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Am J Physiol Renal Physiol. 2019 Jan 1;316(1):F204-F213. doi: 10.1152/ajprenal.00470.2018. Epub 2018 Nov 7.

Identifying cystogenic paracrine signaling molecules in cyst fluid of patients with polycystic kidney disease.

Author information

1
Department of Internal Medicine, Division of Nephrology and Transplantation, Erasmus Medical Center, Rotterdam , The Netherlands.
2
Department of Cell Biology, Erasmus Medical Center, Rotterdam , The Netherlands.
3
Department of Developmental Biology, Erasmus Medical Center, Rotterdam , The Netherlands.
4
Department of Molecular Genetics, Erasmus Medical Center, Rotterdam , The Netherlands.
5
Department of Urology, Erasmus Medical Center, Rotterdam , The Netherlands.
6
Department of Pathology, Erasmus Medical Center, Rotterdam , The Netherlands.
7
Proteomics Center, Erasmus Medical Center, Rotterdam , The Netherlands.

Abstract

In autosomal dominant polycystic kidney disease (ADPKD) paracrine signaling molecules in cyst fluid can induce proliferation and cystogenesis of neighboring renal epithelial cells. However, the identity of this cyst-inducing factor is still unknown. The aim of this study was to identify paracrine signaling proteins in cyst fluid using a 3D in vitro cystogenesis assay. We collected cyst fluid from 15 ADPKD patients who underwent kidney or liver resection (55 cysts from 13 nephrectomies, 5 cysts from 2 liver resections). For each sample, the ability to induce proliferation and cyst formation was tested using the cystogenesis assay (RPTEC/TERT1 cells in Matrigel with cyst fluid added for 14 days). Kidney cyst fluid induced proliferation and cyst growth of renal epithelial cells in a dose-dependent fashion. Liver cyst fluid also induced cystogenesis. Using size exclusion chromatography, 56 cyst fluid fractions were obtained of which only the fractions between 30 and 100 kDa showed cystogenic potential. Mass spectrometry analysis of samples that tested positive or negative in the assay identified 43 candidate cystogenic proteins. Gene ontology analysis showed an enrichment for proteins classified as enzymes, immunity proteins, receptors, and signaling proteins. A number of these proteins have previously been implicated in ADPKD, including secreted frizzled-related protein 4, S100A8, osteopontin, and cysteine rich with EGF-like domains 1. In conclusion, both kidney and liver cyst fluids contain paracrine signaling molecules that drive cyst formation. Using size exclusion chromatography and mass spectrometry, we procured a candidate list for future studies. Ultimately, cystogenic paracrine signaling molecules may be targeted to abrogate cystogenesis in ADPKD.

KEYWORDS:

ADPKD; cyst fluid; kidney; liver; mass spectrometry; paracrine signaling

PMID:
30403162
DOI:
10.1152/ajprenal.00470.2018
[Indexed for MEDLINE]
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