Role of Estrogen Receptors in Lupus-Modulators of the Inflammatory Response

摘要 系统性红斑狼疮(SLE)是一种以自身抗体为特征的自身免疫性疾病 产生和免疫复合体介导的器官损伤。系统性红斑狼疮的一个更深刻的特征是 女性患病率为9：1，男性为9：1。系统性红斑狼疮性别差异的原因是 多因素，包括性激素本身及其受体。雌激素的作用主要通过其 受体，雌激素受体α和β(ER？/ER？)然而，雌激素也可以通过非受体发挥作用。 调节机制和实质上，内质网调节独立于雌激素的生理功能。在 在之前的资助期间，我们获得了ER和ER基因敲除的狼疮倾向MRL/LPR和NZM2410小鼠。在这两个地方 雌性ER？KOS出现蛋白尿和病理性肾脏疾病的情况显著减少，并具有 显著延长存活期，尽管血清自身抗体水平升高。这些发现引导我们 假设狼疮性肾炎中ER缺乏的主要影响是肾脏对 发炎。我们从B6小鼠体内分离出ER？KO和ER？KO系膜细胞，发现ER？KO 肾小球系膜细胞对TLR2、3和7配体的反应明显减弱。ER？的表达不影响 肾小球系膜细胞反应。基于这些发现，我们假设ER的缺失是 钝化系膜细胞对TLR3/7诱导的狼疮小鼠肾脏保护作用 发炎。我们认为这种ER？保护作用是雌激素不依赖的，并通过TLR3/7介导 诱导内质网磷酸化。为了验证这一假设，我们提出了以下具体目标： 1.用Sle1在体内检测ER？缺乏对狼疮发病机制的影响。 Sle3和Sle1/3同系小鼠及ER？KO和ER？WT小鼠的骨髓移植。 2.明确影响炎症反应的TLR/ER？相互作用的体外分子机制 评估内质网对TLR3/7激活通路和TLR3/7影响的系膜细胞反应 关于ER？的表达。 3.利用突变体ER？在体内和体外确定ER？影响TLR信号转导的机制 影响特定ER？功能的Knockin菌株允许在 免疫反应。 这些研究将为ER？缺乏影响狼疮疾病的机制提供新的见解 表达并进一步描述ER和TLR之间的相互作用可能部分地诱导炎症 女性在狼疮中占主导地位。

Abstract Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by autoantibody production and immune complex mediated organ damage. One of the more profound features of SLE is females having a 9:1 prevalence of disease over men. The cause of this gender difference in SLE is multifactorial, including the sex hormones themselves and their receptors. Estrogen acts primarily via its receptors, estrogen receptor alpha and beta (ER¿/ER¿). Estrogen can also act, however, through non-receptor mediated mechanisms and, in kind, the ERs mediate physiologic functions independent of estrogen. In the previous funding period, we derived ER¿ and ER¿ knockout lupus prone MRL/lpr and NZM2410 mice. In both strains, the female ER¿ KOs developed significantly less proteinuria and pathologic renal disease and had significantly prolonged survival, despite increased serum levels of autoantibodies. These findings led us to postulate that the primary impact of ER¿ deficiency in lupus nephritis was on the response of the kidney to inflammation. We derived ER¿ KO and ER¿ KO mesangial cells from B6 mice and found that ER¿ KO mesangial cells had a marked blunted response to TLR 2, 3 and 7 ligands. ER¿ expression had no effect on mesangial cell responses as tested. Based on these findings, we hypothesize that the lack of ER¿ is renal protective in female lupus mice by blunting the response of mesangial cells to TLR3/7 induced inflammation. We believe this ER¿ protective effect is estrogen independent and mediated via TLR3/7 induced phosphorylation of ER¿. To test this hypothesis, we propose the following Specific Aims: 1. Determine in vivo the effects of ER¿ deficiency on known mechanisms of lupus pathogenesis using sle1, sle3 and sle1/3 congenic mice and bone marrow transplantation of ER¿ KO and ER¿ WT mice. 2. Define the in vitro molecular mechanisms underlying TLR/ER¿ interactions that impact the inflammatory response in mesangial cells assessing the impact of ER¿ on the TLR3/7 activation pathways and TLR3/7 on ER¿ expression. 3. Define in vivo and in vitro the mechanisms by which ER¿ impacts TLR signaling utilizing mutant ER¿ knockin strains that affect specific ER¿ functions allowing delineation of specific ER¿ functions on the immune response. These studies will provide novel insight into the mechanisms by which ER¿ deficiency impacts lupus disease expression and further delineate the interaction between ER¿ and TLR induced inflammation that may partially underlie the female predominance in lupus.