Molecular mechanisms contributing to gender differences in regulatory T cell function in Crohn's disease and experimental IBD

克罗恩病和实验性 IBD 调节性 T 细胞功能性别差异的分子机制

• 批准号: 9146962
• 负责人: Wendy Ann Goodman
• 金额: $ 11.3万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-21 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9146962
• 关键词: Adoptive Transfer; Advisory Committees; Agonist; Antigens; Area; Autoimmune Diseases; Autoimmunity; Backcrossings; Binding; Biological Assay; Caring; Cell physiology; Cells; Chronic; Clinical; Complex; Coupled; Crohn' s disease; Data; Defect; Development; Development Plans; Diet; Environment; Environmental Risk Factor; Estradiol; Estrogen Receptor alpha; Estrogen Receptor beta; Estrogen Receptors; Estrogens; Estrous Cycle; Exhibits; Exogenous Factors; Faculty; Female; Flare; Frequencies; Functional disorder; Funding; Future; Gene Expression; Gene Targeting; Genes; Genetic; Goals; Grant; Health; Hormonal; Human; IL2RA gene; Ileitis; Immune; Immunoprecipitation; In Vitro; Incidence; Inflammation; Inflammatory Bowel Diseases; Intestinal Mucosa; Intestines; Intrinsic factor; Knockout Mice; Knowledge; Lymphocyte Function; Mediating; Mentorship; Messenger RNA; Modeling; Molecular; Mucous Membrane; Mus; Oral Contraceptives; Outcome; Outcome Measure; Ovarian hormone; Paper; Pathogenesis; Pathology; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Positioning Attribute; Protein Isoforms; Proteins; Puberty; Public Health; Regulatory T-Lymphocyte; Reporter; Research; Research Personnel; Resistance; Resources; Risk Factors; Role; Series; Serum; Sex Bias; Signal Transduction; Symptoms; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Time; Tissues; Training; Universities; Up-Regulation; Wild Type Mouse; base; career; career development; chromatin immunoprecipitation; design; experience; faculty research; gender difference; health related quality of life; improved; in vivo; instructor; male; meetings; microbial; mouse model; next generation; novel; novel therapeutics; personalized care; programs; research study; response; responsible research conduct; restoration; sex; steroid hormone; therapy design; transcription factor; transcriptome sequencing; treatment strategy

 DESCRIPTION (provided by applicant): Crohn's disease (CD) represents a significant public health challenge. Pathogenesis is promoted by chronic inflammation of the intestinal mucosa, resulting in loss of tolerance to exogenous factors such as microbial and dietary antigens. There is an increased incidence of CD among female patients, who also experience worse clinical symptoms compared to males. Genetic and environmental factors such as the ovarian hormone 17-β-estradiol (estrogen, E2) have been identified as risk factors for CD, given the frequent onset of females' CD during puberty, as well as females' worsening symptoms during times of elevated E2 levels. Recent exciting studies from our group demonstrated that E2-mediated signaling is impaired in T cells isolated from female ileitis-prone mice. This is in contrast to th effects of E2 signaling under non-pathological conditions, in which E2 induces conversion of conventional T cells (Tconv) to regulatory T cells Treg, thus enhancing immune protection. Thus, there is a need for mechanistic studies aimed at delineating the molecular mechanisms by which E2 may result in exacerbated CD. Studies proposed herein take advantage of a unique resource, a spontaneous murine model of Crohn's-like ileitis (SAMP1/YitFc (SAMP)), to clarify the mechanistic role of E2 signaling in experimental ileitis. Our group has recently shown that SAMP mice recapitulate several key aspects of human CD, including Treg dysfunction (Ishikawa et al, Mucosal Immunol 2013) and female sex bias (Goodman et al, Mucosal Immunol 2014). Specifically, we recently showed that Tregs isolated from female SAMP (SAMP-F) exhibit impaired suppressive function in vivo and reduced expression of key Treg-associated genes such as foxp3 and helios. Following treatment with exogenous E2, SAMP-M expand their mucosal Treg pool and demonstrate improvement in ileitis, whereas SAMP-F do not expand their Tregs and do not improve clinically. In exciting new data generated since the first submission, we have shown that gene expression of estrogen receptor beta (ERβ) is markedly upregulated in SAMP-F T cells, and that deletion of ERβ results in clinical improvement and a restoration of Treg function in SAMP-F mice. These novel observations serve as the basis for the central hypothesis that dysfunctional E2/ERβ signaling predicates female sex bias in SAMP, despite females' higher circulating levels of E2. The goals of this proposal are: (1) To demonstrate that dysregulated ERβ signaling contributes to worsened SAMP-F ileitis; and (2) to identify the molecular mechanism(s) responsible for dysfunctional Treg responses in SAMP-F. The successful completion of these aims will provide key knowledge about E2-dependent mechanism(s) for female sex bias in IBD. Given the immunoprotective role of E2 under non-pathological conditions, understanding the mechanisms by which E2 signaling may be subverted in female CD patients is essential for development of novel therapies designed to function effectively in different hormonal environments and for future efforts towards individualized care. The candidate's long-term career goals include the development of an independent research program in the scientific area of molecular control of lymphocyte function in IBD. She has extensive training in the pathogenesis of autoimmune disease, having authored three first-author research papers in this area and several reviews (see attached Candidate Biosketch). A rigorous Career Development Plan has been developed (see attached documents) which includes didactic coursework, participation in numerous research seminar series, training in the responsible conduct of research, and regular meetings with her Scientific Advisory Committee. The sponsor, Dr. Theresa Pizarro, is a well- established investigator in the field of IBD with a strong record of funding and mentorship. Dr. Pizarro, along with the Department of Pathology at Case Western Reserve University, have committed fully to supporting the candidate as she transitions to a fully independent research faculty position. Dr. Goodman will be promoted to the Faculty rank of Instructor within the 2014-2015 academic year, allowing her to compete for R-series grants that will derive from data and hypotheses generated in the current K01 proposal.

