マイクロビーズ技術を用いた肺がん検診ツールのイノベーションと臨床応用

2016 Fiscal Year Annual Research Report

• Project/Area Number: 15H03034
• Research Institution: Okayama University
• Principal Investigator: 永坂 岳司 岡山大学, 大学病院, 講師 (30452569)
• Co-Investigator(Kenkyū-buntansha): 豊岡 伸一 岡山大学, 医歯薬学総合研究科, 教授 (30397880) ; 香川 俊輔 岡山大学, 大学病院, 准教授 (00362971) ; 楳田 祐三 岡山大学, 大学病院, 助教 (10573735) ; 母里 淑子 岡山大学, 医歯薬学総合研究科, 助教 (70708081)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 検査・診断システム / 腫瘍診断学 / 腫瘍検査学 / エピゲノム

Outline of Annual Research Achievements

本研究は喀痰に含まれる腫瘍由来Free核酸に認められる腫瘍特異的メチル化CpGを効率よく精確かつMechanicalに検出を行う技術の開発を行い，現状の肺がん検診ツールのイノベーションとその臨床応用を行うことを目的とする。平成２８年度の研究実績の概要を以下に示す。
①The Cancer Genome Atlas (TCGA) から肺がんや正常肺組織のメチル化アレイを用いたデータを参考に，喀痰からの肺がんスクリーニングに用いるマーカーの検討を行った。
②以前に収集した，合計209例の喀痰（141例が肺がん患者，68例が肺がん非担癌者）をValidationグループと定義。このValidationグループを用い，①にて得られたマーカーの精確性の検討，及び，xMAP/MAGPIXによるPCR産物内の各CpG siteのメチル化statusの詳細な検出法の最適化を行い，本スクリーニング技術の精確性の検討を行った（本研究における喀痰からの肺がん患者検出法は，Validationグループでは，AUC＝0.9４と極めて有効な値を示した）。
③呼吸器内科にて気管支内視鏡予定患者の検査施行前の喀痰の採取を開始。学内IRBにて承認を受け，呼吸器内科の協力の元，平成29年2月までに144例の気管支内視鏡患者の検査施行前の喀痰を採取。また，胸部CTにて異常所見を認めない肺がん非担癌者58例の喀痰も採取。合計202例をTESTグループと定義。②にて最適化されたxMAP/MAGPIXによるPCR産物内の各CpG siteのメチル化statusの詳細な検出法をTESTグループに用い検討を行った（TESTグループにおいても，AUC＝0.93と極めて有効な値を示し，著しい再現性を認めた）。
④これら結果をもとに，PMDAに対し，試薬・検出機器として世に出すための交渉を開始した。

Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

当初の計画以上に進展している。理由として，バイオマーカーの選択が極めてスムーズに行えたことが大きいと思われる。
また，xMAP/MAGPIXによるPCR産物内の各CpG siteのメチル化statusの詳細な検出法に関してはG&Gサイエンス社と共同開発を行っているため，確実かつ的確に最適化が行われたことも大きな要因である。

Strategy for Future Research Activity

①　PBRT PBAT(Post-bisulfite adaptor tagging)とOne Tube Methodの融合による喀痰内のゲノムワイドなメチル化領域検出法の構築（現行のマーカー群にてAUC＝0.93-0.94を確実に再現しているため，現行の方法をそのまま臨床応用に持っていく予定である。従い，PBRT PBATはもともとバイオマーカー選出がうまくいかないときの新規バイオマーカー探索のための研究であるため無理はしない）。
②　特許の取得
③　論文化
④　PMDAとの交渉継続

