Altered miRNA expression in mesangial cells in lupus mice

描述（由申请人提供）：摘要：系统性红斑狼疮（SLE）是一种自身免疫性疾病，其特征在于细胞核成分的自身抗体。尽管对SLE的发病机制的理解进行了数十年的广泛工作，但还没有显示出指示疾病发病机制的特异性生物标志物。此外，对遗传和环境病原体的广泛分析尚未揭示疾病发生的共同机制。microRNA（miRNAs）的发现引发了一系列关于miRNAs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细胞功能和病理。微阵列数据显示，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，并显示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NZB/W雌性小鼠的系膜细胞在肾小球肾炎的发生和进展中显示出改变的miRNA谱。为了验证我们的假设，我们提出了以下具体目标：具体目标1：确定随着年龄的增长，来自NZB/W雌性小鼠的系膜细胞中miRNA的表达谱。虽然很明显miRNA在细胞分化、造血和免疫系统调节中起关键作用，但改变的miRNA表达模式的相关性及其在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认识。我们的研究将：1。确定来自NZB/W小鼠的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的发展。 2.将miRNA表达的变化与肾脏的病理变化相关联，以详细说明miRNA表达如何与疾病活动性平行。 3.寻求建立一个作用，改变miRNA表达在狼疮，从而改变我们的理解模式，SLE疾病是如何启动。 4.确定miRNA表达谱是否可用作疾病活动的生物标志物。 5.为通过调控miRNA在细胞中的表达来开发靶向疾病的新方法提供依据。 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microRNA（miRNAs）是一种短的非编码RNA分子，通过与靶mRNA的30-非翻译区（30-UTR）的不完全碱基配对来抑制基因表达，这引发了一系列关于miRNA如何调控基因表达的研究。我们最近证明了miRNA在免疫细胞中的重要性，并显示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中的miRNA表达改变，其表达改变可以被测量以鉴定肾小球肾炎的起始和进展。为了验证我们的假设，我们建议定义的表达谱的miRNA在系膜细胞从NZB/W雌性小鼠，因为他们的年龄。 这些实验的完成将对系膜细胞的分子运作产生重要的认识。我们的研究将检测来自新鲜纯化的肾小球系膜细胞的miRNA表达，我们将详细描述miRNA随疾病进展的变化。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表达如何与疾病活动平行。如果我们能够确定miRNA表达改变在狼疮中的作用，这些研究将改变我们对SLE疾病如何启动的理解。

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.