Contribution of aberrant splicing to the onset mechanisms in inherited kidney diseases

• Project/Area Number: 19K08726
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53040:Nephrology-related
• Research Institution: Kobe University
• Principal Investigator: Kandai Nozu 神戸大学, 医学研究科, 教授 (70362796)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: splicing / 核酸医薬 / アンチセンスオリゴ / minigene / RNA sequence / アンチセンス治療薬

Outline of Research at the Start

１．スプライシング異常をきたしうる変異の同定およびその頻度の解明を行う：保有する遺伝情報の中から、イントロン変異やサイレント変異およびその病原性が不明なミスセンス変異の中でスプライシング異常の可能性がある変異を検出し、スプライシング異常の頻度およびその重症度への影響を明らかにする。２．スプライシング異常発症機序の解明を行う： minigeneや患者腎由来培養細胞を用いた解析系を用いて、スプライシング促進因子の発現調節を行い、その影響を調べることでスプライシング異常発生機序を解明する。３．新規疾患特異的治療法の開発を行う：すべての遺伝性腎疾患を対象とした治療法の開発を行う。

Outline of Final Research Achievements

We have investigated in vitro, in vivo, and clinical viewpoints that splicing abnormalities in the pathogenesis mechanism and severity of various hereditary renal diseases are greatly involved. As a result, the substitution of the last base of the exon, which has been interpreted as a missense variant, causes a splicing abnormality, and as a result, the analysis of the genotype-phenotype correlation has been conducted based on incorrect information. It was also clarified that some hereditary renal diseases can be treated by controlling splicing. We have succeeded in finding a lot of important information that will lead to the development of therapeutic methods using nucleic acid drugs.

Academic Significance and Societal Importance of the Research Achievements

これまで遺伝性腎疾患においてスプライシング異常の関与はほとんど検討されてきていなかったが、今回の研究から遺伝子の1塩基置換であってもその結果、ミスセンス変異ではなくスプライシング異常を来すことにより病気を発症していることが多々あることが証明できた。一般的にミスセンス変異は軽症、スプライシング異常を来す変異は重症の臨床像を呈することが多いが、ミスセンス変異と解釈することにより、誤った臨床的重症度評価をする可能性がある。本研究成果はそれに対する警鐘を鳴らす物となった。また、スプライシングを正しく理解することでスプライシングを制御による治療法の開発へとつながることも本研究で示すことができた。

