尿微囊泡和外泌体的蛋白质组学分析在多囊肾病和常染色体显性多囊肾病中的作用。

Proteomic Analysis of Urinary Microvesicles and Exosomes in Medullary Sponge Kidney Disease and Autosomal Dominant Polycystic Kidney Disease.

机构信息

Division of Nephrology, Dialysis, and Transplantation, Laboratory of Molecular Nephrology.

Renal Unit, Department of Medicine, University Hospital of Verona, Verona, Italy; and.

出版信息

Clin J Am Soc Nephrol. 2019 Jun 7;14(6):834-843. doi: 10.2215/CJN.12191018. Epub 2019 Apr 24.

DOI:10.2215/CJN.12191018

PMID:31018934

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6556712/

Abstract

BACKGROUND AND OBJECTIVES

Microvesicles and exosomes are involved in the pathogenesis of autosomal dominant polycystic kidney disease. However, it is unclear whether they also contribute to medullary sponge kidney, a sporadic kidney malformation featuring cysts, nephrocalcinosis, and recurrent kidney stones. We addressed this knowledge gap by comparative proteomic analysis.

DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The protein content of microvesicles and exosomes isolated from the urine of 15 patients with medullary sponge kidney and 15 patients with autosomal dominant polycystic kidney disease was determined by mass spectrometry followed by weighted gene coexpression network analysis, support vector machine learning, and partial least squares discriminant analysis to compare the profiles and select the most discriminative proteins. The proteomic data were verified by ELISA.

RESULTS

A total of 2950 proteins were isolated from microvesicles and exosomes, including 1579 (54%) identified in all samples but only 178 (6%) and 88 (3%) specific for medullary sponge kidney microvesicles and exosomes, and 183 (6%) and 98 (3%) specific for autosomal dominant polycystic kidney disease microvesicles and exosomes, respectively. The weighted gene coexpression network analysis revealed ten modules comprising proteins with similar expression profiles. Support vector machine learning and partial least squares discriminant analysis identified 34 proteins that were highly discriminative between the diseases. Among these, CD133 was upregulated in exosomes from autosomal dominant polycystic kidney disease and validated by ELISA.

CONCLUSIONS

Our data indicate a different proteomic profile of urinary microvesicles and exosomes in patients with medullary sponge kidney compared with patients with autosomal dominant polycystic kidney disease. The urine proteomic profile of patients with autosomal dominant polycystic kidney disease was enriched of proteins involved in cell proliferation and matrix remodeling. Instead, proteins identified in patients with medullary sponge kidney were associated with parenchymal calcium deposition/nephrolithiasis and systemic metabolic derangements associated with stones formation and bone mineralization defects.

PODCAST

This article contains a podcast at https://www.asn-online.org/media/podcast/CJASN/2019_04_24_CJASNPodcast_19_06_.mp3.

摘要

背景与目的

微泡和外泌体参与常染色体显性多囊肾病的发病机制。然而，它们是否也导致了髓质海绵肾的发生尚不清楚，髓质海绵肾是一种散发性肾脏畸形，其特征为囊肿、肾钙质沉着症和复发性肾结石。我们通过比较蛋白质组学分析来解决这一知识空白。

设计、地点、参与者和测量方法：通过质谱法测定了 15 例髓质海绵肾患者和 15 例常染色体显性多囊肾病患者尿液中分离的微泡和外泌体的蛋白质含量，然后进行加权基因共表达网络分析、支持向量机学习和偏最小二乘判别分析，以比较图谱并选择最具鉴别力的蛋白质。蛋白质组学数据通过 ELISA 进行验证。

结果

共从微泡和外泌体中分离出 2950 种蛋白质，其中 1579 种（54%）在所有样本中均被鉴定出来，但只有 178 种（6%）和 88 种（3%）特异性存在于髓质海绵肾微泡和外泌体中，183 种（6%）和 98 种（3%）特异性存在于常染色体显性多囊肾病微泡和外泌体中。加权基因共表达网络分析显示，包含具有相似表达谱的蛋白质的十个模块。支持向量机学习和偏最小二乘判别分析确定了 34 种在两种疾病之间具有高度鉴别力的蛋白质。其中，CD133 在常染色体显性多囊肾病患者的外泌体中上调，并通过 ELISA 验证。

结论

我们的数据表明，与常染色体显性多囊肾病患者相比，髓质海绵肾患者的尿液微泡和外泌体具有不同的蛋白质组学特征。常染色体显性多囊肾病患者尿液的蛋白质组学特征富含参与细胞增殖和基质重塑的蛋白质。相反，在髓质海绵肾患者中鉴定出的蛋白质与实质钙沉积/肾结石以及与结石形成和骨矿物质化缺陷相关的系统性代谢紊乱有关。