Proteomics in Type 1 Diabetes and its Complications

描述(由申请人提供)： 1型糖尿病(T1 DM)及其并发症，特别是糖尿病肾病(DN)是严重的公共卫生问题，发病率和死亡率都很高。糖尿病肾病是导致肾功能衰竭的主要原因，在其自然病史的大部分时间里都是默默无闻的，其发病机制也不完全清楚。皮肤成纤维细胞(SF)的体外行为反映了糖尿病肾病的风险。我们的微阵列基因表达数据显示，在快速与缓慢发展的糖尿病肾病患者(PTS)中，线粒体(MT)氧化磷酸化(OXPHOS)途径的基因表达水平在统计学上高度显著增加，这与氧化应激增加是一致的。鉴于基因和蛋白质的表达水平可能不同，本提案中的目标1将使用同位素编码的亲和标签和二维柱层析方法以及对蛋白质组数据的探索性视觉分析，测试快速与缓慢肾病发展的TIDM患者SF中mt蛋白质组途径的方向差异。我们的初步芯片数据还表明，没有糖尿病肾病的TIDM PTS在统计学上高度显著地增加了OXPHOS、电子传递复合体II、III和IV以及三羧酸(TCA)循环途径中基因的mt途径表达水平。其他研究发现，TIDM患者的非糖尿病一级亲属有证据表明氧化应激增加。AIM 2与TIDM本身的发病机制有关，并将检测TIDM(慢型糖尿病肾病)患者与非糖尿病无关对照(UC)以及这些UC与TIDM患者的一级亲属的mt方向通路蛋白表达。这些研究将确定在不复杂的TIDM患者中，是否存在与氧化应激相关的途径中mt蛋白表达增加，以及这是否可能是遗传决定的。这些研究可以提供糖尿病肾病和TIDM风险的替代标记物，并为TIDM及其肾脏并发症的发病机制提供洞察力。

DESCRIPTION (provided by applicant): Type 1 diabetes mellitus (T1DM) and its complications, especially diabetic nephropathy (DN) represent large public health problems with substantial morbidity and mortality. DN, the leading cause of kidney failure is silent through most of its natural history and DN pathogenesis is incompletely understood. Skin fibroblasts (SF) in vitro behaviors reflect DN risk. Our microarray gene expression data showed highly statistically significantly increased gene expression levels in the mitochondrial (mt) oxidative phosphorolation (OXPHOS) pathway in patients (pts) with rapid vs slow DN development, consistent with increased oxidative stress. Given that gene and protein expression levels may differ, Aim 1 in this proposal will test for mt proteome pathway directional differences in SF of TIDM pts with rapid vs slow DN development using isotope-coded affinity tag and 2-dimensional column chromatography methods and exploratory visual analyses of proteomic data. Our preliminary microarray data also indicated that TIDM pts without DN had highly statistically significantly increased mt pathway expression levels for genes in the OXPHOS, electron transport complexes II, III and IV and tricarboxylic acid (TCA) cycle pathways. Other studies found that non-diabetic first-degree relatives of TIDM pts have evidence of increased oxidative stress. Aim 2 is relevant to the pathogenesis of TIDM per se and will examine mt directional pathway protein expression as outlined above in TIDM (slow DN) pts vs non-diabetic unrelated controls (UC) as well as these UC vs first-degree relatives of TIDM pts. These studies will determine if there is increased mt protein expression in pathways relevant to oxidative stress in uncomplicated TIDM pts and whether this may be genetically determined. These studies could provide surrogate markers of DN and TIDM risk and provide insights into the pathogenesis of TIDM and its renal complications.