The Role of Estrogen Receptor Alpha Variant Size and Localization in Modulating TLR7-Induced Inflammation

雌激素受体 α 变体大小和定位在调节 TLR7 诱导的炎症中的作用

基本信息

批准号：
10571915
负责人：
Melissa A Cunningham
金额：
$ 39.66万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-14 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10571915
关键词：
AF2 Address Affect Age Anti-Inflammatory Agents Antigen-Antibody Complex Antiinflammatory Effect Autoantibodies Autoimmune Diseases Autoimmunity B-Lymphocytes Binding Biological Blood Vessels Bone Marrow Cell Line Cell membrane Cell physiology Cells Chromatin DNA Binding Data Dendritic Cells Deposition Development Disease Dose End stage renal failure Endosomes Environmental Exposure Epidemiology Estrogen Receptor alpha Estrogen Receptors Estrogens Event Female Genes Genetic Transcription Genomics Goals Gonadal Steroid Hormones Human Immune Immune Targeting Immune response In Vitro Incidence Inflammation Inflammatory Inflammatory Infiltrate Inflammatory Response Innate Immune Response Interferon Gamma Receptor Beta Chain Investigation Kidney Kidney Diseases Length Ligand Binding Domain Location Lupus Lupus Nephritis Macrophage Mediating Membrane Mus Mutant Strains Mice Nephritis Nuclear Pathway interactions Patients Peripheral Blood Mononuclear Cell Predisposition Property Protein Isoforms Proteins Proteinuria Puberty Race Reporting Research Risk Factors Role Sampling Selective Estrogen Receptor Modulators Serum Sex Bias Sex Chromosomes Signal Transduction Structure Systemic Lupus Erythematosus TLR7 gene Technology Testing Therapeutic Time Tissue-Specific Gene Expression Tissues Toll-like receptors Transactivation Variant Woman Work X Inactivation cell type differential expression digital experimental study genetic corepressor immunoregulation in vivo lupus prone mice male monocyte mutant new therapeutic target next generation non-genomic novel novel strategies novel therapeutics overexpression protective effect prototype receptor reproductive response sex sex disparity tool transcription factor

项目摘要

Systemic lupus erythematosus (SLE) is one of many autoimmune diseases that disproportionately affects females. Although many risk factors for lupus are identified: >170 genes including GPR174, myriad environmental exposures, and aberrant X chromosome inactivation, none of these sufficiently explain the steep rise in incidence of ADs at the time of puberty in a female-specific manner. Epidemiology suggests a major role for sex hormones and their receptors in autoimmune diseases. We previously showed that female lupus-prone mice, expressing only a short form of estrogen receptor alpha (ERα short), have significantly reduced renal disease and increased survival. Determining the mechanism of this protective effect, which is estrogen dependent, is the primary goal of this proposal. Of note, ERα-/- (null) lupus prone mice were not similarly protected. Combined, our data suggest that the presence of the short form of ERα confers protection, not the absence of full-length ERα. Others and we demonstrated a critical role for ERα in dendritic cell (DC) development and endosomal Toll-like receptor (TLR) responsiveness. Interestingly, the ERα expressed in ERα short mice is similar in structure to an endogenous ERα variant (ERα46) that lacks the same AF-1 domain, and differentially regulates gene transcription compared to full length ERα. Overexpressing ERα46 in vitro also modulates TLR- induced responses, relevant to this proposal. In the proposed study, we will further investigate the role of ERα short variants in modulating TLR7-induced immune responses, and determine whether genomic and/or non- genomic mechanisms of ERα short variant action are protective. Our overall hypothesis is that increasing expression of ERα short or ERα46 in immune cells will be anti-inflammatory, and that the ratio of ERα46 to ERα66 is decreased in lupus patients versus healthy controls. We also hypothesize that targeting immune cells with novel anti-inflammatory selective estrogen receptor modulators (SERMs) that alter ERα membrane signaling and/or ERα-induced transcription will uncouple estrogen-mediated anti-inflammatory responses from those impacting reproductive tissues. We will test our hypotheses by accomplishing these Specific Aims: 1) Overexpress ERα short or treat immune cells with novel SERMs (OBHS, PaPE) that select for anti-inflammatory properties of ERα and determine the effect on known TLR7-induced inflammatory endpoints, 2) Investigate effects of membrane-only ERα signaling vs. nuclear only ERα expression on TLR7-induced pathways in mice, and 3) Identify ERα variants in human monocyte-derived dendritic cells (mo-DCs) and B cells using droplet digital PCR and Iso-Seq technology, to determine whether ERα46 is differentially expressed in lupus patients vs. controls, potentially explaining a biologic difference in females predisposed to autoimmunity. These aims will allow us to determine if we can separate ERα's reproductive effects from its potentially modifiable immune effects as a therapeutic strategy, which if successful, will provide novel approaches to immune modulation in lupus and other immune mediated diseases, especially those with a significant sex bias.

系统性红斑狼疮 (SLE) 是多种自身免疫性疾病之一，对患者的影响尤为严重虽然狼疮的许多危险因素已被确定：> 170 个基因，包括 GPR174、无数。环境暴露和异常 X 染色体失活，这些都不足以解释陡峭的流行病学表明青春期 AD 发病率的上升具有重要作用。对于性激素及其受体在自身免疫性疾病中的作用，我们之前表明女性易患狼疮。仅表达短形式雌激素受体α（ERα短）的小鼠，肾功能显着降低确定这种保护作用的机制，即雌激素。值得注意的是，ERα-/-（无效）狼疮易感小鼠的情况并不相似。综合起来，我们的数据表明，ERα 的缩写形式的存在提供了保护，而不是 ERα。缺乏全长 ERα。其他人和我们证明了 ERα 在树突状细胞 (DC) 发育中的关键作用。和内体 Toll 样受体 (TLR) 反应性提示，ERα 短小鼠中表达的 ERα 是结构与缺乏相同 AF-1 结构域的内源 ERα 变体 (ERα46) 相似，并且不同与全长 ERα 相比，其可调节基因转录体外过表达 ERα46 还可调节 TLR-。与本提案相关的诱导反应在拟议的研究中，我们将进一步研究 ERα 的作用。调节 TLR7 诱导的免疫反应的短变异，并确定基因组和/或非 ERα 短变异作用的基因组机制是保护性的。免疫细胞中表达ERα短或ERα46就会有抗炎作用，而且ERα46的比例与健康对照组相比，狼疮患者的 ERα66 减少。具有改变 ERα 的新型抗炎选择性雌激素受体调节剂 (SERM) 的免疫细胞膜信号传导和/或 ERα 诱导的转录将解开雌激素介导的抗炎作用我们将通过完成这些来检验我们的假设。具体目标： 1) 过度表达 ERα 或用新型 SERM（OBHS、PaPE）治疗免疫细胞，这些 SERM 会选择 ERα 的抗炎特性并确定对已知 TLR7 诱导的炎症终点的影响， 2) 研究仅膜 ERα 信号传导与仅细胞核 ERα 表达对 TLR7 诱导的影响小鼠体内的通路，以及 3) 识别人单核细胞衍生的树突状细胞 (mo-DC) 和 B 细胞中的 ERα 变体利用液滴数字PCR和Iso-Seq技术，确定ERα46在狼疮患者与对照组的比较，可能解释了易患自身免疫的女性的生物学差异。这些目标将使我们能够确定是否可以将 ERα 的生殖影响与其潜在的可改变性区分开来。免疫效应作为一种治疗策略，如果成功，将为免疫提供新的方法狼疮和其他免疫介导疾病的调节，特别是那些具有显着性别偏见的疾病。