Detection of a splice site variant in a patient with glomerulopathy and fibronectin deposits

Yurika Tsuji, Kandai Nozu, Tadashi Sofue, Shigeo Hara, Keita Nakanishi, Tomohiko Yamamura, Shogo Minamikawa, Yoshimi Nozu, Hiroshi Kaito, Junya Fujimura, Tomoko Horinouchi, Naoya Morisada, Ichiro Morioka, Mariko Ikeda, Masafumi Matsuo, Kazumoto Iijima

研究成果: Article

1 引用 (Scopus)

抄録

Background/Aims: Glomerulopathy with fibronectin deposits (GFND; OMIM: 601894) is a very rare inherited kidney disease caused by pathogenic variants in the FN1 gene. Only 9 exonic pathogenic variants in FN1, 9 at the heparin-binding site, and 1 at the integrin-binding site have been reported. No intronic variants in FN1 have been detected. Methods: We found a pathogenic intronic variant in intron 36 (c.5888-2A>G) located at the heparin-binding site. To determine whether this mutation influences splicing processes, we conducted RT-PCR analysis and an in vitro splicing assay using minigene construction. Results: RT-PCR using RNA extracted from leukocytes of the proband failed because of the low expression of FN1 mRNA in leukocytes. We conducted in vitro functional splicing analysis using minigenes and found that c.5888-2A>G caused a 12 bp deletion at exon 37 by the activation of a novel splicing acceptor site within exon 37. We were able to detect the same abnormal transcript in mRNA extracted from the patient's urinary sediment and confirmed the pathogenicity of c.5888-2A>G by both RT-PCR using the patient sample and an in vitro splicing assay. Conclusion: Intronic variants can cause GFND. Minigene analysis is useful for determining the pathogenicity of the intronic variants and could be used for all inherited kidney diseases.

元の言語English
ページ(範囲)166-171
ページ数6
ジャーナルNephron
138
発行部数2
DOI
出版物ステータスPublished - 01-02-2018

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Binding Sites
Kidney Diseases
Polymerase Chain Reaction
Virulence
Heparin
Exons
Leukocytes
Genetic Databases
Messenger RNA
Integrins
Introns
RNA
Mutation
Genes
Glomerulopathy with fibronectin deposits
In Vitro Techniques
1,7,9,11-tetrahydroxy-3-methyl-8,13-dioxo-5,6,8,13-tetrahydrobenzo(a)tetracene-2-carboxylic acid

All Science Journal Classification (ASJC) codes

  • Physiology
  • Nephrology
  • Physiology (medical)
  • Urology

これを引用

Tsuji, Y., Nozu, K., Sofue, T., Hara, S., Nakanishi, K., Yamamura, T., ... Iijima, K. (2018). Detection of a splice site variant in a patient with glomerulopathy and fibronectin deposits. Nephron, 138(2), 166-171. https://doi.org/10.1159/000484209
Tsuji, Yurika ; Nozu, Kandai ; Sofue, Tadashi ; Hara, Shigeo ; Nakanishi, Keita ; Yamamura, Tomohiko ; Minamikawa, Shogo ; Nozu, Yoshimi ; Kaito, Hiroshi ; Fujimura, Junya ; Horinouchi, Tomoko ; Morisada, Naoya ; Morioka, Ichiro ; Ikeda, Mariko ; Matsuo, Masafumi ; Iijima, Kazumoto. / Detection of a splice site variant in a patient with glomerulopathy and fibronectin deposits. :: Nephron. 2018 ; 巻 138, 番号 2. pp. 166-171.
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Tsuji, Y, Nozu, K, Sofue, T, Hara, S, Nakanishi, K, Yamamura, T, Minamikawa, S, Nozu, Y, Kaito, H, Fujimura, J, Horinouchi, T, Morisada, N, Morioka, I, Ikeda, M, Matsuo, M & Iijima, K 2018, 'Detection of a splice site variant in a patient with glomerulopathy and fibronectin deposits', Nephron, 巻. 138, 番号 2, pp. 166-171. https://doi.org/10.1159/000484209

Detection of a splice site variant in a patient with glomerulopathy and fibronectin deposits. / Tsuji, Yurika; Nozu, Kandai; Sofue, Tadashi; Hara, Shigeo; Nakanishi, Keita; Yamamura, Tomohiko; Minamikawa, Shogo; Nozu, Yoshimi; Kaito, Hiroshi; Fujimura, Junya; Horinouchi, Tomoko; Morisada, Naoya; Morioka, Ichiro; Ikeda, Mariko; Matsuo, Masafumi; Iijima, Kazumoto.

