Proteomics in Type 1 Diabetes and its Complications

描述（由申请人提供）： 1型糖尿病（T1DM）及其并发症，特别是糖尿病肾病（DN）是一个严重的公共卫生问题，具有较高的发病率和死亡率。DN是肾衰竭的主要原因，在其大部分自然史中是沉默的，并且DN发病机制不完全清楚。皮肤成纤维细胞（SF）的体外行为反映了DN的风险。我们的微阵列基因表达数据显示，在快速与缓慢DN发展的患者（pts）中，线粒体（mt）氧化磷酸化（OXPHOS）途径中的基因表达水平在统计学上显著增加，与氧化应激增加一致。鉴于基因和蛋白质表达水平可能不同，本提案的目标1将使用同位素编码的亲和标签和二维柱色谱法以及蛋白质组数据的探索性可视化分析，检测快速与缓慢DN发展的TIDM患者SF中mt蛋白质组通路方向差异。我们的初步微阵列数据还表明，TIDM患者没有DN有高度统计学显着增加MT途径的表达水平的基因在OXPHOS，电子传递复合物II，III和IV和三羧酸（TCA）循环途径。其他研究发现，TIDM患者的非糖尿病一级亲属有氧化应激增加的证据。目的2与T1DM本身的发病机制相关，并将在T1DM（缓慢DN）患者与非糖尿病无关对照（UC）以及这些UC与T1DM患者的一级亲属中检查如上所述的mt定向途径蛋白表达。这些研究将确定在无并发症的TIDM患者中，与氧化应激相关的通路中是否存在增加的mt蛋白表达，以及这是否可能由遗传决定。这些研究可以提供DN和TIDM风险的替代标志物，并为TIDM及其肾脏并发症的发病机制提供见解。

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.