Research Products

• [Journal Article] The Contribution of COL4A5 Splicing Variants to the Pathogenesis of X-Linked Alport Syndrome (2022)
  • Author(s): Yamamura Tomohiko、Horinouchi Tomoko、Aoto Yuya、Lennon Rachel、Nozu Kandai
  • Journal Title: Frontiers in Medicine Volume: 9
  • DOI: 10.3389/fmed.2022.841391
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clinical features of autosomal recessive polycystic kidney disease in the Japanese population and analysis of splicing in PKHD1 gene for determination of phenotypes (2022)
  • Author(s): Ishiko S, Morisada N, Kondo A, Nagai S, Aoto Y, Okada E, Rossanti R, Sakakibara N, Nagano C, Horinouchi T, Yamamura T, Ninchoji T, Kaito H, Hamada R, Shima Y, Nakanishi K, Matsuo M, Iijima K, Nozu K.
  • Journal Title: Clin Exp Nephrol . Volume: 26 Issue: 2 Pages: 140-153
  • DOI: 10.1007/s10157-021-02135-3
  • NAID: 120007188940
  • Peer Reviewed / Open Access
• [Journal Article] Clear Evidence of <i>LAMA5</i> Gene Biallelic Truncating Variants Causing Infantile Nephrotic Syndrome (2021)
  • Author(s): Taniguchi Yukimasa、Nagano China、Sekiguchi Kiyotoshi、Tashiro Atsushi、Sugawara Noriko、Sakaguchi Haruhide、Umeda Chisato、Aoto Yuya、Ishiko Shinya、Rossanti Rini、Sakakibara Nana、Horinouchi Tomoko、Yamamura Tomohiko、Kondo Atsushi、Nagai Sadayuki、Nagase Hiroaki、Iijima Kazumoto、Miner Jeffrey H.、Nozu Kandai
  • Journal Title: Kidney360 Volume: 2 Issue: 12 Pages: 1968-1978
  • DOI: 10.34067/kid.0004952021
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Last Nucleotide Substitutions of COL4A5 Exons Cause Aberrant Splicing (2021)
  • Author(s): Aoto Y, Horinouchi T, Yamamura T, Kondo A, Nagai S, Ishiko S, Okada E, Rossanti R, Sakakibara N, Nagano C, Awano H, Nagase H, Shima Y, Nakanishi K, Matsuo M, Iijima K, Nozu K.
  • Journal Title: Kidney Int Rep. Volume: 7 Issue: 1 Pages: 108-116
  • DOI: 10.1016/j.ekir.2021.10.012
  • NAID: 120007188929
  • Peer Reviewed / Open Access
• [Journal Article] A disease-causing variant of COL4A5 in a Chinese family with Alport syndrome: a case series (2021)
  • Author(s): Wu Jing、Zhang Jun、Liu Li、Zhang Bo、Yamamura Tomohiko、Nozu Kandai、Matsuo Masafumi、Zhao Jinghong
  • Journal Title: BMC Nephrology Volume: 22 Issue: 1
  • DOI: 10.1186/s12882-021-02585-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Systematic Review of Genotype-Phenotype Correlations in Frasier Syndrome (2021)
  • Author(s): Tsuji Y, Yamamura T, Nagano C, Horinouchi T, Sakakibara N, Ishiko S, Aoto Y, Rossanti R, Okada E, Tanaka E, Tsugawa K, Okamoto T, Sawai T, Araki Y, Shima Y, Nakanishi K, Nagase H, Matsuo M, Iijima K, Nozu K.
  • Journal Title: Kidney Int Rep . Volume: 6 Issue: 10 Pages: 2585-2593
  • DOI: 10.1016/j.ekir.2021.07.010
  • NAID: 120007165718
  • Peer Reviewed / Open Access
• [Journal Article] Usefulness of functional splicing analysis to confirm precise disease pathogenesis in Diamond-Blackfan anemia caused by intronic variants in <i>RPS19</i> (2021)
  • Author(s): Takafuji Satoru、Mori Takeshi、Nishimura Noriyuki、Yamamoto Nobuyuki、Uemura Suguru、Nozu Kandai、Terui Kiminori、Toki Tsutomu、Ito Etsuro、Muramatsu Hideki、Takahashi Yoshiyuki、Matsuo Masafumi、Yamamura Tomohiko、Iijima Kazumoto
  • Journal Title: Pediatric Hematology and Oncology Volume: 38 Issue: 6 Pages: 515-527
  • DOI: 10.1080/08880018.2021.1887984
  • Peer Reviewed / Open Access
