( 報告様式 4) 16gk h0001 平成 29 年 5 月 19 日 平成 28 年度委託研究開発成果報告書 I. 基本情報 事 業 名 : ( 日本語 ) 成育疾患克服等総合研究事業 ( 英語 )Project for Baby and Infant in Research of
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1 ( 報告様式 4) 16gk h0001 平成 29 年 5 月 19 日 平成 28 年度委託研究開発成果報告書 I. 基本情報 事 業 名 : ( 日本語 ) 成育疾患克服等総合研究事業 ( 英語 )Project for Baby and Infant in Research of health and Development to Adolescent and Young adult 研究開発課題名 : ( 日本語 ) 環境 遺伝相互作用に基づく妊娠高血圧症候群の病態解明 早期診断法の開発 ( 英語 )Developmental study for early prediction and elucidation of undetermined pathological mechanism of hypertensive disorders of pregnancy based on environmental and genetic interactions. 研究開発担当者所属役職氏名 : ( 日本語 ) 東北大学東北メディカル メガバンク機構地域医療支援部門教授菅原準一 Professor Junichi Sugawara 実施期間 : 平成 28 年 4 月 1 日 ~ 平成 29 年 3 月 31 日 分担研究開発課題名 : ( 日本語 )1 三世代コホート疫学解析 2 妊婦 トリオにおける全ゲノム解析 3プロテオーム解析によるバイオマーカー探索 検証 4メタボローム解析によるバイオマーカー探索 検証 5オミックス統合情報解析 ( 英語 )1Epidemiologic analysis for Birth and Three generation cohort study. 2Whole genome analysis for familial trios including pregnant women. 3Discovery and validation of biomarkers for early prediction by proteomic analysis. 4Discovery and validation of biomarkers for early prediction by metabolomic analysis. 5Integrated bioinformatic analysis for multi-omics study. 1
2 研究開発分担者 所属役職氏名 : ( 日本語 ) 東北大学東北メディカル メガバンク機構予防医学 疫学部門教授栗山進一 Professor Shinichi Kuriyama ( 日本語 ) 東北大学東北メディカル メガバンク機構ゲノム解析部門教授安田純 Professor Jun Yasuda 研究開発分担者 ( 日本語 ) 東北大学大学院薬学研究科教授寺崎哲也所属役職氏名 : ( 英語 )Graduate school of Pharmaceutical Sciences, Tohoku University Professor Tetsuya Terasaki ( 日本語 ) 東北大学東北メディカル メガバンク機構ゲノム解析部門准教授小柴生造 Associate professor Seizo Koshiba ( 日本語 ) 東北大学東北メディカル メガバンク機構ゲノム解析部門教授長﨑正朗 Professor Masao Nagasaki 2
3 II. 成果の概要 ( 総括研究報告 ) 東北メディカル メガバンク機構 (Tohoku Medical Megabank Organization: ToMMo) は 世界的にユニークな出生三世代コホート調査を推進し 統合バイオバンクを構築している 本研究は 妊娠高血圧症候群の新しい早期診断方法の開発 遺伝環境要因の病態解明を目指すものである 2015 年 7 月に開始した三世代コホート調査は 妊婦 22,406 名 三世代家族合計 72,781 名の参加を得た (2017 年 4 月 26 日現在 ) 本研究は 分子疫学的研究による発症予測式構築 ゲノム解析による発症原因の同定 血漿プロテオーム解析 メタボローム解析による新しい早期診断バイオマーカー探索 オミックス情報の統合解析による病態メカニズムの解明を目的としている 妊娠高血圧症候群罹患群および対照群の解析データセットを確定する目的で 本コホート調査による 調査票情報 カルテ転記による臨床情報 臨床検査情報等を 順次 ToMMo 統合情報データベースに集積 しており 外れ値等の検証を含めたデータクリーニング作業を継続して実施した ToMMo の構築した統合情報データベースから 妊娠高血圧症候群罹患妊婦と夫および対照健常妊婦と夫の ID 選択 バイオバンクからの DNA 検体出庫を行い 日本人ゲノム解析に最適化したジャポニカアレイ による全ゲノム SNP スキャンを開始した また 罹患家系の HLA 型を詳細に解析するため HLA 遺伝子座 PCR プライマー設計を行った 妊娠高血圧症候群の早期発症予測バイオマーカーを探索するため 統合情報データベースから妊娠高血圧症候群罹患症例 対照例の妊娠初期血漿検体を選定し バイオバンクから出庫した 網羅性 再現性 定量性の高い SWATH 法を用い 独自開発したデータベースと連動解析することで新規バイオマーカー候補を絞り込んだ 妊娠高血圧症候群の妊婦血漿中において変動する代謝物を同定するため 解析対象症例の選定を行い 罹患症例 対照例の妊娠初期採血血漿をバイオバンクから出庫した これらの血漿を対象とした NMR によるメタボローム解析を開始すると共に MS メタボロームを一部前倒しして開始した ToMMo が構築した 日本人多層オミックス参照パネル を利活用し 病態に関連する代謝物を同定中である 当研究では 高度なセキュリティ要件を満たした ToMMo スーパーコンピュータにおいて 各研究開発 分担者が解析を行う 各研究者が共通のデータセットにアクセス可能なアカウント設定を行うことで 各 層の研究が統合的に推進可能な環境を構築した 3
4 Tohoku Medical Megabank Organization (ToMMo) launched the Birth and Three Generation Cohort Study, which is a unique cohort study with cutting-edge technologies including integrated biobank. The aim of this study is to establish the method of early prediction and elucidate undetermined pathological mechanism of hypertensive disorders of pregnancy (HDP) focused on genetic and environmental interactions. ToMMo Birth and Three Generation Cohort Study recruited 72,781 participants including 22,406 pregnant women (as of Apr 26 th, 2017). This study is composed of 