Establishment of new classification for structural variation based on next generation sequencing

2015 Fiscal Year Annual Research Report

• Project/Area Number: 15H04710
• Research Institution: Fujita Health University
• Principal Investigator: 倉橋 浩樹 藤田保健衛生大学, 総合医科学研究所, 教授 (30243215)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: ゲノム / 染色体 / 端部欠失 / 逆位重複 / 複製の停止 / 鋳型乗り換え / FoSTeS / MMBIR

Outline of Annual Research Achievements

染色体端部欠失・逆位重複の発生メカニズムはまだよくわかっていない。頻度の高い8pの端部欠失・逆位重複は相同性の高いinverted repeat配列を介した非アリル間相同組み換え（Non-Allelic Homologous Recombination: NAHR）とされているが、そうでない散発性のものに関しては未解明である。５例の染色体端部欠失・逆位重複の切断点の解析を行った。4p, 8q, 9p, 10q, 11qの染色体端部欠失の症例にマイクロアレイ染色体検査を行うことで端部欠失のすぐ近くに接して重複領域が存在することが同定され、FISH法を用いた向きの解析により逆位重複が確定された。引き続いて、次世代シーケンサーによる全ゲノムシーケンスをメイトペアで行うことにより、端部欠失・逆位重複の切断点及びジャンクションの配列情報を得た。５例のすべての症例において逆位の折り返し点には対称性はなく、長い介在配列が存在した。介在配列のサイズは共通して数kbであった。ジャンクション配列は数塩基のマイクロホモロジーを介して結合しており、なんらかの複製の停止（Fork Stalling and Template Switching: FoSTeS）の関与が推測された。停止後の複製の再開は、DNA端の核内距離が近い同時進行中の近傍の複製フォークへの侵入、鋳型乗り換え（Microhomology-Mediated Break-Induced Replication: MMBIR）による複製の再開が示唆されているが、本研究の結果からは、同一フォーク内のleading鎖からlagging鎖への鋳型乗り換えが生じたときに、逆位重複が生じることが推測された。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

染色体端部欠失・逆位重複の切断点の解析を行い、その発生メカニズムの一端が解明できた。

Strategy for Future Research Activity

次の研究対象である染色体挿入の発生メカニズムの解明に全力を傾ける。方法は、同じく、次世代シーケンサーによる全ゲノムシーケンスをメイトペアで行うことにより、切断点及びジャンクションの配列情報を得ることで、発生メカニズムに迫る。