• [Journal Article] An extremely mild clinical course in a case with LAMB2-associated nephritis diagnosed with next-generation sequencing. (2021)
  • Author(s): Sakuraya K, Nozu K, Murakami H, Nagano C, Horinouchi T, Fujinaga S, Iijima K, Ohtomo Y.
  • Journal Title: CEN Case Reports Volume: - Issue: 3 Pages: 359-363
  • DOI: 10.1007/s13730-021-00574-1
  • Peer Reviewed / Open Access
• [Journal Article] Utility of glomerular Gd-IgA1 staining for indistinguishable cases of IgA nephropathy or Alport syndrome (2021)
  • Author(s): Ishiko Shinya、Tanaka Akihito、Takeda Asami、Hara Masayuki、Hamano Naoto、Koizumi Masahiro、Ueno Toshinori、Hayashi Hiroki、Kondo Atsushi、Nagai Sadayuki、Aoto Yuya、Nozu Kandai
  • Journal Title: Clinical and Experimental Nephrology Volume: Epub Issue: 7 Pages: 779-787
  • DOI: 10.1007/s10157-021-02054-3
  • NAID: 120007125745
  • Peer Reviewed / Open Access
• [Journal Article] Genetic background, recent advances in molecular biology, and development of novel therapy in Alport syndrome (2020)
  • Author(s): Nozu Kandai、Takaoka Yutaka、Kai Hirofumi、Takasato Minoru、Yabuuchi Kensuke、Yamamura Tomohiko、Horinouchi Tomoko、Sakakibara Nana、Ninchoji Takeshi、Nagano China、Iijima Kazumoto
  • Journal Title: Kidney Research and Clinical Practice Volume: 39 Issue: 4 Pages: 402-413
  • DOI: 10.23876/j.krcp.20.111
  • NAID: 120006956375
  • Peer Reviewed / Open Access
• [Journal Article] Genotype-phenotype correlations influence the response to angiotensin-targeting drugs in Japanese patients with male X-linked Alport syndrome (2020)
  • Author(s): Yamamura Tomohiko、Horinouchi Tomoko、Nagano China、Omori Takashi、Sakakibara Nana、Aoto Yuya、Ishiko Shinya、Nakanishi Koichi、Shima Yuko、Nagase Hiroaki、Takeda Hiroki、Rossanti Rini、Ye Ming Juan、Nozu Yoshimi、Ishimori Shingo、Ninchoji Takeshi、Kaito Hiroshi、Morisada Naoya、Iijima Kazumoto、Nozu Kandai
  • Journal Title: Kidney International Volume: 98 Issue: 6 Pages: 1605-1614
  • DOI: 10.1016/j.kint.2020.06.038
  • Peer Reviewed / Open Access
• [Journal Article] Comparison of clinical and genetic characteristics between Dent disease 1 and Dent disease 2 (2020)
  • Author(s): Sakakibara Nana、Nagano China、Ishiko Shinya、Horinouchi Tomoko、Yamamura Tomohiko、Minamikawa Shogo、Shima Yuko、Nakanishi Koichi、Ishimori Shingo、Morisada Naoya、Iijima Kazumoto、Nozu Kandai
  • Journal Title: Pediatric Nephrology Volume: 35 Issue: 12 Pages: 2319-2326
  • DOI: 10.1007/s00467-020-04701-5
  • NAID: 120007052636
  • Peer Reviewed
• [Journal Article] Onset mechanism of a female patient with Dent disease 2 (2020)
  • Author(s): Okamoto Takayuki、Sakakibara Nana、Nozu Kandai、Takahashi Toshiyuki、Hayashi Asako、Sato Yasuyuki、Nagano China、Matsuo Masafumi、Iijima Kazumoto、Manabe Atsushi
  • Journal Title: Clinical and Experimental Nephrology Volume: 24 Issue: 10 Pages: 946-954
  • DOI: 10.1007/s10157-020-01926-4
  • NAID: 120007160299
  • Peer Reviewed
• [Journal Article] Common risk variants in NPHS1 and TNFSF15 are associated with childhood steroid-sensitive nephrotic syndrome (2020)
  • Author(s): Jia Xiaoyuan、Yamamura Tomohiko、et al.
  • Journal Title: Kidney International Volume: 98 Issue: 5 Pages: 1308-1322
  • DOI: 10.1016/j.kint.2020.05.029
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Pathogenic evaluation of synonymous COL4A5 variants in X‐linked Alport syndrome using a minigene assay (2020)