1) establishment of early prediction formula (or chart) by molecular epidemiologic approaches, 2) elucidation of pathologic mechanism by whole genome analysis, 3) discovery and validation of early predictive markers by plasma proteome and metabolome, and 4) integrated bioinformatic analysis for multi-omics study. To confirm data set of HDP patients and healthy pregnant controls for epidemiologic analysis, data cleaning for outliers from information of questionnaires, clinical information by transferring medical records, and other laboratory tests was conducted. HDP affected, and control couples were selected from ToMMo integrated database. DNA samples were obtained from ToMMo biobank under satisfactory security regulations. Whole genome SNP (single nucleotide polymorphism) scanning was started using the ethnicity-specific Japonica Array which was designed by 1,070 Japanese individuals. Furthermore, PCR primers specific for each HLA genotypes were designed to analyze affected families of HDP. Plasma samples collected in the first trimester of pregnancy were obtained from ToMMo biobank to discover early predictive biomarkers of HDP. Subsequently, global proteomic analysis was performed using SWATH method with originally developed database which has noteworthy advantages for its comprehensiveness, reproducibility and quantitativity. As a result, several candidates of biomarkers were selected. To identify circulating metabolites specific for HDP, plasma samples were selected in database and obtained from ToMMo biobank. NMR metabolomics for affected and control subjects was started. Additionally, global and targeted metabolomics by mass spectrometry was accelerated ahead of research plans. Metabolites related to pathologic condition of HDP would be identified utilizing originally established, ethnicity-specific Japanese Multi-Omics Reference Panel J-Morp. This study was conducted by registered researchers using ToMMo supercomputer with highly secured requirements. Specific individual accounts for each researcher was established to access standardized formatted data set and information environment described above was enabled us to proceed integrated multi-omics analysis. 4
5 III. 成果の外部への発表 (1) 学会誌 雑誌等における論文一覧 ( 国内誌 0 件 国際誌 0 件 ) 該当ありません (2) 学会 シンポジウム等における口頭 ポスター発表該当ありません (3) 国民との科学 技術対話社会 に対する取り組み該当ありません (4) 特許出願該当ありません 5
The Tohoku Medical Megabank project is a part of the national project to reconstruct Tohoku area.. It aims to become a centripetal force for the reconstruction of Tohoku University Tohoku Medical Megabank
成果 本研究の解析で着目したのは 25 の遺伝性疾患とそれらの 57 の原因遺伝子で これらは ACMG が推奨する 偶発的 二次的所見としての遺伝情報の結果の返却を推奨する遺伝子のセットのうち常染色体上のものに相当し 大部分が遺伝性腫瘍や遺伝性循環器疾患の原因遺伝子です 本研究では 当機構で作成し
背景 全ゲノム解析や全エクソーム解析により ヒトゲノムの多様性の大量データが得られる時代となりました 当機構でも 日本人の全ゲノム解読を大規模に行い 検出された一塩基変異のリファレンスパネルを構築し 疾患と遺伝要因との関係解明のための研究に活用できるよう 国内外での利用促進を行っています 一方で ゲノム全体をカバーする解析は 疾患の原因遺伝子の塩基配列の解読もされてしまうこともあって 偶発的所見 二次的所見についての扱いのポリシーが課題になっています
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ノム解析が実施された場合に偶発的 二次的所見としての遺伝情報を知ることが患者にとって有意義としている遺伝性疾患の原因遺伝子です 具体的には 現在示されている 26 疾患 (59 遺伝子 ) のうちから 常染色体上に存在する 57 遺伝子 (25 疾患 ) を本研究の解析対象としました 2KJPN の
報道機関各位 2017 年 12 月 28 日 東北大学東北メディカル メガバンク機構 日本人一般住民が持つ疾患の原因遺伝子の変異の頻度がわかる? 日本人 2,049 人の全ゲノム解読データの調査から 発表のポイント 東北メディカル メガバンク計画で 2016 年に作成した 日本人の全ゲノムリファレンスパネル (2,049 人 ) (2KJPN) を用いて 25 の遺伝性疾患に関する 57 遺伝子
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