TARGETED siRNA DELIVERY AS AN ANTI-HIV MICROBICIDE

靶向 siRNA 递送作为抗 HIV 杀菌剂

基本信息

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
项目类别：
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

项目摘要

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.

作为抗 HIV 杀菌剂的靶向 siRNA 递送 PI：Derek M. Dykxhoorn 项目概要：人类免疫缺陷病毒（HIV）是一种高度致命的慢病毒，它会在较长的时间内破坏人体的免疫系统。宿主的适应性免疫系统使它们容易受到多种机会性感染。与大多数人不同病毒的基因组复制独立于宿主细胞的基因组，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病毒生产时，病毒基因组保持完整且有能力。因此，可以采取防止吸收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（例如阴道粘膜）传播的结果。交付 siRNA 特异性沉默 HIV 生命周期早期事件所需的宿主因子，并作用于淋巴细胞阴道粘膜可能被证明是保护个人免受艾滋病毒感染的有效手段，并可作为一种潜在的杀菌剂。 siRNA作为遗传基因的临床应用面临的主要挑战之一治疗是将 siRNA 递送至适当靶细胞类型的细胞质的能力。我们最近有开发了一种新型脂质纳米颗粒，该颗粒涂有识别整合素分子 LFA-1 的抗体广泛表达于淋巴细胞。这些免疫纳米颗粒将用于将 siRNA 递送至 HIV人源化小鼠模型阴道粘膜中存在淋巴细胞。鉴于高水平序列异质性、HIV-1 突变的倾向以及抗 HIV siRNA 无法靶向进入病毒 RNA 基因组并防止整合，需要替代治疗靶点来防止艾滋病毒的传播。 HIV生命周期早期事件所必需但可有可无的宿主因素细胞功能可能被证明是一种有效的治疗替代方案。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当沉默时可以抑制艾滋病毒整合。然而，这些因素需要进行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引入针对治疗相关宿主因子的 siRNA 作为抑制病毒的潜在手段的载体 HIV人源化小鼠模型中的感染。这些实验将为临床前基础工作提供基础对于开发有效的基于 RNAi 的抗 HIV 杀微生物剂来说是必要的。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Systemic Gene Silencing in Primary T Lymphocytes Using Targeted Lipid Nanoparticles.

使用靶向脂质纳米颗粒对原代 T 淋巴细胞进行系统性基因沉默。

DOI：
10.1021/acsnano.5b02796
发表时间：
2015-06-10
期刊：
ACS nano
影响因子：
17.1
作者：
S. Ramishetti;Ranit Kedmi;Meir Goldsmith;F. Leonard;A. Sprague;B. Godin;M. Gozin;P. Cullis;D. Dyk
通讯作者：
D. Dyk