  • Author(s): Horinouchi Tomoko、Yamamura Tomohiko、Minamikawa Shogo、Nagano China、Sakakibara Nana、Nakanishi Koichi、Shima Yuko、Morisada Naoya、Ishiko Shinya、Aoto Yuya、Nagase Hiroaki、Takeda Hiroki、Rossanti Rini、Ishimori Shingo、Kaito Hiroshi、Matsuo Masafumi、Iijima Kazumoto、Nozu Kandai
  • Journal Title: Molecular Genetics & Genomic Medicine Volume: 8 Issue: 8
  • DOI: 10.1002/mgg3.1342
  • NAID: 120006879547
  • Peer Reviewed / Open Access
• [Journal Article] Development of an Exon Skipping Therapy for X-linked Alport Syndrome With Truncating Variants in COL4A5. (2020)
  • Author(s): Yamamura T, Horinouchi T, Adachi T, Terakawa M, Takaoka Y, Omachi K, Takasato M, Takaishi K, Shoji T, Onishi Y, Kanazawa Y, Koizumi M, Tomono Y, Sugano S, Shono A, Minamikawa S, Nagano C, Sakakibara N, Ishiko S, Aoto Y, Kamura M, Harita Y, Miura K, Kanda S, Morisada N, and 6 others.
  • Journal Title: Nat Commun . Volume: 11(1) Issue: 1 Pages: 2777-2777
  • DOI: 10.1038/s41467-020-16605-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Trimerization and genotype-phenotype correlation of COL4A5 mutants in Alport syndrome (2020)
  • Author(s): Kamura Misato、Yamamura Tomohiko、Omachi Kohei、Suico Mary Ann、Nozu Kandai、Kaseda Shota、Kuwazuru Jun、Shuto Tsuyoshi、Iijima Kazumoto、Kai Hirofumi
  • Journal Title: Kidney International Reports Volume: - Issue: 5 Pages: 718-726
  • DOI: 10.1016/j.ekir.2020.01.008
  • NAID: 120006818746
  • Peer Reviewed / Open Access
• [Journal Article] Functional analysis of suspected splicing variants in CLCN5 gene in Dent disease 1. (2020)
  • Author(s): Inoue T, Nagano C, Matsuo M, Yamamura T, Sakakibara N, Horinouchi T, Shibagaki Y, Ichikawa D, Aoto Y, Ishiko S, Ishimori S, Rossanti R, Iijima K, Nozu K.
  • Journal Title: Clin Exp Nephrol Volume: in press Issue: 7 Pages: 606-612
  • DOI: 10.1007/s10157-020-01876-x
  • NAID: 120007028963
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive genetic diagnosis of Japanese patients with severe proteinuria. (2020)
  • Author(s): Nagano C, Yamamura T, Horinouchi T, Aoto Y, Ishiko S, Sakakibara N, Shima Y, Nakanishi K, Nagase H, Iijima K, Nozu K.
  • Journal Title: Sci Rep. Volume: 10(1) Issue: 1 Pages: 428-437
  • DOI: 10.1038/s41598-019-57149-5
  • NAID: 120006884200
  • Peer Reviewed / Open Access
• [Journal Article] Molecular mechanisms determining severity in patients with Pierson syndrome (2020)
  • Author(s): Minamikawa Shogo、Miwa Saori、Inagaki Tetsuji、Nishiyama Kei、Kaito Hiroshi、Ninchoji Takeshi、Yamamura Tomohiko、Nagano China、Sakakibara Nana、Ishimori Shingo、Hara Shigeo、Yoshikawa Norishige、Hirano Daishi、Harada Ryoko、Hamada Riku、Matsunoshita Natsuki、Nagata Michio、Shima Yuko、Nozu Kandai
  • Journal Title: Journal of Human Genetics Volume: 65 Issue: 4 Pages: 355-362
  • DOI: 10.1038/s10038-019-0715-0
  • NAID: 120006811107
  • Peer Reviewed
• [Journal Article] Determination of the pathogenicity of known COL4A5 intronic variants by in vitro splicing assay. (2019)
  • Author(s): Horinouchi T, Nozu K, Yamamura T, Minamikawa S, Nagano C, Sakakibara N, Nakanishi K, Shima Y, Morisada N, Ishiko S, Aoto Y, Nagase H, Takeda H, Rossanti R, Kaito H, Matsuo M, Iijima K.
  • Journal Title: Sci Rep. Volume: 9(1) Issue: 1 Pages: 12696-12696
  • DOI: 10.1038/s41598-019-48990-9
  • NAID: 120006825910
  • Peer Reviewed / Open Access
• [Journal Article] Comparison between conventional and comprehensive sequencing approaches for genetic diagnosis of Alport syndrome. (2019)
