title: Shroom3 contributes to the maintenance of the glomerular filtration barrier integrity.
abstract: Genome-wide association studies (GWAS) identify regions of the genome correlated with disease risk but are restricted in their ability to identify the underlying causative mechanism(s). Thus, GWAS are useful "roadmaps" that require functional analysis to establish the genetic and mechanistic structure of a particular locus. Unfortunately, direct functional testing in humans is limited, demonstrating the need for complementary approaches. Here we used an integrated approach combining zebrafish, rat, and human data to interrogate the function of an established GWAS locus (SHROOM3) lacking prior functional support for chronic kidney disease (CKD). Congenic mapping and sequence analysis in rats suggested Shroom3 was a strong positional candidate gene. Transferring a 6.1-Mb region containing the wild-type Shroom3 gene significantly improved the kidney glomerular function in FHH (fawn-hooded hypertensive) rat. The wild-type Shroom3 allele, but not the FHH Shroom3 allele, rescued glomerular defects induced by knockdown of endogenous shroom3 in zebrafish, suggesting that the FHH Shroom3 allele is defective and likely contributes to renal injury in the FHH rat. We also show for the first time that variants disrupting the actin-binding domain of SHROOM3 may cause podocyte effacement and impairment of the glomerular filtration barrier.
authors.affiliation: Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA; Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA;
authors.forename: Nan Cher
authors.initials: NC
authors.lastname: Yeo
authors.affiliation: Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA; Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA;
authors.forename: Caitlin C
authors.initials: CC
authors.lastname: O'Meara
authors.affiliation: Department of Molecular Medicine and Translational Science, Wake Forest School of Medicine, Winston-Salem, North Carolina 27157, USA; Center for Genomics and Personalized Medicine Research, Wake Forest School of Medicine, Winston-Salem, North Carolina 27157, USA;
authors.forename: Jason A
authors.initials: JA
authors.lastname: Bonomo
authors.affiliation: Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA;
authors.forename: Kerry N
authors.initials: KN
authors.lastname: Veth
authors.affiliation: Nephrology Division, Massachusetts General Hospital, Charlestown, Massachusetts 02129, USA;
authors.forename: Ritu
authors.initials: R
authors.lastname: Tomar
authors.affiliation: Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA; Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA;
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Flister
authors.affiliation: Nephrology Division, Massachusetts General Hospital, Charlestown, Massachusetts 02129, USA; Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA;
authors.forename: Iain A
authors.initials: IA
authors.lastname: Drummond
authors.affiliation: Center for Genomics and Personalized Medicine Research, Wake Forest School of Medicine, Winston-Salem, North Carolina 27157, USA; Department of Biochemistry, Wake Forest School of Medicine, Winston-Salem, North Carolina 27157, USA;
authors.forename: Donald W
authors.initials: DW
authors.lastname: Bowden
authors.affiliation: Center for Genomics and Personalized Medicine Research, Wake Forest School of Medicine, Winston-Salem, North Carolina 27157, USA; Department of Internal Medicine - Nephrology, Wake Forest School of Medicine, Winston-Salem, North Carolina 27157, USA;
authors.forename: Barry I
authors.initials: BI
authors.lastname: Freedman
authors.affiliation: Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA; Department of Dermatology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA;
authors.forename: Jozef
authors.initials: J
authors.lastname: Lazar
authors.affiliation: Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA;
authors.forename: Brian A
authors.initials: BA
authors.lastname: Link
authors.affiliation: Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA; Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA; Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA jacob@mcw.edu.
authors.forename: Howard J
authors.initials: HJ
authors.lastname: Jacob
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D020296:Animals, Congenic; D030801:Animals, Genetically Modified; D003001:Cloning, Molecular; D005091:Exons; D005260:Female; D056426:Genetic Loci; D014644:Genetic Variation; D055106:Genome-Wide Association Study; D060730:Glomerular Filtration Barrier / Q000378:metabolism*; D006801:Humans; D007674:Kidney Diseases / Q000235:genetics; D008297:Male; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017422:Sequence Analysis, DNA; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D008840:Microfilament Proteins; D012333:RNA, Messenger; C000596976:SHROOM3 protein, human; C577864:Shroom3 protein, zebrafish; D029961:Zebrafish Proteins