptake; viral RNA

DESCRIPTION (provided by applicant): 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. Heterosexual transmission is the leading cause of new HIV infections in the world. A microbicide providing true intracellular immunity would make a significant contribution to controlling the spread of this deadly virus.

描述(申请人提供)：人类免疫缺陷病毒(HIV)是一种高度致命的慢病毒，它在漫长的过程中破坏宿主的适应性免疫系统，使他们容易受到许多机会性感染。与大多数病毒的基因组独立于宿主细胞基因组复制不同，HIV-1基因组整合到宿主遗传物质中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病毒的产生，病毒基因组仍然完好无损。因此，能够防止病毒的吸收和整合的策略将具有巨大的临床价值。绝大多数艾滋病毒感染是病毒通过粘膜表面传播的结果，如阴道粘膜。将特定沉默艾滋病毒生命周期早期事件所需宿主因子的siRNAs输送到阴道粘膜中的淋巴细胞，可能被证明是保护个人免受艾滋病毒感染的有效手段，并可能成为一种潜在的杀微生物剂。将siRNAs作为一种遗传疗法应用于临床所面临的主要挑战之一是如何将siRNAs运送到适当靶细胞类型的细胞质中。我们最近开发了一种新型的脂质纳米颗粒，它被一种识别整合素分子LFA-1的抗体包裹，该分子广泛表达于淋巴细胞上。这些免疫纳米颗粒将被用来将siRNA传递给存在于人源化HIV小鼠模型阴道黏膜中的淋巴细胞。鉴于高度的序列异质性、HIV-1变异的倾向以及抗HIV-1 siRNAs不能针对传入的病毒RNA基因组并阻止整合，需要替代的治疗靶点来防止艾滋病毒的传播。宿主因素对于艾滋病毒生命周期的早期事件是必要的，但对于细胞功能来说却是必不可少的，这可能被证明是一种有效的治疗替代方案。使用高通量基于RNA干扰的筛查平台，我们已经确定了大量潜在的治疗靶点，当沉默时，这些靶点可能会抑制HIV的整合。然而，这些因素需要广泛的分析和表征，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siRNAs，作为抑制人源化HIV小鼠模型中病毒感染的潜在手段。这些实验将为开发有效的基于RNAi的抗HIV微杀菌剂提供必要的临床前基础。异性传播是世界上新的艾滋病毒感染的主要原因。一种能够提供真正的细胞内免疫的杀微生物剂将对控制这种致命病毒的传播做出重大贡献。