title: 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/Tns2ΔCcompound heterozygotes and Tns2ΔC/Tns2ΔChomozygous 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.
authors.affiliation: Laboratory of Laboratory Animal Science and Medicine, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan.
authors.forename: Kiyoma
authors.initials: K
authors.lastname: Marusugi
authors.forename: Kenta
authors.initials: K
authors.lastname: Nakano
authors.forename: Hayato
authors.initials: H
authors.lastname: Sasaki
authors.forename: Junpei
authors.initials: J
authors.lastname: Kimura
authors.forename: Rieko
authors.initials: R
authors.lastname: Yanobu-Takanashi
authors.forename: Tadashi
authors.initials: T
authors.lastname: Okamura
authors.forename: Nobuya
authors.initials: N
authors.lastname: Sasaki
mesh_terms: D000818:Animals; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004195:Disease Models, Animal; D005260:Female; D007678:Kidney Glomerulus / Q000473:pathology; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D050199:Podocytes / Q000378:metabolism; D038941:Polypyrimidine Tract-Binding Protein; D011507:Proteinuria / Q000209:etiology* / Q000235:genetics; D012598:Sclerosis; D000070876:Tensins / Q000235:genetics / Q000502:physiology*
chemical_list: D000070876:Tensins; D038941:Polypyrimidine Tract-Binding Protein; C511707:Tns2 protein, mouse