Altered miRNA expression in mesangial cells in lupus mice

描述(申请人提供)：摘要：系统性红斑狼疮(SLE)是一种自身免疫性疾病，其特征是对细胞中的核成分产生自身抗体。尽管几十年来对SLE的发病机制进行了广泛的研究，但还没有一个特定的生物标志物表明疾病的发病机制。此外，对遗传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microRNAs(MiRNAs)引发了一系列关于miRNA如何调控基因表达的研究。大量证据表明，miRNA系统有助于调节多种正常的细胞功能和病理。基因芯片数据显示，与健康对照组相比，SLE患者外周血细胞中miRNA的表达发生了变化。我们最近已经证明了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。鉴于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将显示出改变的miRNA图谱。为了验证我们的假设，我们提出了以下特定目标：特定目标1：确定随年龄增长的NZB/W雌性小鼠系膜细胞中miRNA的表达谱。虽然miRNA在调节细胞分化、造血和免疫系统的调节中起着关键作用，但miRNA表达模式的改变及其在SLE中的作用尚不清楚。肾小球系膜细胞在狼疮性肾炎的发病机制中起重要作用。这些实验的完成将对系膜细胞的分子工作产生重要的洞察力。我们的研究将：1.确定新鲜纯化的NZB/W小鼠肾小球系膜细胞的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的表达如何与疾病活动平行。3.寻求确定在狼疮中改变miRNA表达的作用，从而改变我们对SLE疾病如何启动的理解。4.确定miRNA表达谱是否可用作疾病活动性的生物标志物。5.通过调控miRNA在细胞中的表达，为开发靶向疾病的新途径提供证据。 公共卫生相关性：对系统性红斑狼疮(SLE)的遗传学和环境病原体的广泛分析尚未揭示疾病发生的共同机制。最近发现的microRNAs(MiRNAs)是一种短的非编码RNA分子，通过与靶mRNAs的30个非翻译区(30-UTR)不完全碱基配对来抑制基因表达，从而引发了一系列关于miRNA如何调控基因表达的研究。我们最近已经证明了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。鉴于miRNA在炎症调控中的作用，我们假设NZB/W雌性小鼠肾小球系膜细胞中miRNA的表达发生了变化，这种变化可以用来识别肾小球肾炎的发生和发展。为了验证我们的假设，我们建议定义随年龄增长的NZB/W雌性小鼠系膜细胞中miRNA的表达谱。这些实验的完成将对系膜细胞的分子工作产生重要的洞察力。我们的研究将检测新鲜纯化的系膜细胞的miRNA表达，并详细说明miRNA随着疾病进展的变化。我们将把miRNA表达的变化与肾脏的病理变化联系起来，以详细说明miRNA的表达如何与疾病活动平行。如果我们能在狼疮中确定miRNA表达改变的作用，这些研究将改变我们对SLE疾病启动方式的理解。

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.