Molecular mechanisms of estrogen receptor alpha modulating the inflammatory response in systematic lupus erythematosus

雌激素受体α调节系统性红斑狼疮炎症反应的分子机制

• 批准号: 9254202
• 负责人: Melissa A Cunningham
• 金额: $ 17.21万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-07 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9254202
• 关键词: AF2; Address; Affect; Age; Alpha Cell; American; Antigen-Antibody Complex; Applications Grants; Attention; Autoantibodies; Autoimmune Diseases; Autoimmunity; Backcrossings; Binding; Bioinformatics; Cells; Cellular biology; Co-Immunoprecipitations; DNA Binding Domain; Data; Dendritic Cells; Deposition; Development; Diagnosis; Disease; Environment; Estradiol; Estrogen Receptor alpha; Estrogens; Female; Gene Targeting; Genetic Transcription; Genomics; Goals; Grant; High Prevalence; Hormonal; Hormone Receptor; Human; Immune; Immunology; In Vitro; Incidence; Infertility; Inflammation; Inflammatory; Inflammatory Response; Innate Immune Response; Interferon Gamma Receptor Beta Chain; Interferon-alpha; Interleukin-1 beta; Interleukin-6; Kidney Diseases; Knock-out; Knockout Mice; Knowledge; Laboratories; Length; Ligands; Link; Lupus; Lupus Erythematosus; Mediating; Mentors; Mentorship; Minority; Molecular; Mouse Strains; Mus; N-terminal; Nuclear Hormone Receptors; Pathogenesis; Pathway interactions; Patients; Phenotype; Physiological; Production; Protein Isoforms; Public Health; RNA Interference; RNA Splicing; Regulation; Regulator Genes; Research; Research Personnel; Risk; Risk Factors; Role; Sampling; Serum; Sex Bias; Signal Pathway; Systemic Lupus Erythematosus; TLR7 gene; Testing; Therapeutic Agents; Toll-like receptors; Training; Variant; Woman; Work; career; cell type; experimental study; health disparity; improved; in vivo; inhibitor/antagonist; interleukin-23; knock-down; knockout animal; lupus-like; monocyte; mouse model; mutant; overexpression; peripheral blood; promoter; protective effect; public health relevance; response; sex; therapy development; translational scientist

 DESCRIPTION (provided by applicant): Ninety percent of those diagnosed with systemic lupus erythematosus (SLE) are women, with peak incidence between the ages of 15 and 45, when women are most hormonally active. Despite significant research effort, the mechanisms underlying this sex bias remain unclear. Our laboratory previously backcrossed estrogen receptor alpha knockout (ERαKO) mice onto the NZM2410 lupus prone background. We demonstrated that female NZM/ERαKO mice had significantly less renal disease and significantly prolonged survival compared to WT littermates despite similar serum autoantibodies and glomerular immune complex deposition. ERαKO mice are not ERα null, but rather express an N-terminally truncated ERα. They have physiologic deficiencies including infertility due to disruption of a critical activation domain (AF-1). We showed that dendritic cell (DCs) from NZM/ERαKO mice have a blunted inflammatory response to Toll-like receptor (TLR) ligands. When these mice were ovariectomized, the protective phenotype was lost. Upon estradiol-repletion, protection was restored. True ERα null mice are not protected, suggesting that estrogen in the presence of the AF-1 mutant confers protection, rather than the absence of the full-length ERα66. Interestingly, the truncated ERα expressed in the ERαKO animal is structurally similar to ERα46, an endogenous ERα splice variant that lacks the AF-1 domain, and is a negative regulator of gene transcription. ERα46 has an identical DNA binding domain to ERα66 and is a powerful inhibitor of ERα66. We hypothesize that ERα46 expression has a protective effect in lupus. The goal of this project is to determine the role of ERα46 in SLE and TLR-induced inflammation and to improve our understanding of ERα-mediated transcription in the setting of inflammation. In order to accomplish this, Dr. Cunningham will (1) Define the in vitro molecular mechanisms underlying ERα46- and ERα66-regulated transcriptional activity impacting the innate inflammatory response of murine DCs, (2) Determine ex vivo and in vivo the role of ERα46 in TLR-induced lupus utilizing a murine strain expressing an A/B truncation mutant of ERα (ERαAF10), and (3) Use human peripheral blood monocyte (PBMC)-derived DCs to define the role of ERα46 and ERα66 in regulation of the innate immune response in DCs from controls vs. lupus patients. Dr. Cunningham is a clinician-investigator with a long-term career goal of becoming an independent basic and translational researcher in the field of immunology with a focus on SLE. To facilitate her transition to independence, she seeks to further her training in dendritic cell biology, ChIPseq and bioinformatics, and the use of human cells from patient samples. Dr. Cunningham has a mentorship team with an outstanding mentoring track record and wide expertise to support her project. The studies proposed in this K08 mentored grant application will significantly advance the field by providing understanding of ERα-mediated transcription in the setting of TLR-induced inflammation, including ERα binding partners, gene targets, and modulation by ERα46, which represents a potential therapeutic agent.

