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pdx1 Knockout Leads to a Diabetic Nephropathy-Like Phenotype in Zebrafish and Identifies Phosphatidylethanolamine as Metabolite Promoting Early Diabetic Kidney Damage

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posted on 31.01.2022, 21:51 by Lucas M. Wiggenhauser, Lena Metzger, Katrin Bennewitz, Silas Soleymani, Mike Boger, Christoph T. Tabler, Ingrid Hausser, Carsten Sticht, Paulus Wohlfart, Nadine Volk, Elena Heidenreich, Michael Buettner, Hans-Peter Hammes, Jens Kroll
The pdx1-/- zebrafish mutant was recently established as a novel animal model of diabetic retinopathy. Here, we investigate whether knockout of pdx1 also leads to diabetic kidney disease (DKD). pdx1-/- larvae exhibit several signs of early DKD such as glomerular hypertrophy, impairments in the filtration barrier corresponding to microalbuminuria and glomerular basement membrane (GBM) thickening. Adult pdx1-/- mutants show progressive GBM thickening in comparison to the larval state. Heterozygous pdx1 knockout also leads to glomerular hypertrophy as initial establishment of DKD similar to the pdx1-/- larvae. RNA sequencing (RNA-seq) of adult pdx1+/- kidneys uncovered regulations in multiple expected diabetic pathways related to podocyte disruption and hinting at early vascular dysregulation without obvious morphological alterations. Metabolome analysis and pharmacological intervention experiments revealed the contribution of phosphatidylethanolamine (PtdE) in the early establishment of kidney damage. In conclusion, this study identified the pdx1 mutant as a novel model for the study of DKD showing signs of the early disease progression already in larval stage and several selective features of later DKD in adult mutants.

Funding

The study was supported by grants from Deutsche Forschungsgemeinschaft (CRC 1118 and IRTG 1874/2 DIAMICOM). We thank Gernot Poschet and Ruediger Hell from the Metabolomics Core Technology Platform of the Excellence cluster “CellNetworks” (University of Heidelberg), and the Deutsche Forschungsgemeinschaft (grant ZUK 40/2010-3009262) for support with UPLC and GC/MS-based metabolite quantification. We thank Elke Deckert and Uwe Schwahn (Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany) for technical and analytical support regarding the RNA sequencing, and the contributions of Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany within the collaboration IRTG 1874/2 DIAMICOM.

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