Molecular genetic analysis of congenital thrombocytopenia, hemostatic and coagulation disorder

• Project/Area Number: 19K17865
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: Yokohama City University
• Principal Investigator: UCHIYAMA Yuri 横浜市立大学, 附属病院, 助教 (50829794)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2019: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 先天性造血器障害 / 汎血球減少症 / 血小板減少症 / 全エクソーム解析 / 先天性造血障害 / 造血機能不全

Outline of Research at the Start

小児期、あるいは若年発症の造血異常や止血凝固異常症に対し、全エクソーム解析を用いた網羅的な遺伝子解析を行うことで、効率的に疾患の遺伝学的な解明を目指す。本手法を用いて既に検出した候補遺伝子に対し、その機能を明らかにし、疾患の遺伝学的な原因を明らかにすることを目指す。これらを明らかにすることで、造血不全や機能異常における、診断率の向上や治療方針決定への大きな寄与へとつながることが予測される。

Outline of Final Research Achievements

Thrombocytopenia occurring in childhood must be differentiated from immune thrombocytopenia, hematological malignancy, bone marrow failure, and inherited thrombocytopenia.
In this study, exome sequencing identified gene A and B in two families with pancytopenia and congenital thrombocytopenia, respectively. The pathogenicity of these two genes and variants were subsequently evaluated by deep-sequencing, RT-PCR, and/or RNA-sequencing. Deep-sequencing results was strongly convinced that this variant was derived from the early-phase somatic mosaicism. However, lymphoblastoid cell lines derived from patient’s blood could not include the cells with the variant. The strong toxicity of the variant of gene A induced cell mortality. A total of 42 families of congenital abnormalities of thrombosis and/or hemostasis was performed for exome sequencing. The diagnostic rate was 23.8% (10/42). Exome sequencing is useful technique for the rare disorder associated with congenital thrombocytopenia.

Academic Significance and Societal Importance of the Research Achievements

先天性血小板減少症の遺伝的原因を明らかにすることで、疾患の背景を明らかにすることができる。造血の過程のどの段階での異常かを同定することは、先天性造血障害における治療方針を検討する上で大きな指針の一つとなるため、新規疾患の遺伝的原因の解明は非常に重要である。これら先天性疾患の遺伝的疾患原因解明は、後天性疾患である多くの造血不全・造血器腫瘍の理解・治療につながることが期待される。
患者抽出方法を含め、本手法を用いた先天性血小板減少症・止血凝固異常症の遺伝的疾患病態の解明は、非常に効率的であることが示された。

Research Products

• [Journal Article] Pathogenic 12-kb copy-neutral inversion in syndromic intellectual disability identified by high-fidelity long-read sequencing (2021)
  • Author(s): Mizuguchi T, Okamoto N, Yanagihara K, Miyatake S, Uchiyama Y, Tsuchida N, Hamanaka K, Fujita A, Miyake N, Matsumoto N.
  • Journal Title: Genomics Volume: 113 Issue: 1 Pages: 1044-1053
  • DOI: 10.1016/j.ygeno.2020.10.038
  • Peer Reviewed
• [Journal Article] De novo variants in CELF2 that disrupt the nuclear localization signal cause developmental and epileptic encephalopathy (2021)
  • Author(s): Itai T, Hamanaka K, Sasaki K, Saitsu H, Miyatake S, Matsumoto N et al.
  • Journal Title: Hum Mutat Volume: 42 Issue: 1 Pages: 66-76
  • DOI: 10.1002/humu.24130
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hemizygous FLNA variant in West syndrome without periventricular nodular heterotopia (2020)
  • Author(s): Hiromoto Y, Azuma Y, Suzuki Y, Hoshina M, Uchiyama Y, Mitsuhashi S, Miyatake S, Mizuguchi T, Takata A, Miyake N, Kato M, Matsumoto N.
  • Journal Title: Hum Genome Var Volume: 7 Issue: 1 Pages: 43-43
  • DOI: 10.1038/s41439-020-00131-9
  • Peer Reviewed / Open Access
• [Journal Article] Whole exome sequencing of fetal structural anomalies detected by ultrasonography (2020)
  • Author(s): Aoi Hiromi、Mizuguchi Takeshi、Suzuki Toshifumi、Makino Shintaro、Yamamoto Yuka...Takata Atsushi、Miyake Noriko、Takeda Satoru、Itakura Atsuo、Matsumoto Naomichi
  • Journal Title: Journal of Human Genetics Volume: 66 Issue: 5 Pages: 499-507
  • DOI: 10.1038/s10038-020-00869-8
  • Peer Reviewed
• [Journal Article] Efficient detection of copy‐number variations using exome data: Batch‐ and sex‐based analyses (2020)
  • Author(s): Uchiyama Yuri、Yamaguchi Daisuke、Iwama Kazuhiro, et al.
  • Journal Title: Human Mutation Volume: 42 Issue: 1 Pages: 50-65
  • DOI: 10.1002/humu.24129
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prenatal clinical manifestations in individuals with COL4A1/2 variants (2020)
  • Author(s): Itai T, Miyatake S, Taguri M, Nakashima M, Saitsu H, Matsumoto N et al.
  • Journal Title: J Med Genet Volume: 0 Issue: 8 Pages: 106896-106896
  • DOI: 10.1136/jmedgenet-2020-106896
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Retraction Note to: Nonsense variants in STAG2 result in distinct sex-dependent phenotypes (2020)
  • Author(s): Aoi Hiromi、Lei Ming、Mizuguchi Takeshi、Nishioka Nobuko、Goto Tomohide、Miyama Sahoko、Suzuki Toshifumi、Iwama Kazuhiro、Uchiyama Yuri、Mitsuhashi Satomi、Itakura Atsuo、Takeda Satoru、Matsumoto Naomichi
  • Journal Title: Journal of Human Genetics Volume: 65 Issue: 9 Pages: 811-811
  • DOI: 10.1038/s10038-020-0782-2
  • Peer Reviewed
• [Presentation] 先天性第V因子欠乏症の遺伝学的診断とコピー数解析 (2020)
  • Author(s): 内山由理
  • Organizer: 第42回日本血栓止血学会学術集会
• [Presentation] Efficient detection of copy-number variations using whole-exome data: batch- and sex-based analyses (2020)
  • Author(s): Yuri Uchiyama, Kazuhiro Iwama, Satoko Miyatake, Kohei Hamanaka, Naomi Tsuchida, Hiromi Aoi, Yoshiteru Azuma, Toshiyuki Itai, Ken Saida, Hiromi Fukuda, Futoshi Sekiguchi, Tomohiro Sakaguchi, Sachiko Ohori, Ming Lei, Eriko Koshimizu, Atsushi Fujita, Atsushi Takata, Noriko Miyake, Takeshi Mizuguchi, Naomichi Matsumoto
  • Organizer: 日本人類遺伝学会第65回大会
• [Presentation] Molecular genetic analysis of 10 families with chronic thrombocytopenia (2019)
  • Author(s): 内山由理
  • Organizer: International Society on Thrombosis and Haemostasis
  • Int'l Joint Research
• [Presentation] von Willebrand 病の臨床診断における遺伝子解析の有用性 (2019)
  • Author(s): 内山由理
  • Organizer: 第41回日本血栓止血学会学術集会