Functional validation of tensin2 SH2-PTB domain by CRISPR/Cas9-mediated genome editing

Abstract

Podocytes are terminally differentiated and highly specialized cells in the glomerulus, and they form a crucial component of the glomerular filtration barrier. The ICGN mouse is a model of glomerular dysfunction that shows gross morphological changes in the podocyte foot process, accompanied by proteinuria. Previously, we demonstrated that proteinuria in ICR-derived glomerulonephritis mouse ICGN mice might be caused by a deletion mutation in the tensin2 (Tns2) gene (designated Tns2nph). To test whether this mutation causes the mutant phenotype, we created knockout (KO) mice carrying a Tns2 protein deletion in the C-terminal Src homology and phosphotyrosine binding (SH2-PTB) domains (designated Tns2ΔC) via CRISPR/Cas9-mediated genome editing. Tns2nph/Tns2ΔC compound heterozygotes and Tns2ΔC/Tns2ΔC homozygous KO mice displayed podocyte abnormalities and massive proteinuria similar to ICGN mice, indicating that these two mutations are allelic. Further, this result suggests that the SH2-PTB domain of Tns2 is required for podocyte integrity. Tns2 knockdown in a mouse podocyte cell line significantly enhanced actin stress fiber formation and cell migration. Thus, this study provides evidence that alteration of actin remodeling resulting from Tns2 deficiency causes morphological changes in podocytes and subsequent proteinuria.

AMPK signalling: Implications for podocyte biology in diabetic nephropathy

“…Tensin that contains integrin binding region is known as cytoskeleton linker protein and signal transducer [Lo et al., ]. Growing body of evidence has shown that tensin2 is a relevant for the maintenance of normal kidney function [Lo et al., ; Cho et al., ] Moreover, tensin mutation or deficiency in mice have been indicated to alterate of actin remodelling in podocytes, which results in foot process effacement and massive albuminuria [Marusugi et al., ; Takahashi et al., ]. Tensins are close relatives of phosphatase and tensin homolog (PTEN).…”

Novel blood pressure locus and gene discovery using GWAS and expression datasets from blood and the kidney

“…SNP rs73099903 was in strong linkage disequilibrium (LD r 2 >0.9) with the SNP most strongly associated with TNS2 expression in the BIOS resource. TNS2 encodes a tensin focal adhesion molecule and may have a role in renal function (26).…”

C1-Ten is a PTPase of nephrin, regulating podocyte hypertrophy through mTORC1 activation

“…The ICGN mouse, an animal model for nephrotic syndrome, shows severe proteinuria and glomerulosclerosis with impairment of nephrin and synaptopodin 35 . Also, podocyte abnormalities and proteinuria are observed in mice with C1-Ten SH2-PTB domain deletion mutant 36 . These results and our study suggest that both C1-Ten WT overexpression and C1-Ten mutation contribute to podocyte dysfunction and proteinuria.…”