  • Author(s): Yamamura T, Nozu K, Minamikawa S, Horinouchi T, Sakakibara N, Nagano C, Aoto Y, Ishiko S, Nakanishi K, Shima Y, Nagase H, Rossanti R, Ye MJ, Nozu Y, Ishimori S, Morisada N, Kaito H, Iijima K.
  • Journal Title: Mol Genet Genomic Med. Volume: 7(9) Issue: 9
  • DOI: 10.1002/mgg3.883
  • NAID: 120006733124
  • Peer Reviewed / Open Access
• [Journal Article] Inherited salt‐losing tubulopathy: an old condition but a new category of tubulopathy (2019)
  • Author(s): Nozu Kandai、Yamamura Tomohiko、Horinouchi Tomoko、Nagano China、Sakakibara Nana、Ishikura Kenji、Hamada Riku、Morisada Naoya、Iijima Kazumoto
  • Journal Title: Pediatrics International Volume: - Issue: 4 Pages: 428-437
  • DOI: 10.1111/ped.14089
  • NAID: 120006844963
  • Peer Reviewed / Open Access
• [Journal Article] Clinical and Genetic Characteristics in Patients With Gitelman Syndrome. (2018)
  • Author(s): Fujimura J, Nozu K, Yamamura T, Minamikawa S, Nakanishi K, Horinouchi T, Nagano C, Sakakibara N, Nakanishi K, Shima Y, Miyako K, Nozu Y, Morisada N, Nagase H, Ninchoji T, Kaito H, Iijima K.
  • Journal Title: Kidney Int Rep. Volume: ４ Issue: 1 Pages: 119-125
  • DOI: 10.1016/j.ekir.2018.09.015
  • NAID: 120006847258
  • Peer Reviewed / Open Access
• [Presentation] 遺伝性腎疾患研究最前線：国内で行われている最先端の研究に触れる (2021)
  • Author(s): 野津寛大
  • Organizer: 第64回日本腎臓学会学術総会
  • Invited
• [Presentation] Participation in "KDIGO in Genetics" working group as a researcher (2021)
  • Author(s): 野津寛大
  • Organizer: 第64回日本腎臓学会学術総会
  • Invited
• [Presentation] 4型コラーゲン関連腎症の発症機序と核酸医薬を用いたエクソンスキッピング療法の開発 (2021)
  • Author(s): 野津寛大
  • Organizer: 第53回日本結合組織学会
  • Invited
• [Presentation] Bartter syndrome and Gitelman syndrome (2021)
  • Author(s): Kandai Nozu
  • Organizer: 14th Asian Congress of Pediatric Nephrology
  • Int'l Joint Research / Invited
• [Presentation] 14.Novel Exon Skipping therapy in Alport’s Syndrome to prevent CKD and ESRD (2021)
  • Author(s): Kandai Nozu
  • Organizer: 9th Annual Emerging Trands in Dialysis Care
  • Int'l Joint Research / Invited
• [Presentation] Bartter syndrome and Gitelman syndrome (2021)
  • Author(s): Nozu K
  • Organizer: 14th Asian Congress of Pediatric Nephrology
  • Int'l Joint Research / Invited
• [Presentation] Exon skipping therapy in Alport syndrome (2020)
  • Author(s): Nozu K
  • Organizer: The American Society of Nephrology Annual Meeting
  • Int'l Joint Research / Invited
• [Presentation] Alport syndrome Up To Date and development of gene targeted therapy (2020)
  • Author(s): Nozu K
  • Organizer: Korean Society of Nephrology, virtual meeting
  • Int'l Joint Research / Invited
• [Presentation] これからの小児科学研究をどのように進めるか；活躍する先輩小児科physician-scientistからのメッセ－ジ (2019)
  • Author(s): 野津寛大
  • Organizer: 第122回日本小児科学会学術集会
  • Invited
• [Presentation] 遺伝性腎疾患に対するプレシジョンメディスン アルポート症候群に対する遺伝子標的療法の開発 (2019)
  • Author(s): 野津寛大
  • Organizer: 第54回日本小児腎臓病学会学術集会
  • Invited
• [Presentation] ゲノムから見た腎臓病 症例から考える遺伝性尿細管疾患 (2019)
  • Author(s): 野津寛大
  • Organizer: 第62回日本腎臓学会学術集会
  • Invited
• [Presentation] Bartter syndrome (2019)
  • Author(s): Kandai Nozu
  • Organizer: International Pediartic Nephrology Association 2019
  • Int'l Joint Research / Invited
• [Presentation] 9.Genetics and development of exon-skipping therapy for Alport Syndrome. (2019)
  • Author(s): Kandai Nozu
  • Organizer: International Pediartic Nephrology Association 2019
  • Int'l Joint Research / Invited