Proteome Analysis of Isolated Podocytes Reveals Stress Responses in Glomerular Sclerosis.
glomerular disease
podocyte
renal injury
Journal
Journal of the American Society of Nephrology : JASN
ISSN: 1533-3450
Titre abrégé: J Am Soc Nephrol
Pays: United States
ID NLM: 9013836
Informations de publication
Date de publication:
03 2020
03 2020
Historique:
received:
28
03
2019
accepted:
04
12
2019
pubmed:
13
2
2020
medline:
11
11
2020
entrez:
13
2
2020
Statut:
ppublish
Résumé
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. 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. Transcriptome and proteome analysis of glomeruli from patients with FSGS revealed an underrepresentation 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 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 phenotype-to-protein correlation.
Sections du résumé
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 underrepresentation 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
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 phenotype-to-protein correlation.
Identifiants
pubmed: 32047005
pii: ASN.2019030312
doi: 10.1681/ASN.2019030312
pmc: PMC7062218
doi:
Substances chimiques
FLNB protein, mouse
0
Filamins
0
Types de publication
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
544-559Subventions
Organisme : NIDDK NIH HHS
ID : P30 DK081943
Pays : United States
Informations de copyright
Copyright © 2020 by the American Society of Nephrology.
Références
Sci Signal. 2014 Feb 04;7(311):ra12
pubmed: 24497609
J Cell Sci. 2018 Apr 13;131(8):
pubmed: 29654161
Cell Death Differ. 2016 May;23(5):776-86
pubmed: 26586575
Kidney Int. 2017 Oct;92(4):909-921
pubmed: 28554737
Kidney Int. 2018 Jan;93(1):110-127
pubmed: 28754552
Sci Signal. 2017 Apr 11;10(474):
pubmed: 28400537
F1000Res. 2017 Feb 9;6:121
pubmed: 28232870
J Am Soc Nephrol. 2016 Dec;27(12):3600-3610
pubmed: 27026370
Nature. 2011 May 19;473(7347):337-42
pubmed: 21593866
Clin J Am Soc Nephrol. 2017 Mar 7;12(3):502-517
pubmed: 28242845
Curr Biol. 2013 Mar 4;23(5):430-5
pubmed: 23434281
Mol Biol Cell. 2011 Jun 15;22(12):2010-30
pubmed: 21508316
Proteomics Clin Appl. 2015 Dec;9(11-12):1053-68
pubmed: 25907645
J Am Soc Nephrol. 2015 Sep;26(9):2097-104
pubmed: 25636411
J Cell Sci. 2017 Sep 1;130(17):2781-2788
pubmed: 28808089
Diabetes. 2009 Feb;58(2):469-77
pubmed: 19017763
Nat Commun. 2019 Sep 11;10(1):4130
pubmed: 31511532
J Am Soc Nephrol. 2017 Sep;28(9):2641-2653
pubmed: 28424277
Nat Rev Nephrol. 2019 Jul;15(7):393-411
pubmed: 31036905
J Am Soc Nephrol. 2002 Dec;13(12):3005-15
pubmed: 12444221
J Am Soc Nephrol. 2014 Jul;25(7):1509-22
pubmed: 24511133
Genome Res. 2013 Nov;23(11):1862-73
pubmed: 23950145
Kidney Int. 2017 Jun;91(6):1510-1517
pubmed: 28187984
Cell Rep. 2018 May 22;23(8):2495-2508
pubmed: 29791858
Proteomics. 2009 Oct;9(19):4519-28
pubmed: 19688724
Nat Biotechnol. 2008 Dec;26(12):1367-72
pubmed: 19029910
Nat Med. 2017 Apr;23(4):429-438
pubmed: 28218918
Nat Med. 2017 Jan;23(1):100-106
pubmed: 27941791
J Am Soc Nephrol. 2017 Oct;28(10):2867-2878
pubmed: 28724775
Genesis. 2007 Sep;45(9):593-605
pubmed: 17868096
Am J Pathol. 2010 Oct;177(4):1674-86
pubmed: 20847290
Nature. 2009 Jan 15;457(7227):322-6
pubmed: 18971929
BMC Bioinformatics. 2012;13 Suppl 16:S12
pubmed: 23176165
Genesis. 2003 Jan;35(1):39-42
pubmed: 12481297
PLoS One. 2011;6(12):e28710
pubmed: 22194892
Kidney Int. 2013 Jun;83(6):1052-64
pubmed: 23364521
J Cell Biol. 2019 Aug 5;218(8):2481-2491
pubmed: 31315944
Hum Mol Genet. 2017 Feb 15;26(4):768-780
pubmed: 28164240
Bioinformatics. 2013 Jul 15;29(14):1830-1
pubmed: 23740750
Nucleic Acids Res. 2019 Jan 8;47(D1):D442-D450
pubmed: 30395289
Bioinformatics. 2010 Apr 1;26(7):966-8
pubmed: 20147306
J Am Soc Nephrol. 2013 Feb;24(2):191-7
pubmed: 23291470
Nat Commun. 2014 Aug 14;5:4656
pubmed: 25120197
Mol Cell Proteomics. 2014 Sep;13(9):2513-26
pubmed: 24942700
Physiol Rev. 2003 Jan;83(1):253-307
pubmed: 12506131
Hum Mol Genet. 2016 Apr 1;25(7):1328-44
pubmed: 26792178
Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5116-21
pubmed: 11309499
Front Pediatr. 2017 Dec 07;5:262
pubmed: 29270398
Mol Syst Biol. 2014 Oct 30;10:757
pubmed: 25358341
Physiol Rev. 2018 Oct 1;98(4):2571-2606
pubmed: 30182799
J Am Soc Nephrol. 2017 May;28(5):1521-1533
pubmed: 27932481
Kidney Int. 2018 Jun;93(6):1308-1319
pubmed: 29530281
Kidney Int. 2018 Oct;94(4):795-808
pubmed: 30093081
Cell. 2009 Mar 6;136(5):913-25
pubmed: 19269368