Abstract
Nephrotic syndrome, a malfunction of the kidney glomerular filter, leads to proteinuria, edema and, in steroid-resistant nephrotic syndrome, end-stage kidney disease. Using positional cloning, we identified mutations in the phospholipase C epsilon gene (PLCE1) as causing early-onset nephrotic syndrome with end-stage kidney disease. Kidney histology of affected individuals showed diffuse mesangial sclerosis (DMS). Using immunofluorescence, we found PLCε1 expression in developing and mature glomerular podocytes and showed that DMS represents an arrest of normal glomerular development. We identified IQ motif–containing GTPase-activating protein 1 as a new interaction partner of PLCε1. Two siblings with a missense mutation in an exon encoding the PLCε1 catalytic domain showed histology characteristic of focal segmental glomerulosclerosis. Notably, two other affected individuals responded to therapy, making this the first report of a molecular cause of nephrotic syndrome that may resolve after therapy. These findings, together with the zebrafish model of human nephrotic syndrome generated by plce1 knockdown, open new inroads into pathophysiology and treatment mechanisms of nephrotic syndrome.
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Acknowledgements
We thank the affected individuals and their families for participation. We acknowledge R.H. Lyons for large-scale sequencing. We thank S.J. Allen and M. Petry for technical assistance and M. McKee for electron microscopy in zebrafish. GFP-tagged IQGAP1 constructs were provided by G. Bloom (University of Virginia). This research was supported by grants from the US National Institutes of Health to F.H., R.C.W. and L.B.H. (P50-DK039255), to R.C.W. (DK46073), to A.V.S. (R01-GM053536) to I.D. (R01-DK53093) and to G.G.K. (R01-DK56294) and by a grant from the KMD Foundation and the Thrasher Research Fund to F.H.; F.H. is the Frederick G.L. Huetwell Professor and a Doris Duke Distinguished Clinical Scientist. The work was further supported by the German Federal Ministry of Science and Education through the National Genome Research Network (C.B., H.C.H., G.N., P.N. and D.S.), by a EuReGene grant to D.M. (E.U., FP6005085) and by grants from the German Research Foundation (A.K., A.D. and T.G.).
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Supplementary information
Supplementary Fig. 1
Protein domains of human SRN3 (AB040949) as predicted by PFAM. (PDF 33 kb)
Supplementary Fig. 2
Characterization of two different anti-PLCε1 polyclonal antibodies. (PDF 74 kb)
Supplementary Fig. 3
PLCE1 mutation leads to renal histology of diffuse mesangial sclerosis (DMS) and is associated in with interrupted glomerular development. (PDF 691 kb)
Supplementary Fig. 4
CLUSTAL_W amino acid multiple sequence alignment of PLCε1 throughout evolution. (PDF 80 kb)
Supplementary Table 1
Exon-flanking and morpholino oligonucleotide primers used in human and zebrafish PLCE1 studies. (PDF 54 kb)
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Hinkes, B., Wiggins, R., Gbadegesin, R. et al. Positional cloning uncovers mutations in PLCE1 responsible for a nephrotic syndrome variant that may be reversible. Nat Genet 38, 1397–1405 (2006). https://doi.org/10.1038/ng1918
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DOI: https://doi.org/10.1038/ng1918
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