Integration of Cistromic and Transcriptomic Analyses Identifies Nphs2, Mafb, and Magi2 as Wilms' Tumor 1 Target Genes in Podocyte Differentiation and Maintenance

J Am Soc Nephrol. 2015 Sep;26(9):2118-28. doi: 10.1681/ASN.2014080819. Epub 2015 Jan 2.

Abstract

The Wilms' tumor suppressor gene 1 (WT1) encodes a zinc finger transcription factor. Mutation of WT1 in humans leads to Wilms' tumor, a pediatric kidney tumor, or other kidney diseases, such as Denys-Drash and Frasier syndromes. We showed previously that inactivation of WT1 in podocytes of adult mice results in proteinuria, foot process effacement, and glomerulosclerosis. However, the WT1-dependent transcriptional network regulating podocyte development and maintenance in vivo remains unknown. Here, we performed chromatin immunoprecipitation followed by high-throughput sequencing with glomeruli from wild-type mice. Additionally, we performed a cDNA microarray screen on an inducible podocyte-specific WT1 knockout mouse model. By integration of cistromic and transcriptomic analyses, we identified the WT1 targetome in mature podocytes. To further analyze the function and targets of WT1 in podocyte maturation, we used an Nphs2-Cre model, in which WT1 is deleted during podocyte differentiation. These mice display anuria and kidney hemorrhage and die within 24 hours after birth. To address the evolutionary conservation of WT1 targets, we performed functional assays using zebrafish as a model and identified Nphs2, Mafb, and Magi2 as novel WT1 target genes required for podocyte development. Our data also show that both Mafb and Magi2 are required for normal development of the embryonic zebrafish kidney. Collectively, our work provides insights into the transcriptional networks controlled by WT1 and identifies novel WT1 target genes that mediate the function of WT1 in podocyte differentiation and maintenance.

Keywords: ESRD; chronic hemodialysis; economic analysis; epidemiology and outcomes; outcomes; vascular access.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Animals
  • Cell Differentiation / genetics*
  • Gene Expression Profiling
  • Gene Expression Regulation*
  • Genes, Wilms Tumor / physiology*
  • Guanylate Kinases / genetics
  • Intracellular Signaling Peptides and Proteins / genetics
  • MafB Transcription Factor / genetics
  • Membrane Proteins / genetics
  • Mice
  • Mice, Knockout
  • Nerve Tissue Proteins / genetics
  • Oligonucleotide Array Sequence Analysis
  • Podocytes / physiology*
  • Repressor Proteins / genetics*
  • WT1 Proteins / genetics*
  • Zebrafish
  • Zebrafish Proteins / genetics*

Substances

  • Adaptor Proteins, Signal Transducing
  • Intracellular Signaling Peptides and Proteins
  • MafB Transcription Factor
  • Mafb protein, mouse
  • Membrane Proteins
  • NPHS2 protein
  • Nerve Tissue Proteins
  • Repressor Proteins
  • WT1 Proteins
  • WT1 protein, mouse
  • Wt1a protein, zebrafish
  • Zebrafish Proteins
  • mafb protein, zebrafish
  • magi2a protein, zebrafish
  • Guanylate Kinases
  • Magi2 protein, mouse