Development of new treatments for bone dysplasia caused by a mutation in TGFB1 utilizing patient-derived iPS cells

• Project/Area Number: 15K10481
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Nagasaki University
• Principal Investigator: KINOSHITA Akira 長崎大学, 原爆後障害医療研究所, 講師 (60372778)
• Co-Investigator(Renkei-kenkyūsha): KINOSHITA Naoe 長崎大学, 病院(医学系), 助教 (50380928) ; YOSHIURA Koh-Ichiro 長崎大学, 原爆後障害医療研究所, 教授 (00304931)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: カムラチ-エンゲルマン病 / TGF-β1 / iPS細胞 / ゲノム編集 / カムラチ・エンゲルマン病 / 患者iPS細胞 / 骨系統疾患 / カムラチ・エンゲルマン病( / CRISPR/Cas9法

Outline of Final Research Achievements

Camurati-Engelmann disease (CAEND) is a rare, autosomal dominant, progressive diaphyseal dysplasia, which is characterized by hyperostosis and sclerosis of the diaphyses of long bones. CAEND is caused by a mutation in the gene TGFB1, encoding TGF-β1. We established induced pluripotent stem (iPS) cells derived from an individual with CAEND. These cells were easier to differentiate than those derived from a healthy individual. Since genetic background strongly affects the CAEND phenotype, we attempted to correct the arginine-to-histidine mutation at residue 218 (R218H) in TGFB1 utilizing a genome editing system. Although we successfully obtained homologous recombinant clones of HEK293T cells, we failed to obtain recombinant clones of iPS cells so far. Furthermore, we discovered new Japanese CAEND cases that had an arginine-to-cysteine mutation at residue 218 in TGFB1. We also identified a de novo mutation in a gene involved in the TGF signaling pathway, which causes hyperostosis.

Research Products

• [Journal Article] Whole-exome sequencing and gene-based rare variant association tests suggest that PLA2G4E might be a risk gene for panic disorder. Transl Psychiatry. (2018)
  • Author(s): Morimoto Y, Shimada-Sugimoto M, Otowa T, Yoshida S, Kinoshita A, Mishima H, Yamaguchi N, Mori T, Imamura A, Ozawa H, Kurotaki N, Ziegler C, Domschke K, Deckert J, Umekage T, Tochigi M, Kaiya H, Okazaki Y, Tokunaga K, Sasaki T, Yoshiura KI, Ono S.
  • Journal Title: Transl Psychiatry Volume: 8 Issue: 1 Pages: 41-41
  • DOI: 10.1038/s41398-017-0088-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clonal dynamics in a case of acute monoblastic leukemia that later developed myeloproliferative neoplasm. (2018)
  • Author(s): Sato S, Itonaga H, Taguchi M, Sawayama Y, Imanishi D, Tsushima H, Hata T, Moriuchi Y, Mishima H, Kinoshita A, Yoshiura KI, Miyazaki Y.
  • Journal Title: Int J Hematol. Volume: - Issue: 2 Pages: 213-217
  • DOI: 10.1007/s12185-018-2419-1
  • Peer Reviewed
• [Journal Article] Detection of de novo single nucleotide variants in offspring of atomic-bomb survivors close to the hypocenter by whole-genome sequencing. (2018)
  • Author(s): Horai M, Mishima H, Hayashida C, Kinoshita A, Nakane Y, Matsuo T, Tsuruda K, Yanagihara K, Sato S, Imanishi D, Imaizumi Y, Hata T, Miyazaki Y, Yoshiura KI.
  • Journal Title: J Hum Genet. Volume: 63 Issue: 3 Pages: 357-363
  • DOI: 10.1038/s10038-017-0392-9
  • Peer Reviewed
• [Journal Article] Deep sequencing reveals variations in somatic cell mosaic mutations between monozygotic twins with discordant psychiatric disease. (2017)
  • Author(s): Morimoto Y, Ono S, Imamura A, Okazaki Y, Kinoshita A, Mishima H, Nakane H, Ozawa H, Yoshiura KI, Kurotaki N.
  • Journal Title: Hum Genome Var. Volume: 4 Issue: 1 Pages: 17032-17032
  • DOI: 10.1038/hgv.2017.32
  • Peer Reviewed / Open Access
• [Journal Article] Generation and characterization of a conditional allele of Interferon Regulatory Factor 6. (2017)
  • Author(s): Smith AL, Kousa YA, Kinoshita A, Fodor K, Yang B, Schutte BC.
  • Journal Title: Genesis. Volume: 55 Issue: 7
  • DOI: 10.1002/dvg.23038
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Rapid growth of mitotically active cellular fibroma of the ovary: a case report and review of the literature. (2016)
  • Author(s): Matsuda K, Tateishi S, Akazawa Y, Kinoshita A, Yoshida S, Morisaki S, Fukushima A, Matsuwaki T, Yoshiura KI, Nakashima M.
  • Journal Title: Diagn Pathol. Volume: 11(1) Issue: 1 Pages: 101-101
  • DOI: 10.1186/s13000-016-0554-7
  • NAID: 120006987629
  • Peer Reviewed / Open Access
• [Journal Article] Expression of Somatostatin Receptor Type 2A and PTEN in Neuroendocrine Neoplasms Is Associated with Tumor Grade but Not with Site of Origin. (2016)
  • Author(s): Wada H, Matsuda K, Akazawa Y, Yamaguchi Y, Miura S, Ueki N, Kinoshita A, Yoshiura K, Kondo H, Ito M, Nagayasu T, Nakashima M.
  • Journal Title: Endocr Pathol. Volume: 27(3) Issue: 3 Pages: 179-187
  • DOI: 10.1007/s12022-016-9436-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ultra-sensitive droplet digital PCR for detecting a low-prevalence somatic GNAQ mutation in Sturge-Weber syndrome. (2016)
  • Author(s): Uchiyama Y, Nakashima M, Watanabe S, Miyajima M, Taguri M, Miyatake S, Miyake N, Saitsu H, Mishima H, Kinoshita A, Arai H, Yoshiura K, Matsumoto N.
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 22985-22985
  • DOI: 10.1038/srep22985
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Germline mutations causing familial lung cancer. (2015)
  • Author(s): Tomoshige K, Matsumoto K, Tsuchiya T, Oikawa M, Miyazaki T, Yamasaki N, Mishima H, Kinoshita A, Kubo T, Fukushima K, Yoshiura K, Nagayasu T.
  • Journal Title: Journal of Human Genetics Volume: 60 Issue: 10 Pages: 597-603
  • DOI: 10.1038/jhg.2015.75
  • Peer Reviewed
• [Journal Article] カムラチ・エンゲルマン病 (2015)
  • Author(s): 木下 晃
  • Journal Title: 日本臨床 Volume: 73 Pages: 2149-2159
• [Presentation] Mutations in the patients with NNS-like autoinflammatory diseases (2016)
  • Author(s): Akira Kinoshita
  • Organizer: International Congress of Human Genetics
  • Place of Presentation: 国立京都国際会（京都府・京都市）
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] Mutations in the patients with NNS-like autoinflammatory diseases (2016)
  • Author(s): Akira Kinoshita
  • Organizer: The 13th International Congress of Human Genetics
  • Place of Presentation: Kyoto International Conference Center (京都府・京都市）
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] 中條-西村症候群様自己炎症性疾患の変異解析 (2015)
  • Author(s): 木下 晃
  • Organizer: 第６０回日本人類遺伝学会
  • Place of Presentation: 京王プラザホテル（東京都・新宿区）
  • Year and Date: 2015-10-14