title: TRPC6 G757D Loss-of-Function Mutation Associates with FSGS.
abstract: FSGS is a CKD with heavy proteinuria that eventually progresses to ESRD. Hereditary forms of FSGS have been linked to mutations in the transient receptor potential cation channel, subfamily C, member 6 (TRPC6) gene encoding a nonselective cation channel. Most of these TRPC6 mutations cause a gain-of-function phenotype, leading to calcium-triggered podocyte cell death, but the underlying molecular mechanisms are unclear. We studied the molecular effect of disease-related mutations using tridimensional in silico modeling of tetrameric TRPC6. Our results indicated that G757 is localized in a domain forming a TRPC6-TRPC6 interface and predicted that the amino acid exchange G757D causes local steric hindrance and disruption of the channel complex. Notably, functional characterization of model interface domain mutants suggested a loss-of-function phenotype. We then characterized 19 human FSGS-related TRPC6 mutations, the majority of which caused gain-of-function mutations. However, five mutations (N125S, L395A, G757D, L780P, and R895L) caused a loss-of-function phenotype. Coexpression of wild-type TRPC6 and TRPC6 G757D, mimicking heterozygosity observed in patients, revealed a dominant negative effect of TRPC6 G757D. Our comprehensive analysis of human disease-causing TRPC6 mutations reveals loss of TRPC6 function as an additional concept of hereditary FSGS and provides molecular insights into the mechanism responsible for the loss-of-function phenotype of TRPC6 G757D in humans.
authors.affiliation: Department of Pharmacology and Experimental Therapy, Institute of Experimental and Clinical Pharmacology and Toxicology, Eberhard Karls University Hospitals and Clinics and Interfaculty Center of Pharmacogenomics and Drug Research, University of Tübingen, Tübingen, Germany;
authors.forename: Marc
authors.initials: M
authors.lastname: Riehle
authors.affiliation: Pediatric Nephrology, Pediatrics II, University of Duisburg-Essen, Essen, Germany;
authors.forename: Anja K
authors.initials: AK
authors.lastname: Büscher
authors.affiliation: German Cancer Consortium, Heidelberg, Germany; Charité University Medicine Berlin, Structural Bioinformatics Group, Institute of Physiology and Experimental Clinical Research Center, Berlin, Germany;
authors.forename: Björn-Oliver
authors.initials: BO
authors.lastname: Gohlke
authors.affiliation: Nephrology/Intensive Care, Experimental and Clinical Research Center and Max Delbrück Center for Molecular Medicine, Charité University Medicine Berlin, Berlin, Germany;
authors.forename: Mario
authors.initials: M
authors.lastname: Kaßmann
authors.affiliation: Nephro-Urology Unit, University College London Institute of Child Health, London, United Kingdom;
authors.forename: Maria
authors.initials: M
authors.lastname: Kolatsi-Joannou
authors.affiliation: Institute of Pathology, University Hospital of Hannover, Hannover, Germany;
authors.forename: Jan H
authors.initials: JH
authors.lastname: Bräsen
authors.affiliation: Center of Nephrology and Metabolism, Weisswasser, Germany;
authors.forename: Mato
authors.initials: M
authors.lastname: Nagel
authors.affiliation: Institute of Pathology, University Hospital of Cologne, Cologne, Germany; and.
authors.forename: Jan U
authors.initials: JU
authors.lastname: Becker
authors.affiliation: Nephro-Urology Unit, University College London Institute of Child Health, London, United Kingdom;
authors.forename: Paul
authors.initials: P
authors.lastname: Winyard
authors.affiliation: Pediatric Nephrology, Pediatrics II, University of Duisburg-Essen, Essen, Germany;
authors.forename: Peter F
authors.initials: PF
authors.lastname: Hoyer
authors.affiliation: German Cancer Consortium, Heidelberg, Germany; Charité University Medicine Berlin, Structural Bioinformatics Group, Institute of Physiology and Experimental Clinical Research Center, Berlin, Germany;
authors.forename: Robert
authors.initials: R
authors.lastname: Preissner
authors.affiliation: Deutsche Forschungsanstalt für Lebensmittelchemie, Molekulare Zellphysiologie und Chemorezeption, Freising, Germany.
authors.forename: Dietmar
authors.initials: D
authors.lastname: Krautwurst
authors.affiliation: Nephrology/Intensive Care, Experimental and Clinical Research Center and Max Delbrück Center for Molecular Medicine, Charité University Medicine Berlin, Berlin, Germany;
authors.forename: Maik
authors.initials: M
authors.lastname: Gollasch
authors.affiliation: Pediatric Nephrology, Pediatrics II, University of Duisburg-Essen, Essen, Germany; stefanie.weber@uk-essen.de christian.harteneck@uni-tuebingen.de.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Weber
authors.affiliation: Department of Pharmacology and Experimental Therapy, Institute of Experimental and Clinical Pharmacology and Toxicology, Eberhard Karls University Hospitals and Clinics and Interfaculty Center of Pharmacogenomics and Drug Research, University of Tübingen, Tübingen, Germany; stefanie.weber@uk-essen.de christian.harteneck@uni-tuebingen.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Harteneck
mesh_terms: D004252:DNA Mutational Analysis; D005923:Glomerulosclerosis, Focal Segmental / Q000235:genetics* / Q000503:physiopathology; D006801:Humans; D009154:Mutation; D050052:TRPC Cation Channels / Q000235:genetics*; D000074287:TRPC6 Cation Channel
chemical_list: D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C117032:TRPC6 protein, human
keywords: calcium; glomerular disease; glomerulosclerosis; ion channel; kidney disease