Enhancement of HIV transmission by hormones and bacterial metabolites

激素和细菌代谢物增强艾滋病毒传播

• 批准号: 8588037
• 负责人: John Christian Tilton
• 金额: $ 48.76万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-13 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8588037
• 关键词: Address; Affect; Antibodies; Area; Bacteria; Bacterial Vaginosis; Bioinformatics; Biological; Biopsy; Blood capillaries; CD4 Positive T Lymphocytes; Cells; Clinical; Contraceptive Agents; Contraceptive methods; Depo Provera; Epidemic; Epigenetic Process; Epithelial; Estradiol; Estrogens; Female; Flow Cytometry; Genital system; Goals; HIV; HIV Infections; Heterosexuals; Histones; Hormonal; Hormones; Immunology; In Vitro; Individual; Infection; Irrigation; Label; Lactobacillus; Link; Liquid substance; Local Microbicides; Mass Fragmentography; Measurement; Measures; Mediating; Menstrual cycle; Metabolic; Microbe; Pathway interactions; Patients; Pattern; Phase; Predisposition; Progesterone; Progestins; Proteins; Proteomics; Reporter; Reproductive Health; Research; Research Project Grants; Resistance; Risk; Sampling; Specimen; Staging; Staining method; Stains; Synthetic Progestogens; System; T-Cell Activation; Technology; Testing; Therapeutic; Tissues; Vaccines; Vagina; Variant; Viral; Virus; Virus Replication; Volatile Fatty Acids; Woman; Work; base; capillary; cell type; cervicovaginal; cohort; garcinol; histone acetyltransferase; improved; in vivo; inhibitor/antagonist; metabolomics; microbial; multidisciplinary; new technology; next generation; next generation sequencing; novel; novel strategies; novel therapeutics; programs; protein expression; public health relevance; rRNA Genes; reproductive; transmission process; Address; Affect; Antibodies; Area; Bacteria; Bacterial Vaginosis; Bioinformatics; Biological; Biopsy; Blood capillaries; CD4 Positive T Lymphocytes; Cells; Clinical; Contraceptive Agents; Contraceptive methods; Depo Provera; Epidemic; Epigenetic Process; Epithelial; Estradiol; Estrogens; Female; Flow Cytometry; Genital system; Goals; HIV; HIV Infections; Heterosexuals; Histones; Hormonal; Hormones; Immunology; In Vitro; Individual; Infection; Irrigation; Label; Lactobacillus; Link; Liquid substance; Local Microbicides; Mass Fragmentography; Measurement; Measures; Mediating; Menstrual cycle; Metabolic; Microbe; Pathway interactions; Patients; Pattern; Phase; Predisposition; Progesterone; Progestins; Proteins; Proteomics; Reporter; Reproductive Health; Research; Research Project Grants; Resistance; Risk; Sampling; Specimen; Staging; Staining method; Stains; Synthetic Progestogens; System; T-Cell Activation; Technology; Testing; Therapeutic; Tissues; Vaccines; Vagina; Variant; Viral; Virus; Virus Replication; Volatile Fatty Acids; Woman; Work; base; capillary; cell type; cervicovaginal; cohort; garcinol; histone acetyltransferase; improved; in vivo; inhibitor/antagonist; metabolomics; microbial; multidisciplinary; new technology; next generation; next generation sequencing; novel; novel strategies; novel therapeutics; programs; protein expression; public health relevance; rRNA Genes; reproductive; transmission process

