Molecular signatures of diabetic kidney disease

 描述(申请人提供)：糖尿病肾病(DKD)发生在30-40%的2型糖尿病患者中。糖尿病和估计肾小球滤过率降低(EGFR&lt；60ml/min/1.73m2)的人患终末期肾脏疾病(ESRD)以及心血管疾病和死亡的风险特别高。尽管加强血糖控制和肾素血管紧张素系统(RAS)抑制，进展为终末期肾病和死亡的比率仍然很高。针对DKD阶段的新疗法的开发需要更多地了解导致DKD进展的机制，特别是在疾病的这个阶段。这项建议的总体目标是确定与2型糖尿病患者从EGFR减少到ESRD进展相关的生物途径。我们建议利用最先进的、有针对性的定量蛋白质组学平台来量化来自12个具有生物学前景的候选途径的179个蛋白质的表达，然后从EGFR降低到ESRD或EGFR下降50%。这些途径之所以被选择，是因为它们得到了DKD动物模型的广泛支持，以及参与晚期人类DKD的证据，但以前没有在人类DKD的这个阶段进行评估。使用我们最近完成的人类选择反应监测图谱(一个能够对20,300个人类蛋白质组进行定量蛋白质组学的多肽库)的免疫分析和定量靶向分析，我们检测了上述12个候选途径中的4个途径的28个蛋白质的尿液浓度。我们的初步数据显示，其中25种蛋白质的浓度明显异常，这与他们在显性DKD时相应通路的预期失调是一致的。我们假设，在EGFR降低时，12个靶向通路中的一个子集处于失调状态，并与进展为终末期肾病有关。为了验证这一假设，我们提出了一项嵌套在慢性肾功能不全队列(CRIC)研究的2型糖尿病亚队列中的病例对照研究，随后在西雅图肾脏研究(SKS)中进行了验证。我们将比较CRIC参与者中来自12个关键DKD途径的179种蛋白的尿液浓度，这些患者患有糖尿病和EGFR降低，这些患者在随访期间发生或没有进展为ESRD或50%的EGFR丢失。我们将使用这些新的测量方法和相应的临床数据来验证以下假设：目的1.确定在糖尿病和EGFR减少的患者中与DKD进展相关的蛋白质和相应的通路。目的2.在第二个队列，西雅图肾脏研究(SKS)中，验证CRIC研究(AIM 1)中与DKD进展相关的蛋白质和通路。

 DESCRIPTION (provided by applicant): Diabetic kidney disease (DKD) occurs in 30-40% of people with type 2 diabetes. People with diabetes and reduced estimated glomerular filtration rate (eGFR <60 ml/min/1.73m2) are at particularly high risk of end-stage renal disease (ESRD) as well as cardiovascular disease and death. Despite intensive glycemic control and renin angiotensin system (RAS) inhibition, the rates of progression to ESRD and death remain high. Development of new therapies targeting this DKD stage requires greater understanding of the mechanisms causing DKD progression, particularly at this stage of disease. The overall goal of this proposal is to identify the biologic pathways that are associated with progression from reduced eGFR to ESRD in type 2 diabetes. We propose to utilize a state-of-the-art, targeted, quantitative proteomics platform to quantify expression of 179 proteins from 12 biologically promising candidate pathways prior to progression from reduced eGFR to incident ESRD or 50% drop in eGFR. These pathways are selected because they have extensive support from DKD animal models as well as evidence of involvement in advanced human DKD, but have not been previously evaluated at this stage of human DKD. Using immunoassays and quantitative targeted assays from our recently completed Human Selected Reaction Monitoring Atlas (a library of peptides enabling quantitative proteomics for 20,300 human proteins), we examined urine concentration of 28 proteins from four of the above 12 candidate pathways. Our preliminary data suggests that the concentrations of 25 of these proteins are markedly abnormal, consistent with expected dysregulation of their corresponding pathways at the time of overt DKD. We hypothesize that a subset of the 12 targeted pathways are dysregulated at the time of reduced eGFR and are associated with progression to ESRD. To test this hypothesis, we propose a case-control study nested within a type 2 diabetes sub cohort of the Chronic Renal Insufficiency Cohort (CRIC) Study, with subsequent validation in the Seattle Kidney Study (SKS). We will compare urine concentration of 179 proteins from 12 key DKD pathways in CRIC participants with diabetes and reduced eGFR who do or do not progress to incident ESRD or 50% loss of eGFR during follow-up. We will use these novel measurements and the corresponding clinical data to test the following hypotheses: Aim 1. To identify the proteins and corresponding pathways which are altered in people with diabetes and reduced eGFR and are associated with DKD progression. Aim 2. To validate the proteins and pathways associated with DKD progression in the CRIC study (Aim 1) in a second cohort, the Seattle Kidney Study (SKS).