We performed next generation sequencing in patients with familial steroid sensitive nephrotic syndrome (SSNS) and identified a homozygous segregating variant (p.H310Y) in the gene encoding clavesin-1 (CLVS1) in a consanguineous family with three affected individuals. Knockdown of the clavesin gene in zebrafish (clvs2) produced edema phenotypes due to disruption of podocyte structure and loss of glomerular filtration barrier integrity that can be rescued by WT CLVS1 but not the p.H310Y variant. Analysis of cultured human podocytes with CRISPR-Cas9 mediated CLVS1 knockout or homozygous H310Y knockin revealed deficits in clathrin-mediated endocytosis and increased susceptibility to apoptosis that could be rescued ... More
We performed next generation sequencing in patients with familial steroid sensitive nephrotic syndrome (SSNS) and identified a homozygous segregating variant (p.H310Y) in the gene encoding clavesin-1 (CLVS1) in a consanguineous family with three affected individuals. Knockdown of the clavesin gene in zebrafish (clvs2) produced edema phenotypes due to disruption of podocyte structure and loss of glomerular filtration barrier integrity that can be rescued by WT CLVS1 but not the p.H310Y variant. Analysis of cultured human podocytes with CRISPR-Cas9 mediated CLVS1 knockout or homozygous H310Y knockin revealed deficits in clathrin-mediated endocytosis and increased susceptibility to apoptosis that could be rescued with corticosteroid treatment, mimicking the steroid-responsiveness observed in SSNS patients. The p.H310Y variant also disrupts binding of clavesin-1 to alpha-tocopherol transfer protein, resulting in increased reactive oxygen species (ROS) accumulation in CLVS1-deficient podocytes. Treatment of CLVS1 knockout or homozygous H310Y knockin podocytes with pharmacological ROS inhibitors restored viability to control levels. Taken together, this data identifies CLVS1 as a candidate gene for SSNS, provides insight into therapeutic effects of corticosteroids on podocyte cellular dynamics and adds to the growing evidence on the importance of endocytosis and oxidative stress regulation to podocyte function.