DESCRIPTION (provided by applicant): This proposal aims to improve our understanding of how variations in hormone levels and reproductive tract microbes contribute to HIV acquisition risk in women. These are significant areas of research because more than half of new HIV infections are acquired by women, predominantly through heterosexual transmission. The goal of the proposed research project is to define how hormonal variation during the normal menstrual cycle (endogenous hormones) and with DMPA contraceptives (synthetic progestin) and bacterial vaginosis affect protein expression in ectocervical CD4+ T lymphocytes and their susceptibility to fusion and productive infection with HIV. These studies are powered to allow determination of hormonal effects, microbiota effects and interaction effects between hormones and vaginal microbes. We have found that bacterial metabolites called short chain fatty acids (SFCAs) that are present in healthy cervicovaginal fluid and increase dramatically during BV, strongly heighten the susceptibility of CD4+ T cells to productive HIV infection through the inhibition of histone deacetylases (HDACs). Finally, we will investigate a novel strategy to reduce HIV transmission by making cells more resistant to HIV infection. The three overlapping areas to be investigated in this project are: 1. Determine the effects of hormone levels and BV on susceptibility and activation state of CD4+ T lymphocytes from ectocervical biopsies. Ectocervical CD4+ T cell susceptibility to fusion and productive infection by HIV will be directly measured using our combination reporter virus system by flow cytometry. Activation states of CD4+ T cells will be quantified by flow cytometry and label-free proteomics, an unbiased and powerful strategy for identifying specific activation pathways. Advanced bioinformatics will be used to identify specific activation pathways that contribute to HIV replication in ectocervical CD4+ T cells. 2. Quantify bacterial species and metabolites present in cervicovaginal lavage (CVL) samples and correlate metabolites with specific species or groups of bacteria. CVL samples will be analyzed for bacterial species and for bacterial metabolites-including short chain fatty acids (SCFAs) that are present in the healthy female reproductive tract and elevated in BV. We have recently found that SCFAs enhance HIV infection of CD4+ T cells via a mechanism involving inhibition of HDACs. Advanced bioinformatics will be used to identify correlations between bacteria and their products and ectocervical CD4+ T cell protein expression and susceptibility to HIV. 3. Determine the ability of garcinol to reduce the susceptibility of ectocervical CD4+ T cells to HIV ex vivo in women with different hormone patterns and cervicovaginal microbiota. Garcinol, a histone acetyltransferase inhibitor, reverses the effects of SCFA-mediated HDAC inhibition of HIV susceptibility in vitro, and actually reduces infection even in unstimulated cells. These findings suggest that garcinol, or similar compounds, may represent a novel strategy to reduce heterosexual transmission of HIV by inhibiting productive infection of ectocervical CD4+ T cells.

描述（由申请人提供）：该提案旨在提高我们对激素水平和生殖道微生物的变化如何促进女性艾滋病毒的风险。这些是重要的研究领域，因为超过一半的新艾滋病毒感染是由女性获得的，主要是通过异性传播。拟议的研究项目的目的是确定在正常月经周期（内源激素）以及DMPA避孕药（合成孕激素）和细菌性阴道病的荷尔蒙变异如何影响蛋白质的蛋白质表达，蛋白质表达在纤维化CD4+ T淋巴细胞中及其对HIV融合和生产性感染的敏感性。这些研究有助于确定激素和阴道微生物之间的激素作用，微生物群和相互作用的影响。我们发现，细菌代谢产物称为短链脂肪酸（SFCA），它们存在于健康的宫颈阴道液中，并在BV期间急剧增加，从而通过抑制组蛋白脱乙酰基酶（HDACS）来强烈提高CD4+ T细胞对生产性HIV感染的敏感性。最后，我们将通过使细胞对HIV感染具有更具耐药性来研究一种新的策略，以减少HIV传播。在该项目中要研究的三个重叠区域是：1。确定激素水平和BV对院子里活检中CD4+ T淋巴细胞的易感性和激活状态的影响。通过流式细胞仪，将直接使用我们的组合记者病毒系统直接测量艾滋病毒融合和生产性感染的室内CD4+ T细胞敏感性。 CD4+ T细胞的激活状态将通过流式细胞术和无标签蛋白质组学来量化，这是一种识别特定活化途径的无偏和强大的策略。晚期生物信息学将用于识别有助于邻域CD4+ T细胞中HIV复制的特定激活途径。 2。量化宫颈阴道灌洗液（CVL）样品中存在的细菌物种和代谢产物，并将代谢产物与特定物种或细菌组相关。将分析CVL样品的细菌种类和细菌代谢物，包括在健康的女性生殖道中存在的短链脂肪酸（SCFA），并在BV中升高。我们最近发现，SCFA通过涉及抑制HDAC的机制增强了CD4+ T细胞的HIV感染。晚期生物信息学将用于鉴定细菌及其产物之间的相关性以及外部CD4+ T细胞蛋白表达和对HIV的易感性。 3。确定藤黄酚在具有不同激素模式和子宫颈阴道菌群不同的女性中降低外宫CD4+ T细胞对HIV的易感性的能力。甲醇是一种组蛋白乙酰转移酶抑制剂，逆转了 SCFA介导的HDAC在体外抑制HIV敏感性，甚至在未刺激的细胞中也会降低感染。这些发现表明，藤黄酚或类似化合物可能代表了一种新的策略，以减少艾滋病毒的异性传播，通过抑制邻域CD4+ T细胞的生产性感染。