 描述（由申请人提供）：克罗恩病（CD）代表着重大的公共卫生挑战。肠粘膜的慢性炎症促进了发病机制，导致对微生物和饮食抗原等外源因素的耐受性丧失。女性患者的克罗恩病发病率有所增加，与男性患者相比，她们的临床症状也更严重。鉴于女性 CD 在青春期频繁发作，以及女性在 E2 水平升高期间症状恶化，遗传和环境因素（例如卵巢激素 17-β-雌二醇（雌激素，E2））已被确定为 CD 的危险因素。我们小组最近令人兴奋的研究表明，从易患回肠炎的雌性小鼠中分离出的 T 细胞中，E2 介导的信号传导受损。这与非病理条件下 E2 信号传导的作用相反，在非病理条件下，E2 诱导常规 T 细胞 (Tconv) 转化为调节性 T 细胞 Treg，从而增强免疫保护。因此，需要进行机制研究，旨在阐明 E2 可能导致 CD 恶化的分子机制。 本文提出的研究利用独特的资源，即克罗恩病样回肠炎的自发小鼠模型 (SAMP1/YitFc (SAMP))，来阐明 E2 信号传导在实验性回肠炎中的机制作用。我们的研究小组最近表明，SAMP 小鼠重现了人类 CD 的几个关键方面，包括 Treg 功能障碍（Ishikawa 等人，Mucosal Immunol 2013）和女性性别偏见（Goodman 等人，Mucosal Immunol 2014）。具体来说，我们最近表明，从雌性 SAMP (SAMP-F) 中分离出的 Tregs 表现出体内抑制功能受损，并且关键 Treg 相关基因（如 Foxp3 和 helios）的表达减少。用外源性 E2 治疗后，SAMP-M 扩大了其粘膜 Treg 库并显示出回肠炎的改善，而 SAMP-F 不会扩大其 Tregs 且临床上也没有改善。自首次提交以来产生的令人兴奋的新数据表明，SAMP-F T 细胞中雌激素受体 β (ERβ) 的基因表达显着上调，并且 ERβ 的缺失可导致 SAMP-F 小鼠的临床改善和 Treg 功能的恢复。这些新颖的观察结果为以下中心假设奠定了基础：尽管女性的 E2 循环水平较高，但 E2/ERβ 信号传导功能失调预示着 SAMP 中的女性性别偏见。该提案的目标是： (1) 证明 ERβ 信号传导失调会导致 SAMP-F 回肠炎恶化； (2) 确定导致 SAMP-F 中 Treg 反应功能失调的分子机制。这些目标的成功完成将为 IBD 女性性别偏见的 E2 依赖性机制提供关键知识。鉴于 E2 在非病理条件下的免疫保护作用，了解 E2 信号在女性 CD 患者中可能被破坏的机制对于开发旨在在不同激素环境中有效发挥作用的新疗法以及未来努力实现个体化护理至关重要。 该候选人的长期职业目标包括在 IBD 淋巴细胞功能分子控制科学领域开发独立研究项目。她在自身免疫性疾病发病机制方面接受过广泛的培训，在该领域撰写了三篇第一作者研究论文和几篇评论（参见随附的候选生物草图）。已经制定了严格的职业发展计划（见附件），其中包括教学课程、参与众多研究研讨会系列、负责任的研究行为培训以及与科学咨询委员会的定期会议。资助者 Theresa Pizarro 博士是 IBD 领域的知名研究者，在资助和指导方面拥有良好的记录。皮萨罗博士和凯斯西储大学病理学系已全力支持该候选人过渡到完全独立的研究教职职位。 Goodman 博士将在 2014-2015 学年晋升为讲师，从而使她能够竞争 R 系列补助金，这些补助金将来自当前 K01 提案中生成的数据和假设。