Research Products

• [Journal Article] PCS/MVA syndrome caused by an Alu insertion in the BUB1B gene. (2017)
  • Author(s): Kato M, Kato T, Hosoba E, Ohashi M, Fujisaki M, Ozaki M, Yamaguchi M, Sameshima H, *Kurahashi H.
  • Journal Title: Hum Genome Var Volume: 印刷中 Pages: 印刷中
  • DOI: nd
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A Family With Craniofrontonasal Syndrome: The First Report of Familial Cases of Craniofrontonasal Syndrome With Bilateral Cleft Lip and Palate. (2017)
  • Author(s): Inoue Y, Sakamoto Y, Sugimoto M, Inagaki H, Boda H, Miyata M, Kato H, Kurahashi H, *Okumoto T.
  • Journal Title: Cleft Palate Craniofac J Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1597/15-347
• [Journal Article] Palindrome-mediated translocations in humans: a new mechanistic model for gross chromosomal rearrangements. (2016)
  • Author(s): Inagaki H, Kato T, Tsutsumi M, Ouchi Y, Ohye T, *Kurahashi H.
  • Journal Title: Front Genet Volume: 7 Pages: 125
  • DOI: 10.3389/fgene.2016.00125
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Novel compound heterozygous variants in PLK4 identified in a patient with autosomal recessive microcephaly and chorioretinopathy. (2016)
  • Author(s): Tsutsumi M, Yokoi S, Miya F, Miyata M, Kato M, Okamoto N, Tsunoda T, Yamasaki M, Kanemura Y, Kosaki K, Saitoh S, *Kurahashi H.
  • Journal Title: Eur J Hum Genet Volume: 24 Pages: 1702-1706
  • DOI: 10.1038/ejhg.2016.119
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A PDE3A mutation in familial hypertension and brachydactyly syndrome. (2016)
  • Author(s): Boda H, Uchida H, Takaiso N, Ouchi Y, Fujita N, Kuno A, Hata T, Nagatani A, Funamoto Y, Miyata M, Yoshikawa T, *Kurahashi H, Inagaki H.
  • Journal Title: J Hum Genet Volume: 61 Pages: 701-3
  • DOI: 10.1038/jhg.2016.32
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Next-generation sequencing discloses a nonsense mutation in the dystrophin gene from long preserved dried umbilical cord and low-level somatic mosaicism in the proband mother. (2016)
  • Author(s): *Taniguchi-Ikeda M, Takeshima Y, Lee T, Nishiyama M, Awano H, Yagi M, Unzaki A, Nozu K, Nishio H, Matsuo M, Kurahashi H, Toda T, Morioka I, Iijima K.
  • Journal Title: J Hum Genet Volume: 61 Pages: 351-5
  • DOI: 10.1038/jhg.2015.157
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Annexin A5 haplotype M2 is not a risk factor for recurrent miscarriages in Northern Europe, is there sufficient evidence? (2016)
  • Author(s): *Markoff A, Kurahashi H, Grandone E, Bogdanova N.
  • Journal Title: Reprod Biomed Online Volume: 32 Pages: 469-73
  • DOI: 10.1016/j.rbmo.2016.02.004
  • Peer Reviewed
• [Journal Article] Authors' response to the letter of Nagirnaja et al., "Response to annexin A5 haplotype M2 is not a risk factor for recurrent miscarriages in Northern Europe, is there sufficient evidence?" (2016)
  • Author(s): Markoff A, Kurahashi H, Grandone E, Bogdanova N.
  • Journal Title: Reprod Biomed Online Volume: 33 Pages: 116-7
  • DOI: 10.1016/j.rbmo.2016.05.006
  • Peer Reviewed
• [Journal Article] A simple cytogenetic method to detect chromosomally integrated human herpesvirus-6. (2016)
  • Author(s): Ohye T, Kawamura Y, Inagaki H, Yoshikawa A, Ihira M, Yoshikawa T, *Kurahashi H.
  • Journal Title: J Virol Methods Volume: 228 Pages: 74-8
  • DOI: 10.1016/j.jviromet.2015.11.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Complex X-Chromosomal Rearrangements in Two Women with Ovarian Dysfunction: Implications of Chromothripsis/Chromoanasynthesis-Dependent and -Independent Origins of Complex Genomic Alterations. (2016)
  • Author(s): Suzuki E, Shima H, Toki M, Hanew K, Matsubara K, Kurahashi H, Narumi S, Ogata T, Kamimaki T, *Fukami M.
  • Journal Title: Cytogenet Genome Res Volume: 150 Pages: 86-92
  • DOI: 10.1159/000455026
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Increased levels of soluble corin in pre-eclampsia and fetal growth restriction. (2016)
  • Author(s): Miyazaki J, *Nishizawa H, Kambayashi A, Ito M, Noda Y, Terasawa S, Kato T, Miyamura H, Shiogama K, Sekiya T, Kurahashi H, Fujii T.
  • Journal Title: Placenta Volume: 48 Pages: 20-25
  • DOI: 10.1016/j.placenta.2016.10.002
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Successful living donor liver transplantation for classical maple syrup urine disease. (2016)
  • Author(s): Yasui T, Suzuki T, Hara F, Watanabe S, Uga N, Naoe A, Yoshikawa T, Ito T, Nakajima Y, Miura H, Sugioka A, Kato Y, Tokoro T, Tanahashi Y, Kasahara M, Fukuda A, Kurahashi H. Successful living donor liver transplantation for classical maple syrup urine disease.
  • Journal Title: Pediatr Transplant Volume: 20 Pages: 707-710
  • DOI: 10.1111/petr.12738
  • Peer Reviewed
• [Journal Article] Mutation analysis of the JUNO gene in female infertility of unknown etiology. (2016)
  • Author(s): Takaiso N, Nishizawa H, Nishiyama S, Sawada T, Hosoba E, Ohye T, Sato T, Inagaki H, *Kurahashi H.
  • Journal Title: Fujita Med J Volume: 2 Pages: 59-61
  • Peer Reviewed
• [Presentation] Gene expression analysis reveals molecular phenotypes related to schizophrenia in Neurogranin knockout mice. (2016)
