Revisiting the gene mutations and protein profile of WT 9‐12: An autosomal dominant polycystic kidney disease cell line

Abstract

AbstractWT 9‐12 is one of the cell lines commonly used for autosomal dominant polycystic kidney disease (ADPKD) studies. Previous studies had described the PKD gene mutations and polycystin expression in WT 9‐12. Nonetheless, the mutations occurring in other ADPKD‐associated genes have not been investigated. This study aims to revisit these mutations and protein profile of WT 9‐12. Whole genome sequencing verified the presence of truncation mutation at amino acid 2556 (Q2556X) in PKD1 gene of WT 9‐12. Besides, those variations with high impacts included single nucleotide polymorphisms (rs8054182, rs117006360, and rs12925771) and insertions and deletions (InDels) (rs145602984 and rs55980345) in PKD1L2; InDel (rs1296698195) in PKD1L3; and copy number variations in GANAB. Protein profiles generated from the total proteins of WT 9‐12 and HK‐2 cells were compared using isobaric tags for relative and absolute quantitation (iTRAQ) analysis. Polycystin‐1 was absent in WT 9‐12. The gene ontology enrichment and reactome pathway analyses revealed that the upregulated and downregulated proteins of WT 9‐12 relative to HK‐2 cell line leaded to signaling pathways related to immune response and amino acid metabolism, respectively. The ADPKD‐related mutations and signaling pathways associated with differentially expressed proteins in WT 9‐12 may help researchers in cell line selection for their studies.