Altered miRNA expression in mesangial cells in lupus mice

摘要：系统性红斑狼疮（SLE）是一种自身免疫性疾病，其特征是细胞中针对核成分的自身抗体。尽管在了解SLE的发病机制方面进行了数十年的广泛工作，但还没有一种特定的生物标志物被证明可以指示疾病的发病机制。此外，遗传学和环境病原体的广泛分析尚未揭示疾病起始的共同机制。最近发现的microrna （miRNAs）引发了一系列关于miRNA如何调节基因表达的研究。大量证据表明，miRNA系统有助于调节各种正常细胞功能和病理。芯片数据显示，与健康对照相比，SLE患者外周血细胞中的miRNA表达发生了改变。我们最近证明了miRNA在免疫细胞中的重要性，并证明了雌激素对免疫细胞中miRNA表达的选择性调节。考虑到miRNA在炎症调节中的作用，我们假设来自NZB/W雌性小鼠的系膜细胞在肾小球肾炎的发生和发展中会显示出miRNA谱的改变。为了验证我们的假设，我们提出了以下特定目的：特定目的1：定义随着年龄增长，NZB/W雌性小鼠系膜细胞中miRNA的表达谱。虽然miRNA在细胞分化、造血和免疫系统的调节中起着至关重要的作用，但miRNA表达模式的改变及其在SLE中的作用的相关性尚不清楚。系膜细胞在狼疮性肾炎的发病机制中起关键作用。这些实验的完成将对系膜细胞的分子运作产生重要的见解。我们的研究将：1。确定来自NZB/W小鼠新鲜纯化的系膜细胞的miRNA表达是否与疾病进展相关或预测疾病进展。2. 将miRNA表达的变化与肾脏的病理变化联系起来，以详细说明miRNA表达如何与疾病活动平行。3. 寻求在狼疮中建立miRNA表达改变的作用，从而改变我们对SLE疾病如何启动的理解范式。4. 确定miRNA表达谱是否可作为疾病活动性的生物标志物。5. 通过操纵细胞中的miRNA表达，为开发靶向疾病的新方法提供证据。

DESCRIPTION (provided by applicant): Abstract: Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibodies to nuclear components in the cell. Despite decades of extensive work in the understanding of the etiopathogenesis of SLE, there has not been a specific biomarker shown to indicate disease pathogenesis. Additionally, extensive analysis of both genetics and environmental pathogens has yet to reveal a common mechanism for the initiation of disease. The recent discovery of microRNAs (miRNAs) has initiated a flurry of investigations into how miRNA may regulate gene expression. Substantial evidence exists that the miRNA system contributes to the regulation of a wide variety of normal cellular functions and pathologies. Microarray data has shown that miRNA expression in peripheral blood cells of SLE patients is altered compared to healthy controls. We have recently demonstrated the importance of miRNA in immune cells and have shown the selective regulation of miRNA expression in immune cells by estrogen. Given the role of miRNA in the regulation of inflammation, we hypothesize that mesangial cells from NZB/W female mice will show an altered miRNA profile in the initiation and progression of glomerulonephritis. To test our hypothesis we propose the following Specific Aim: Specific Aim 1: Define the expression profile of miRNA in mesangial cells from NZB/W female mice as they age. While it is clear that miRNA plays a critical role in the regulation of cellular differentiation, hematopoesis, and the regulation of the immune system, the relevance of altered miRNA expression patterns and its role in SLE is unclear. Mesangial cells play a critical role in the pathogenesis of lupus nephritis. The completion of these experiments will yield important insight of the molecular workings of mesangial cells. Our studies will: 1. Determine if miRNA expression from freshly purified mesangial cells from NZB/W mice correlates or predicts the development of disease progression. 2. Correlate the changes in miRNA expression to pathological changes in the kidney to detail how miRNA expression parallels disease activity. 3. Seek to establish a role for altered miRNA expression in lupus and thus shift the paradigm in our understanding of how SLE disease is initiated. 4. Determine if miRNA expression profiles can be used as a biomarker for disease activity. 5. Provide evidence to develop novel approaches for the targeting disease by manipulating miRNA expression in cells. PUBLIC HEALTH RELEVANCE: Extensive analysis of both genetics and environmental pathogens of systemic lupus erythematosus (SLE) has yet to reveal a common mechanism for the initiation of disease. The recent discovery of microRNAs (miRNAs) which are short noncoding RNA molecules that inhibit gene expression through incomplete base pairing with the 30- untranslated region (30-UTR) of target mRNAs have initiated a flurry of investigations into how miRNA may regulate gene expression. We have recently demonstrated the importance of miRNA in immune cells and have shown the selective regulation of miRNA expression in immune cells by estrogen. Given the role of miRNA in the regulation of inflammation, we hypothesize altered miRNA expression in mesangial cells from NZB/W female mice with undergo alterations of expression that can be measured to identify the initiation and progression of glomerulonephritis. To test our hypothesis, we propose to define the expression profile of miRNA in mesangial cells from NZB/W female mice as they age. The completion of these experiments will yield important insight of the molecular workings of mesangial cells. Our studies will examine the miRNA expression from freshly purified mesangial cells and we will detail alterations of miRNA with disease progression. We will correlate the changes in miRNA expression to pathological changes in the kidney to detail how miRNA expression parallels disease activity. If we can establish a role for altered miRNA expression in lupus, these studies will shift the paradigm in our understanding of how SLE disease is initiated.