title: Mucin-1 Increases Renal TRPV5 Activity In Vitro, and Urinary Level Associates with Calcium Nephrolithiasis in Patients.
abstract: Hypercalciuria is a major risk factor for nephrolithiasis. We previously reported that Uromodulin (UMOD) protects against nephrolithiasis by upregulating the renal calcium channel TRPV5. This channel is crucial for calcium reabsorption in the distal convoluted tubule (DCT). Recently, mutations in the gene encoding Mucin-1 (MUC1) were found to cause autosomal dominant tubulointerstitial kidney disease, the same disease caused by UMOD mutations. Because of the similarities between UMOD and MUC1 regarding associated disease phenotype, protein structure, and function as a cellular barrier, we examined whether urinary MUC1 also enhances TRPV5 channel activity and protects against nephrolithiasis. We established a semiquantitative assay for detecting MUC1 in human urine and found that, compared with controls (n=12), patients (n=12) with hypercalciuric nephrolithiasis had significantly decreased levels of urinary MUC1. Immunofluorescence showed MUC1 in the thick ascending limb, DCT, and collecting duct. Applying whole-cell patch-clamp recording of HEK cells, we found that wild-type but not disease mutant MUC1 increased TRPV5 activity by impairing dynamin-2- and caveolin-1-mediated endocytosis of TRPV5. Coimmunoprecipitation confirmed a physical interaction between TRPV5 and MUC1. However, MUC1 did not increase the activity of N-glycan-deficient TRPV5. MUC1 is characterized by variable number tandem repeats (VNTRs) that bind the lectin galectin-3; galectin-3 siRNA but not galectin-1 siRNA prevented MUC1-induced upregulation of TRPV5 activity. Additionally, MUC1 lacking VNTRs did not increase TRPV5 activity. Our results suggest that MUC1 forms a lattice with the N-glycan of TRPV5 via galectin-3, which impairs TRPV5 endocytosis and increases urinary calcium reabsorption.
authors.affiliation: Pediatric Nephrology.
authors.forename: Mingzhu
authors.initials: M
authors.lastname: Nie
authors.affiliation: Pediatric Nephrology.
authors.forename: Manjot S
authors.initials: MS
authors.lastname: Bal
authors.affiliation: Nephrology, and.
authors.forename: Zhufeng
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Pediatric Nephrology.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.affiliation: Pediatric Nephrology.
authors.forename: Carolina
authors.initials: C
authors.lastname: Rivera
authors.affiliation: Institute of Human Genetics, University of Cologne, Cologne, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Wenzel
authors.affiliation: Institute of Human Genetics, University of Cologne, Cologne, Germany.
authors.forename: Bodo B
authors.initials: BB
authors.lastname: Beck
authors.affiliation: Charles and Jane Pak Center for Mineral Metabolism and Clinical Research, University of Texas Southwestern Medical Center, Dallas, Texas; and.
authors.forename: Khashayar
authors.initials: K
authors.lastname: Sakhaee
authors.affiliation: Nephrology, and.
authors.forename: Denise K
authors.initials: DK
authors.lastname: Marciano
authors.affiliation: Pediatric Nephrology, matthias.wolf@utsouthwestern.edu.
authors.forename: Matthias T F
authors.initials: MT
authors.lastname: Wolf
mesh_terms: D002118:Calcium / Q000032:analysis; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D018396:Mucin-1 / Q000502:physiology* / Q000652:urine*; D053040:Nephrolithiasis / Q000209:etiology* / Q000652:urine*; D050916:TRPV Cation Channels / Q000502:physiology*; D015854:Up-Regulation
chemical_list: D018396:Mucin-1; D050916:TRPV Cation Channels; C492239:TRPV5 protein, human; D002118:Calcium
keywords: calcium; electrophysiology; endocytosis; hypercalciuria; ion channel; kidney stones