  • Author(s): Hattori S, Hagihara H, Kameyama T, Ouchi Y, Inagaki H, Kurahashi H, Huang FL, Huang KP, Miyakawa T.
  • Organizer: Neuroscience 2016
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-11-12 – 2016-11-16
  • Int'l Joint Research
• [Presentation] Missense mutations in the PLK4 gene identified in a patient with autosomal recessive microcephaly and chorioretinopathy. (2016)
  • Author(s): Tsutsumi M, Yokoi S, Miya F, Miyata M, Kato M, Okamoto N, Tsunoda T, Yamasaki M, Kanemura Y, Kosaki K, Saitoh S, Kurahashi H.
  • Organizer: ASHG2016
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2016-10-18 – 2016-10-22
  • Int'l Joint Research
• [Presentation] Mechanistic analysis and prediction of interchromosomal insertional translocation. (2016)
  • Author(s): Kato T, Ouchi Y, Inagaki H, Makita Y, Mizuno S, Kurahashi H.
  • Organizer: ASHG2016
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2016-10-18 – 2016-10-22
  • Int'l Joint Research
• [Presentation] 13q22.2q34 tetrasomy mosaicism due to an inverted duplication with a neocentromere. (2016)
  • Author(s): Hattori T, Togawa T, Togawa Y, Kawabe H, Kato T, Kurahashi H, Saitoh S, Kouwaki M, Koyama N.
  • Organizer: ASHG2016
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2016-10-18 – 2016-10-22
  • Int'l Joint Research
• [Presentation] Relationship between various clinical features in a genotyped population investigated for tuberous sclerosis complex. (2016)
  • Author(s): Ishihara N, Sasaki H, Kato T, Inagaki H, Tsutsumi M, Shiroki R, Kurahashi H.
  • Organizer: ASHG2016
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2016-10-18 – 2016-10-22
  • Int'l Joint Research
• [Presentation] Mechanism for Structural Variation. (2016)
  • Author(s): Kurahashi H.
  • Organizer: APAC RGH Symposium Singapore
  • Place of Presentation: Singapore
  • Year and Date: 2016-10-04 – 2016-10-04
  • Int'l Joint Research / Invited
• [Presentation] De novo translocation frequency of the recurrent constitutional t(11;22)(q23;q11.2) in normal somatic tissues. (2016)
  • Author(s): Kurahashi H, Kato T, Suzuki M, Tsutsumi M, Ohye T, Inagaki H.
  • Organizer: ESHG2016
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2016-05-21 – 2016-05-24
  • Int'l Joint Research
• [Presentation] Palindrome-mediated recurrent translocations in humans. (2016)
  • Author(s): Kurahashi H.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research / Invited
• [Presentation] Mechanisms of interchromosomal insertional translocation. (2016)
  • Author(s): Kato T, Ouchi Y, Inagaki H, Makita Y, Mizuno S, Kurahashi H.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] A PDE3A mutation in familial hypertension and brachydactyly syndrome. (2016)
  • Author(s): Inagaki H, Boda H, Uchida H, Takaiso N, Ouchi Y, Fujita N, Kuno A, Hata T, Nagatani A, Funamoto Y, Miyata M, Yoshikawa T, Kurahashi H.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Spermatogenic failure by impaired meiotic sex chromosome inactivation in a mouse with reciprocal translocation. (2016)
  • Author(s): Tsutsumi M, Kato T, Inagaki H, Ohye T, Kurahashi H.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Cell culture model for X-linked disorder: craniofrontonasal dysplasia and severe phenotype in female. (2016)
  • Author(s): Sugimoto M, Inagaki H, Tsutsumi M, Inoue Y, Taguchi Y, Boda H, Miyata M, Okumoto T, Yoshikawa T, Kurahashi H.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] A child presenting distinct phenotype in severe alternating hemiplegia with a novel ATP1A3 mutation. (2016)
  • Author(s): Ishihara N, Inagaki H, Miyake M, Ouchi Y, Ohye T, Tsutsumi M, Yoshikawa T, Kurahashi H.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Sex determination of the fetus by noninvasive prenatal testing (NIPT) with maternal blood. (2016)
  • Author(s): Noda Y, Nishizawa H, Kato T, Kambayashi A, Terasawa S, Miyazaki J, Ito M, Kurahashi H, Fujii T.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Increased levels of soluble corin in patients with pre-eclampsia and fetal growth restriction. (2016)
  • Author(s): Miyazaki J, Nishizawa H, Kambayashi A, Ito M, Noda Y, Terasawa S, Kato T, Miyamura H, Sekiya T, Kurahashi H, Fujii T.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Advantage of next generation sequencing in molecular diagnosis in DMD -mutation screening with long preserved dried umbilical cord and detection of mosaicism-. (2016)
  • Author(s): Unzaki A, Taniguchi-Ikeda M, Takeshima Y, Lee T, Awano H, Yagi M, Kurahashi H, Morioka I, Toda T, Matsuo M, Iijima K.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Prenatal diagnosis of the Premature chromosome separation/ mosaic variegated aneuploidy (PCS/MVA) syndrome in fetus with microcephalus. (2016)
  • Author(s): Ohashi M, Yamaguchi M, Ishii M, Yamaguchi T, Akeno K, Fijisaki M, Sumiyoshi C, Sameshima H, Ozaki M, Kato T, Inagaki H, Kurahashi H.
  • Organizer: ICHG2016
  • Place of Presentation: Kyoto, Japan、京都国際会館
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Remarks] ようこそ分子遺伝学研究室へ
  • URL: http://molgen.icms.fujita-hu.ac.jp