Genetic and hormonal mechanisms mediating sex differences in TB and TB-HIV

介导结核病和结核病艾滋病毒性别差异的遗传和激素机制

• 批准号: 10557906
• 负责人: WILLIAM Ramses BISHAI
• 金额: $ 81.38万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10557906
• 关键词: Abbreviations; Adoptive Transfer; Adrenal Glands; Androgens; Animal Model; Animals; Antitubercular Agents; Basic Science; Biological; Biological Factors; Bone Marrow; Cells; Country; Disasters; Dose; Epidemic; Epigenetic Process; Estrogens; Evaluation; FOXP3 gene; Female; Four Core Genotypes; Genes; Genetic; Genotype; Gonadal Steroid Hormones; Gonadal structure; Growth; HIV; HIV Infections; HIV/TB; Health Services Accessibility; Hormonal; Hormones; Hour; Human; Immune; Immune response; Immunity; Immunologics; Infection; Interferon Type II; Interleukin-4; Knockout Mice; Leukocytes; Link; Macrophage; Macrophage Activation; Mediating; Methods; Modeling; Molecular; Molecular Analysis; Mus; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; Necrotic Lesion; Observational epidemiology; Outcome; Pathogenesis; Patients; Peripheral Blood Mononuclear Cell; Persons; Play; Ploidies; Process; Processed Genes; Proliferating; Research Personnel; Resistance; Risk; Role; Sampling; Sampling Studies; Selective Estrogen Receptor Modulators; Severities; Sex Bias; Sex Chromosomes; Sex Differences; Signal Transduction; T cell response; T-Lymphocyte; TLR7 gene; TLR8 gene; TNF gene; TNFSF5 gene; Testosterone; Tuberculosis; Whole Blood; X Chromosome; X Inactivation; cytokine; dehydroepiandrosterone; design; genetic analysis; human female; human male; immune function; immunoregulation; male; mouse model; multidisciplinary; neglect; novel; pharmacologic; response; sex; tool; transcriptomics; tuberculosis treatment

PROJECT SUMMARY Tuberculosis (TB) occurs more frequently in males (~70% of cases) than females. While reduced access to care for females and undercounting of female cases may contribute modestly to this longstanding epidemiologic observation in some countries, both human studies (the Lűbeck disaster and eunuch studies) as well as animal models underscore the fact that biological factors play a dominant role in female resistance to TB. Despite the importance of this sex-bias in TB, it has gone largely neglected by basic science researchers. The central scientific premise of this application is that defining the biological mechanisms of the male bias for TB will enlighten our mechanistic understanding of TB pathogenesis and protective human immune responses in TB. A multidisciplinary team with decades of expertise on sex differences (Klein), TB pathogenesis (Bishai), and sex differences in HIV (Scully) will conduct the project. This proposal will investigate the impact of genetics (X chromosome complement) and sex steroid hormones on TB pathogenesis using cellular tools, animal models, and human samples. A key tool used in Aim 1 will be the novel four core genotype (FCG) mouse model, in which animals with XX genotypes have male gonads/hormone levels and those with XY genotypes have female gonads/hormone levels. This model will enable us to differentiate the impact of genetics from those of the sex steroids on murine control of TB. The X chromosome encodes numerous genes of immunologic importance including the genes that encode for TLR7, TLR8, CYBB, NEMO, CD40L, and FOXP3. The process of X-chromosome inactivation (XCI)--in which one female X chromosome is epigenetically silenced--is designed to provide balanced gene dosing between females and males. Certain genes, however, can escape XCI, leading to a double gene dose in females. The process of gene escape from XCI has not been studied in the context of TB, and advanced molecular tools will be used in Aim 2 to investigate gene escape from XCI as a cause of the male bias in severity of TB. While it is widely known that sex steroid signaling modulates immune function, the impact of sex steroids as a basis for the male bias in TB has not been thoroughly investigated. Testosterone has an immunotolerizing effect, reducing levels of IFN-γ and elevating levels of IL-4. In contrast, estrogen promotes higher levels of macrophage activation and increases in TNF-α levels. In Aim 2, we will carefully dissect the role of sex steroids using adoptive transfer methods as well as gonadectomized mice with selective hormone replacement. Lastly, in Aim 3 we will extend these studies to assess the intersection of HIV infection, a critical risk for TB, and sex differences in immunity. We will assess immune responses in samples from people living with HIV (PLWH) on ART versus healthy controls (HCs), specifically balancing groups for sex. We will evaluate the ability of whole blood and hMDM from PLWH and HCs to contain Mtb growth both alone and in the presence of DHEA. Lastly, we will assess transcriptomic responses in PBMCs from male and female PLWH and HCs following Mtb infection.

