Using miRNAs to elucidate the cellular sources of HIV-1

使用 miRNA 阐明 HIV-1 的细胞来源

• 批准号: 8789293
• 负责人: Wendy Jean Maury
• 金额: $ 25.31万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8789293
• 关键词: Acute; Address; Adverse effects; Antiviral Agents; Antiviral Therapy; Biology; CD4 Positive T Lymphocytes; Cells; Dengue Virus; Development; Engineering; Future; Genome; Goals; HIV; HIV Genome; HIV Infections; HIV-1; Hematopoietic; Human; Immune response; In Vitro; Individual; Infection; Knowledge; Lymphoid; Mediating; MicroRNAs; Modeling; Molecular Cloning; Monitor; Myelogenous; Pathogenesis; Peripheral Blood Lymphocyte; Phenotype; Population; Production; Recombinants; Regimen; Relative (related person); Research; Role; SCID-hu Mice; Sensitivity and Specificity; Site; Source; Specificity; Staging; T-Lymphocyte Subsets; Technology; Testing; Therapeutic; Tissues; Tonsil; Tropism; Viral; Viral Load result; Viral Pathogenesis; Viral load measurement; Viremia; Virus; Virus Diseases; Virus Latency; Virus Replication; cell type; cellular targeting; chemokine; clinically relevant; cytokine; defined contribution; expectation; in vivo; influenzavirus; innovative technologies; insight; loss of function; macrophage; monocyte; new technology; novel strategies; novel therapeutics; public health relevance; response; tissue culture; tool; treatment strategy; virus pathogenesis

DESCRIPTION (provided by applicant): The relative in vivo contribution of permissive cell populations to HIV-1 viremia and total virus load during acute and subsequent persistent infections is poorly understood. A better understanding of the contribution of tissue macrophages and CD4+ T cell subpopulations will allow a more complete picture of HIV pathogenesis and potentially stimulate the development of cell-specific antiviral approaches that are likely to decrease side effects that are associated with current antiviral therapies. To date, approaches to understand the impact of the different cellular compartments in vivo have been limited and laborious. The novel approach proposed to be developed here will allow the field to clearly define the contribution of these cells to virus load and cytokine/chemokine changes. In these tissue culture and explant studies, we propose to develop a new and innovative technology to control the tropism of HIV for the purposes of delineating the contribution of specific cell types for HIV infection and pathogenesis and provide proof-of-principal that when used in vivo can provide important insights into impact of the cellular reservoirs of HIV. We will engineer recombinant HIV strains that are restricted in either myeloid- or lymphoid-specific cultures by harnessing cell- specific expression of endogenous miRNAs. Through incorporation of perfect miRNA target sites into the HIV genome, expression of these HIVs are restricted in targeted cellular subtypes, thereby allowing us to ascertain the contribution of these cells by a loss of function phenotype. By restricting HIV replication in a variety of different hematopoietic cellular subsets, we will determine how this impacts both virus and host biology in the context of primary ex vivo infections.

描述（由申请方提供）：在急性和随后的持续感染期间，容许细胞群对HIV-1病毒血症和总病毒载量的相对体内贡献尚不清楚。更好地了解组织巨噬细胞和CD 4 + T细胞亚群的贡献将使我们能够更全面地了解HIV的发病机制，并可能刺激细胞特异性抗病毒方法的发展，这些方法可能会减少与当前抗病毒治疗相关的副作用。迄今为止，了解体内不同细胞区室的影响的方法有限且费力。本文提出的新方法将使该领域能够清楚地定义这些细胞对病毒载量和细胞因子/趋化因子变化的贡献。在这些组织培养和外植体研究中，我们建议开发一种新的创新技术来控制HIV的向性，以描述特定细胞类型对HIV感染和发病机制的贡献，并提供在体内使用时可以提供对HIV细胞库影响的重要见解的主要证据。我们将通过利用内源性miRNAs的细胞特异性表达来设计限制在骨髓或淋巴特异性培养物中的重组HIV毒株。通过将完美的miRNA靶位点整合到HIV基因组中，这些HIV的表达被限制在靶细胞亚型中，从而使我们能够确定这些细胞通过功能表型丧失的贡献。通过限制HIV在各种不同造血细胞亚群中的复制，我们将确定这在原发性离体感染的背景下如何影响病毒和宿主生物学。