 描述（由申请人提供）：90%的系统性红斑狼疮（SLE）患者为女性，发病高峰年龄为15至45岁，此时女性生殖活动最活跃。尽管有大量的研究工作，这种性别偏见的机制仍然不清楚。我们的实验室先前将雌激素受体α敲除（ERαKO）小鼠与NZM 2410狼疮易感背景小鼠进行回交。我们证明，尽管血清自身抗体和肾小球免疫复合物沉积相似，但与WT同窝小鼠相比，雌性NZM/ERαKO小鼠的肾脏疾病显著较少，生存期显著延长。ERαKO小鼠不是ERα null，而是表达N末端截短的ERα。他们有生理缺陷，包括由于关键激活结构域（AF-1）的破坏而导致的不育。我们发现NZM/ERαKO小鼠的树突状细胞（DCs）对Toll样受体（TLR）配体的炎症反应减弱。当这些小鼠卵巢切除后，保护性表型丧失。雌二醇补充后，保护作用恢复。真正的ERα缺失小鼠没有受到保护，这表明雌激素在AF-1突变体的存在下提供保护，而不是在全长ERα66缺失的情况下。有趣的是，ERαKO动物中表达的截短ERα在结构上与ERα46相似，ER α 46是一种缺乏AF-1结构域的内源性ERα剪接变体，是基因转录的负调节因子。ERα46具有与ERα66相同的DNA结合结构域，是ERα66的强效抑制剂。我们推测ERα46表达在狼疮中具有保护作用。本项目的目的是确定ERα46在SLE和TLR诱导的炎症中的作用，并提高我们对ERα介导的炎症转录的理解。为了实现这一目标，Cunningham博士将（1）确定ERα46和ERα66调节的转录活性影响小鼠DC先天性炎症反应的体外分子机制，（2）利用表达ERα A/B截短突变体（ERα AF 10）的小鼠品系，（3）利用人外周血单核细胞（PBMC）来源的DC来确定ERα46和ERα66在调节来自对照和狼疮患者的DC的天然免疫应答中的作用。Cunningham博士是一名临床研究员，其长期职业目标是成为免疫学领域的独立基础和转化研究人员，重点是SLE。为了促进她向独立的过渡，她寻求进一步她在树突状细胞生物学，ChIPseq和生物信息学，以及从患者样本中使用人类细胞的培训。坎宁安博士拥有一支拥有出色指导记录和广泛专业知识的指导团队来支持她的项目。在K 08指导的资助申请中提出的研究将通过提供对TLR诱导的炎症背景下ERα介导的转录的理解来显著推进该领域，包括ERα结合伴侣，基因靶点和ERα46的调节，ERα46代表了一种潜在的治疗剂。