项目概要 结核病 (TB) 在男性（约占病例的 70%）中比女性更常见。虽然获得护理的机会减少 对于女性来说，女性病例的低估可能对这种长期存在的流行病学造成一定程度的影响 一些国家的观察，包括人类研究（吕贝克灾难和太监研究）以及动物研究 模型强调了这样一个事实：生物因素在女性对结核病的抵抗力中发挥着主导作用。尽管 虽然这种性别偏见在结核病中的重要性，但它在很大程度上被基础科学研究人员忽视了。中央 该应用的科学前提是，定义男性结核病偏向的生物学机制将 启发我们对结核病发病机制和结核病中保护性人类免疫反应的机制理解。一个 多学科团队在性别差异 (Klein)、结核病发病机制 (Bishai) 和性别方面拥有数十年的专业知识 艾滋病毒差异（Scully）将主持该项目。 该提案将调查遗传学（X 染色体补体）和性类固醇激素的影响 使用细胞工具、动物模型和人类样本研究结核病发病机制。目标 1 中使用的一个关键工具是 新型四核心基因型 (FCG) 小鼠模型，其中具有 XX 基因型的动物具有雄性 性腺/激素水平和具有 XY 基因型的人具有女性性腺/激素水平。该模型将 使我们能够区分遗传学与性类固醇对小鼠结核病控制的影响。 X 染色体编码许多具有免疫学重要性的基因，包括编码 TLR7、TLR8、CYBB、NEMO、CD40L 和 FOXP3。 X染色体失活（XCI）的过程——其中 一条女性 X 染色体被表观遗传沉默——旨在提供平衡的基因剂量 女性和男性。然而，某些基因可以逃避 XCI，导致女性体内的基因剂量加倍。这 XCI 基因逃逸过程尚未在结核病背景下进行研究，先进的分子工具将 用于目标 2，调查 XCI 中的基因逃逸是导致 TB 严重程度男性偏倚的原因。 虽然众所周知，性类固醇信号调节免疫功能，但性类固醇作为一种药物的影响 结核病中男性偏向的基础尚未得到彻底调查。睾酮具有免疫耐受作用， 降低 IFN-γ 水平并升高 IL-4 水平。相反，雌激素促进巨噬细胞水平升高 激活并增加 TNF-α 水平。在目标 2 中，我们将使用过继性方法仔细剖析性类固醇的作用 移植方法以及选择性激素替代的去性腺切除小鼠。 最后，在目标 3 中，我们将扩展这些研究，以评估艾滋病毒感染（结核病的一个关键风险）和 免疫力的性别差异。我们将评估艾滋病毒感染者 (PLWH) 样本中的免疫反应 ART 与健康对照 (HC) 的比较，特别是平衡性别组。我们会评估整体的能力 来自 PLWH 和 HC 的血液和 hMDM 可以单独抑制 Mtb 生长，也可以在 DHEA 存在的情况下抑制 Mtb 生长。最后， 我们将评估 Mtb 感染后男性和女性 PLWH 和 HC 的 PBMC 的转录组反应。