A Pilot Trial Assessing Urinary Gene Expression Profiling with an mRNA Array for Diabetic Nephropathy

• Published in: PloS one Vol. 7; no. 5; p. e34824
• Main Authors: Zheng, Min, Lv, Lin-Li, Cao, Yu-Han, Liu, Hong, Ni, Jie, Dai, Hou-Yong, Liu, Dan, Lei, Xiang-Dong, Liu, Bi-Cheng
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 18.05.2012; Public Library of Science (PLoS)
• Subjects: Adult; Aged; Aged, 80 and over; Biology; Biomarkers; Biomarkers - urine; Clinical medicine; Connective tissue growth factor; Diabetes; Diabetes mellitus; Diabetic nephropathies; Diabetic Nephropathies - diagnosis; Diabetic Nephropathies - genetics; Diabetic Nephropathies - urine; Diabetic nephropathy; E-cadherin; Epidermal growth factor receptors; Female; Gene expression; Gene Expression Profiling; Genes; Genetic research; Growth factors; Hospitals; Humans; Kidney diseases; Kidney transplantation; Laboratories; Male; Mass Screening; Medical research; Medicine; Messenger RNA; Microarray Analysis; Middle Aged; Monocyte chemoattractant protein 1; Nephrology; Nephropathy; Patients; Pilot Projects; Polyamide-imides; Relative abundance; Reproducibility of Results; RNA, Messenger - urine; Rodents; Searching; Sediments; Sensitivity and Specificity; Studies; Tissue inhibitor of metalloproteinase 1; Type 2 diabetes; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0034824

The initiation and progression of diabetic nephropathy (DN) is complex. Quantification of mRNA expression in urinary sediment has emerged as a novel strategy for studying renal diseases. Considering the numerous molecules involved in DN development, a high-throughput platform with parallel detection of multiple mRNAs is needed. In this study, we constructed a self-assembling mRNA array to analyze urinary mRNAs in DN patients with aims to reveal its potential in searching novel biomarkers. mRNA array containing 88 genes were fabricated and its performance was evaluated. A pilot study with 9 subjects including 6 DN patients and 3 normal controls were studied with the array. DN patients were assigned into two groups according to their estimate glomerular rate (eGFR): DNI group (eGFR>60 ml/min/1.73 m(2), n = 3) and DNII group (eGFR<60 ml/min/1.73 m(2), n = 3). Urinary cell pellet was collected from each study participant. Relative abundance of these target mRNAs from urinary pellet was quantified with the array. The array we fabricated displayed high sensitivity and specificity. Moreover, the Cts of Positive PCR Controls in our experiments were 24±0.5 which indicated high repeatability of the array. A total of 29 mRNAs were significantly increased in DN patients compared with controls (p<0.05). Among these genes, α-actinin4, CDH2, ACE, FAT1, synaptopodin, COL4α, twist, NOTCH3 mRNA expression were 15-fold higher than those in normal controls. In contrast, urinary TIMP-1 mRNA was significantly decreased in DN patients (p<0.05). It was shown that CTGF, MCP-1, PAI-1, ACE, CDH1, CDH2 mRNA varied significantly among the 3 study groups, and their mRNA levels increased with DN progression (p<0.05). Our pilot study demonstrated that mRNA array might serve as a high-throughput and sensitive tool for detecting mRNA expression in urinary sediment. Thus, this primary study indicated that mRNA array probably could be a useful tool for searching new biomarkers for DN.