:: Nephron, 巻 138, 番号 2, 01.02.2018, p. 166-171.

研究成果: Article

TY - JOUR

T1 - Detection of a splice site variant in a patient with glomerulopathy and fibronectin deposits

AU - Tsuji, Yurika

AU - Nozu, Kandai

AU - Sofue, Tadashi

AU - Hara, Shigeo

AU - Nakanishi, Keita

AU - Yamamura, Tomohiko

AU - Minamikawa, Shogo

AU - Nozu, Yoshimi

AU - Kaito, Hiroshi

AU - Fujimura, Junya

AU - Horinouchi, Tomoko

AU - Morisada, Naoya

AU - Morioka, Ichiro

AU - Ikeda, Mariko

AU - Matsuo, Masafumi

AU - Iijima, Kazumoto

PY - 2018/2/1

Y1 - 2018/2/1

N2 - Background/Aims: Glomerulopathy with fibronectin deposits (GFND; OMIM: 601894) is a very rare inherited kidney disease caused by pathogenic variants in the FN1 gene. Only 9 exonic pathogenic variants in FN1, 9 at the heparin-binding site, and 1 at the integrin-binding site have been reported. No intronic variants in FN1 have been detected. Methods: We found a pathogenic intronic variant in intron 36 (c.5888-2A>G) located at the heparin-binding site. To determine whether this mutation influences splicing processes, we conducted RT-PCR analysis and an in vitro splicing assay using minigene construction. Results: RT-PCR using RNA extracted from leukocytes of the proband failed because of the low expression of FN1 mRNA in leukocytes. We conducted in vitro functional splicing analysis using minigenes and found that c.5888-2A>G caused a 12 bp deletion at exon 37 by the activation of a novel splicing acceptor site within exon 37. We were able to detect the same abnormal transcript in mRNA extracted from the patient's urinary sediment and confirmed the pathogenicity of c.5888-2A>G by both RT-PCR using the patient sample and an in vitro splicing assay. Conclusion: Intronic variants can cause GFND. Minigene analysis is useful for determining the pathogenicity of the intronic variants and could be used for all inherited kidney diseases.

AB - Background/Aims: Glomerulopathy with fibronectin deposits (GFND; OMIM: 601894) is a very rare inherited kidney disease caused by pathogenic variants in the FN1 gene. Only 9 exonic pathogenic variants in FN1, 9 at the heparin-binding site, and 1 at the integrin-binding site have been reported. No intronic variants in FN1 have been detected. Methods: We found a pathogenic intronic variant in intron 36 (c.5888-2A>G) located at the heparin-binding site. To determine whether this mutation influences splicing processes, we conducted RT-PCR analysis and an in vitro splicing assay using minigene construction. Results: RT-PCR using RNA extracted from leukocytes of the proband failed because of the low expression of FN1 mRNA in leukocytes. We conducted in vitro functional splicing analysis using minigenes and found that c.5888-2A>G caused a 12 bp deletion at exon 37 by the activation of a novel splicing acceptor site within exon 37. We were able to detect the same abnormal transcript in mRNA extracted from the patient's urinary sediment and confirmed the pathogenicity of c.5888-2A>G by both RT-PCR using the patient sample and an in vitro splicing assay. Conclusion: Intronic variants can cause GFND. Minigene analysis is useful for determining the pathogenicity of the intronic variants and could be used for all inherited kidney diseases.

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Tsuji Y, Nozu K, Sofue T, Hara S, Nakanishi K, Yamamura T その他. Detection of a splice site variant in a patient with glomerulopathy and fibronectin deposits. Nephron. 2018 2 1;138(2):166-171. https://doi.org/10.1159/000484209