Urinary mRNA Expression of Glomerular Podocyte Markers in Glomerular Disease and Renal Transplant.

Silvia Armelloni,Roberta Cerutti,Gabrilella Moroni,Mirco Belingheri,Deborah Mattinzoli,Piergiorgio Messa,Carlo Alfieri,Patrizia Passerini,Donata Cresseri,Masami Ikehata

doi:10.3390/diagnostics11081499

Silvia Armelloni, Roberta Cerutti + Show 8 more

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https://doi.org/10.3390/diagnostics11081499

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Abstract

The research of novel markers in urinary samples, for the description of renal damage, is of high interest, and several works demonstrated the value of urinary mRNA quantification for the search of events related to renal disease or affecting the outcome of transplant kidneys. In the present pilot study, a comparison of the urine mRNA expression of specific podocyte markers among patients who had undergone clinical indication to renal transplanted (RTx, n = 20) and native (N, n = 18) renal biopsy was performed. The aim of this work was to identify genes involved in podocytes signaling and cytoskeletal regulation (NPHS1, NPHS2, SYNPO, WT1, TRPC6, GRM1, and NEUROD) in respect to glomerular pathology. We considered some genes relevant for podocytes signaling and for the function of the glomerular filter applying an alternative normalization approach. Our results demonstrate the WT1 urinary mRNA increases in both groups and it is helpful for podocyte normalization. Furthermore, an increase in the expression of TRPC6 after all kinds of normalizations was observed. According to our data, WT1 normalization might be considered an alternative approach to correct the expression of urinary mRNA. In addition, our study underlines the importance of slit diaphragm proteins involved in calcium disequilibrium, such as TRPC6.

Highlights

The need to obtain valuable, sensitive, and specific information to recognize the signs of renal dysfunction and how it contributes to identifying glomerular disease and renal transplant (RTx) decline has recently led to new investigations
With the intent to dwell on functional aspects of podocyte biology leading to injury, we considered some of the crucial genes relevant for podocyte signaling pathways and the function of the glomerular filter applying an alternative normalization approach
Among markers contained in the primer mix, we considered and measured in this paper the following genes (Supplementary Table S1: primer sequences): NPHS1, NPHS2, Wilms’ tumor suppressor 1 (WT1), SYNPO, TRPC6, GRM1, neurogenic differentiation factor one (NEUROD)

Summary

Introduction

The need to obtain valuable, sensitive, and specific information to recognize the signs of renal dysfunction and how it contributes to identifying glomerular disease and renal transplant (RTx) decline has recently led to new investigations. The research for the detection of novel markers in urine samples, for the description of renal damage, is of high interest. In this regard, the evaluation of urinary cell mRNA expression has proved to be a common and simple method to obtain information on fine gene transcriptional changes leading to renal disease [11,12,13]. We have evaluated the urinary expression of a panel of specific podocyte genes, addressed to patients affected by renal disease with a clinical indication to RTx or native (N) renal biopsy (RBx)

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