Dataset Information

Analysis of podocytes from doxorubicin treated mice

ABSTRACT: Podocyte injury is a major cause of chronic kidney disease, but the involved signaling pathways are largely unknown. Here, we used ultrasensitive proteomics to investigate the response of native podocytes from mice with Doxorubicin-induced injury. We observed perturbations of known propagating pathways of glomerular disease such as Jak-Stat signaling and cell adhesion but also in metabolism and proteins responding to mechanical stress. Using a phenotype-guided approach, we prioritized candidates by aligning proteinuria in individual mice with corresponding podocyte protein expression data. The protein Tropomodulin was decreased and correlated negatively with urinary albumin, and Villin and the mechanosensor protein Filamin-B were increased and correlated positively. We employed Drosophila nephrocytes for further functional analysis of prioritized proteins and studied filtration function and endocytosis. Loss of Villin and Tropomodulin led to an impaired function and reduced expression of endocytosis machineries as evidenced by nephrocyte garland proteomes. Loss of Cher, the orthologue of Filamin-B, led to increased endocytosis in nephrocytes, and Filamin-B abundance in single glomeruli correlated with the amount of deposited albumin in the same glomerulus in proteinuric rats. Filamin-B expression could also be increased by applying mechanical stress to podocytes in vitro, but Filamin-B was not increased in podocyte proteomes in LPS-induced injury despite similar regulation of pathways on the global proteome level. In conclusion, this study identified conserved mechanisms of podocyte stress response employing functional proteomics in purified native mouse podocytes and Drosophila nephrocytes.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Glomerular Visceral Epithelial Cell, Kidney

SUBMITTER: Markus Rinschen

LAB HEAD: Markus Rinschen

PROVIDER: PXD012063 | Pride | 2020-07-29

REPOSITORIES: Pride

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			Action	DRS
	20171012_Q2_UC_ColID_415_ProjID_796_Benzing_Sample_1.raw	Raw
	20171012_Q2_UC_ColID_415_ProjID_796_Benzing_Sample_10.raw	Raw
	20171012_Q2_UC_ColID_415_ProjID_796_Benzing_Sample_11.raw	Raw
	20171012_Q2_UC_ColID_415_ProjID_796_Benzing_Sample_2.raw	Raw
	20171012_Q2_UC_ColID_415_ProjID_796_Benzing_Sample_3.raw	Raw

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Publications

Proteome Analysis of Isolated Podocytes Reveals Stress Responses in Glomerular Sclerosis.

Koehler Sybille S Kuczkowski Alexander A Kuehne Lucas L Jüngst Christian C Hoehne Martin M Grahammer Florian F Eddy Sean S Kretzler Matthias M Beck Bodo B BB Höhfeld Jörg J Schermer Bernhard B Benzing Thomas T Brinkkoetter Paul T PT Rinschen Markus M MM

Journal of the American Society of Nephrology : JASN 20200211 3

<h4>Background</h4>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.<h4>Methods</h4>We developed an approach to apply proteome analysis to very small samples of purified podocyte fractions. To examine podocytes in early d ...[more]

PMID: 32047005

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