title: Wolf-Hirschhorn syndrome candidate 1-like 1 epigenetically regulates nephrin gene expression.
abstract: Altered expression of nephrin underlies the pathophysiology of proteinuria in both congenital and acquired nephrotic syndrome. However, the epigenetic mechanisms of nephrin gene regulation remain elusive. Here, we show that Wolf-Hirschhorn syndrome candidate 1-like 1 long form (WHSC1L1-L) is a novel epigenetic modifier of nephrin gene regulation. WHSC1L1-L was associated with histone H3K4 and H3K36 in human embryonic kidney cells. WHSC1L1-L gene was expressed in the podocytes, and functional protein product was detected in these cells. WHSC1L1-L was found to bind nephrin but not other podocyte-specific gene promoters, leading to its inhibition/suppression, abrogating the stimulatory effect of WT1 and NF-κB. Gene knockdown of WHSC1L1-L in primary cultured podocytes accelerated the transcription of nephrin but not CD2AP. An in vivo zebrafish study involving the injection of Whsc1l1 mRNA into embryos demonstrated an apparent reduction of nephrin mRNA but not podocin and CD2AP mRNA. Immunohistochemistry showed that both WHSC1L1-L and nephrin emerged at the S-shaped body stage in glomeruli. Immunofluorescence and confocal microscopy displayed WHSC1L1 to colocalize with trimethylated H3K4 in the glomerular podocytes. Chromatin immunoprecipitation assay revealed the reduction of the association of trimethylated H3K4 at the nephrin promoter regions. Finally, nephrin mRNA was upregulated in the glomerulus at the early proteinuric stage of mouse nephrosis, which was associated with the reduction of WHSC1L1. In conclusion, our results demonstrate that WHSC1L1-L acts as a histone methyltransferase in podocytes and regulates nephrin gene expression, which may in turn contribute to the integrity of the slit diaphragm of the glomerular filtration barrier.
authors.affiliation: Department of Pediatrics, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
authors.forename: Yugo
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Department of Medical Biochemistry and Biophysics, Division of Matrix Biology, Karolinska Institute, Stockholm, Sweden.
authors.forename: Kan
authors.initials: K
authors.lastname: Katayama
authors.affiliation: Department of Pediatrics, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
authors.forename: Yukino
authors.initials: Y
authors.lastname: Nishibori
authors.affiliation: Department of Anatomy, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Akimoto
authors.affiliation: Department of Anatomy, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kudo
authors.affiliation: Department of Pediatrics, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Kurayama
authors.affiliation: Department of Pediatrics, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Hada
authors.affiliation: Department of Pediatrics, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
authors.forename: Shohei
authors.initials: S
authors.lastname: Takahashi
authors.affiliation: Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Mitaka, Tokyo, Japan; and.
authors.forename: Toru
authors.initials: T
authors.lastname: Kimura
authors.affiliation: Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Mitaka, Tokyo, Japan; and.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Fukutomi
authors.affiliation: Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Mitaka, Tokyo, Japan; and.
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Katada
authors.affiliation: Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Mitaka, Tokyo, Japan; and.
authors.forename: Junichi
authors.initials: J
authors.lastname: Suehiro
authors.affiliation: Molecular Medicine Center and Department of Medical Chemistry and Biochemistry, Medical University of Sofia, Sofia, Bulgaria.
authors.forename: Olga
authors.initials: O
authors.lastname: Beltcheva
authors.affiliation: Department of Medical Biochemistry and Biophysics, Division of Matrix Biology, Karolinska Institute, Stockholm, Sweden.
authors.forename: Karl
authors.initials: K
authors.lastname: Tryggvason
authors.affiliation: Department of Pediatrics, Kyorin University School of Medicine, Mitaka, Tokyo, Japan; kuniyan@ks.kyorin-u.ac.jp.
authors.forename: Kunimasa
authors.initials: K
authors.lastname: Yan
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D004317:Doxorubicin; D044127:Epigenesis, Genetic; D005786:Gene Expression Regulation; D015971:Gene Expression Regulation, Enzymologic; D057809:HEK293 Cells; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics* / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008745:Methylation; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009404:Nephrotic Syndrome / Q000139:chemically induced / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D050199:Podocytes / Q000201:enzymology* / Q000473:pathology; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D014162:Transfection; D015027:Zebrafish / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C114545:CD2-associated protein; D003598:Cytoskeletal Proteins; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C407223:NPHS2 protein; D009687:Nuclear Proteins; D029961:Zebrafish Proteins; C113267:nephrin; D004317:Doxorubicin; D011495:Histone-Lysine N-Methyltransferase; C000621748:WHSC1L1 protein, mouse
keywords: Nephrin; epigenetics; gene regulation: development; podocyte