Proteomic Analysis of Urinary Microvesicles and Exosomes in Medullary Sponge Kidney Disease and Autosomal Dominant Polycystic Kidney Disease
Proteomic Analysis of Urinary Microvesicles and Exosomes in Medullary Sponge Kidney Disease and Autosomal Dominant Polycystic Kidney Disease
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DOI：
10.2215/cjn.12191018
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发表时间：
2019-06-07
影响因子：
9.8
通讯作者：
Zaza, Gianluigi
中科院分区：
文献类型：
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作者：
Bruschi, Maurizio;Granata, Simona;Zaza, Gianluigi
背景与目的微囊和外切体参与了常染色体显性遗传性多囊肾病的发病过程。然而，目前尚不清楚它们是否也对延髓海绵肾有贡献，延髓海绵肾是一种以囊肿、肾钙质沉着和肾结石复发为特征的散发性肾畸形。我们通过比较蛋白质组分析来解决这一知识差距。设计、设置、参与者和测量从15名髓质海绵肾患者和15名常染色体显性多囊肾患者的尿液中分离出的微囊和外切体的蛋白质含量，通过加权的基因表达网络分析、支持向量机学习和偏最小二乘判别分析来确定BMASS光谱，以比较这些图谱并选择最具区分性的蛋白质。结果共分离到2950个蛋白质组分，其中1579个(54%)为肾海绵肾微泡和外小体所特有，其中183个(6%)和98个(3%)为常染色体显性遗传性多囊肾病微囊和外小体所特有。加权基因共表达网络分析揭示了由具有相似表达谱的蛋白质组成的10个模块。支持向量机学习和偏最小二乘判别分析确定了34个在疾病之间高度区分的蛋白质。其中，CD133在常染色体显性遗传性多囊肾病外显子中表达上调，并经ELISA验证。结论与常染色体显性遗传性多囊肾病患者相比，髓质海绵肾患者的尿微泡和外显子的蛋白质组学特征不同。常染色体显性遗传性多囊肾病患者的尿蛋白组学分析显示，与细胞增殖和基质重塑有关的蛋白质丰富。相反，在髓质海绵肾患者中发现的蛋白质与实质性钙沉积/肾结石以及与结石形成和骨矿化缺陷相关的全身代谢紊乱有关。
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 theurine of 15patientswithmedullarysponge kidney and15patientswithautosomaldominant polycystic kidney diseasewasdeterminedbymass spectrometryfollowedbyweightedgene coexpressionnetworkanalysis, 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.ResultsAtotal of 2950 proteinswere isolated frommicrovesicles and exosomes, including 1579 (54%) identified in all samples but only 178 (6%) and 88 (3%) specific formedullary sponge kidneymicrovesicles 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 34proteins thatwere highlydiscriminativebetweenthediseases. Amongthese, CD133wasupregulated 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 comparedwith patientswith 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.