Universität zu Köln

Koehler, Sybille, Kuczkowski, Alexander, Kuehne, Lucas, Juengst, Christian, Hoehne, Martin, Grahammer, Florian, Eddy, Sean ORCID: 0000-0001-8578-3443, Kretzler, Matthias, Beck, Bodo B., Hoehfeld, Joerg, Schermer, Bernhard, Benzing, Thomas, Brinkkoetter, Paul T. and Rinschen, Markus M. (2020). Proteome Analysis of Isolated Podocytes Reveals Stress Responses in Glomerular Sclerosis. J. Am. Soc. Nephrol., 31 (3). S. 544 - 560. WASHINGTON: AMER SOC NEPHROLOGY. ISSN 1533-3450

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Abstract

Background Understanding podocyte-specific responses to injury at a systems level is difficult because injury leads to podocyte loss or an increase of extracellular matrix, altering glomerular cellular composition. Finding a window into early podocyte injury might help identify molecular pathways involved in the podocyte stress response. Methods We developed an approach to apply proteome analysis to very small samples of purified podocyte fractions. To examine podocytes in early disease states in FSGS mouse models, we used podocyte fractions isolated from individual mice after chemical induction of glomerular disease (with Doxorubicin or LPS). We also applied single-glomerular proteome analysis to tissue from patients with FSGS. Results Transcriptome and proteome analysis of glomeruli from patients with FSGS revealed an under-representation of podocyte-specific genes and proteins in late-stage disease. Proteome analysis of purified podocyte fractions from FSGS mouse models showed an early stress response that includes perturbations of metabolic, mechanical, and proteostasis proteins. Additional analysis revealed a high correlation between the amount of proteinuria and expression levels of the mechanosensor protein Filamin-B. Increased expression of Filamin-B in podocytes in biopsy samples from patients with FSGS, in single glomeruli from proteinuric rats, and in podocytes undergoing mechanical stress suggests that this protein has a role in detrimental stress responses. In Drosophila, nephrocytes with reduced filamin homo-log Cher displayed altered filtration capacity, but exhibited no change in slit diaphragm structure. Conclusions We identified conserved mechanisms of the podocyte stress response through ultrasensitive proteome analysis of human glomerular FSGS tissue and purified native mouse podocytes during early disease stages. This approach enables systematic comparisons of large-scale proteomics data and phenotypeto-protein correlation.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Koehler, Sybille UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuczkowski, Alexander UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuehne, Lucas UNSPECIFIED UNSPECIFIED UNSPECIFIED Juengst, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Hoehne, Martin UNSPECIFIED UNSPECIFIED UNSPECIFIED Grahammer, Florian UNSPECIFIED UNSPECIFIED UNSPECIFIED Eddy, Sean UNSPECIFIED orcid.org/0000-0001-8578-3443 UNSPECIFIED Kretzler, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Beck, Bodo B. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hoehfeld, Joerg UNSPECIFIED UNSPECIFIED UNSPECIFIED Schermer, Bernhard UNSPECIFIED UNSPECIFIED UNSPECIFIED Benzing, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Brinkkoetter, Paul T. UNSPECIFIED UNSPECIFIED UNSPECIFIED Rinschen, Markus M. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-343160
DOI:	10.1681/ASN.2019030312
Journal or Publication Title:	J. Am. Soc. Nephrol.
Volume:	31
Number:	3
Page Range:	S. 544 - 560
Date:	2020
Publisher:	AMER SOC NEPHROLOGY
Place of Publication:	WASHINGTON
ISSN:	1533-3450
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language GENE-EXPRESSION; CRE RECOMBINASE; KIDNEY-DISEASE; CELL; SYSTEM; QUANTIFICATION; BIOLOGY; MODEL Multiple languages Urology & Nephrology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/34316