Research Products

• [Journal Article] Circular RNA ciRS-7 - A promising prognostic biomarker and a potential therapeutic target in colorectal cancer (2017)
  • Author(s): Weng W, Wei Q, Toden S, Yoshida K, Nagasaka T, Fujiwara T, Cai S, Qin H, Ma Y, Goel A
  • Journal Title: Clin Cancer Res. Volume: in press Pages: in press
  • DOI: 10.1158/1078-0432.CCR-16-2541
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Microsatellite Alterations With Allelic Loss at 9p24.2 Signify Less-Aggressive Colorectal Cancer Metastasis. (2016)
  • Author(s): Koi M, Garcia M, Choi C, Kim HR, Koike J, Hemmi H, Nagasaka T, Okugawa Y, Toiyama Y, Kitajima T, Imaoka H, Kusunoki M, Chen YH, Mukherjee B, Boland CR, Carethers JM.
  • Journal Title: Gastroenterology Volume: 150(4) Pages: 944-955
  • DOI: 10.1053/j.gastro.2015.12.032
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Protocol of a randomised phase III clinical trial of sequential capecitabine or 5-fluorouracil plus bevacizumab (Cape/5-FU-Bmab) to capecitabine or 5-fluorouracil plus oxaliplatin plus bevacizumab (CapeOX/mFOLFOX6-Bmab) versus combination CapeOX/mFOLFOX6-Bmab in advanced colorectal cancer: the C-cubed (C3) study. (2016)
  • Author(s): Nagasaka T, Mishima H, Sawaki A, Shimokawa M, Inukai M, Shinozaki K, Tanioka H, Nasu J, Nishina T, Hazama S, Okajima M, Yamaguchi Y.
  • Journal Title: BMJ open Volume: 6(6) Pages: e011454
  • DOI: 10.1136/bmjopen-2016-011454
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Neutrophil to lymphocyte ratio in the pre-treatment phase of final-line chemotherapy predicts the outcome of patients with recurrent ovarian cancer. (2016)
  • Author(s): Nakamura K, Nagasaka T, Nishida T, Haruma T, Ogawa C, Kusumoto T, Seki N, Hiramatsu Y.
  • Journal Title: Oncology letters Volume: 11(6) Pages: 3975-3981
  • DOI: 10.3892/ol.2016.4513
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Long-Term Survival after Radiofrequency Ablation of Lung Oligometastases from Five Types of Primary Lesions: A Retrospective Evaluation. (2016)
  • Author(s): Omae K, Hiraki T, Gobara H, Iguchi T, Fujiwara H, Matsui Y, Toyooka S, Nagasaka T, Kanazawa S.
  • Journal Title: Journal of vascular and interventional radiology : JVIR Volume: 27(9) Pages: 1362-1370
  • DOI: 10.1016/j.jvir.2016.05.017
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Risk Factors for the Development of Desmoid Tumor After Colectomy in Patients with Familial Adenomatous Polyposis: Multicenter Retrospective Cohort Study in Japan. (2016)
  • Author(s): Saito Y, Hinoi T, Ueno H, Kobayashi H, Konishi T, Ishida F, Yamaguchi T, Inoue Y, Kanemitsu Y, Tomita N, Matsubara N, Komori K, Kotake K, Nagasaka T, Hasegawa H, Koyama M, Ohdan H, Watanabe T, Sugihara K, Ishida H.
  • Journal Title: Annals of surgical oncology Volume: 23 Pages: 559-565
  • DOI: 10.1245/s10434-016-5380-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Single Center Prospective Phase II Trial of CT-guided Radiofrequency Ablation for Pulmonary Metastases from Colorectal Cancer (SCIRO-1401). (2016)
  • Author(s): Sakurai J, Matsui Y, Hiraki T, Iguchi T, Fujiwara H, Gobara H, Mitsuhashi T, Nagasaka T, Kanazawa S.
  • Journal Title: Acta medica Okayama Volume: 70(4) Pages: 317-321
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Anti-high mobility group box 1 monoclonal antibody improves ischemia/reperfusion injury and mode of liver regeneration after partial hepatectomy. (2016)
  • Author(s): Sugihara M, Sadamori H, Nishibori M, Sato Y, Tazawa H, Shinoura S, Umeda Y, Yoshida R, Nobuoka D, Utsumi M, Ohno K, Nagasaka T, Yoshino T, Takahashi HK, Yagi T, Fujiwara T.
  • Journal Title: American journal of surgery Volume: 211(1) Pages: 179-188
  • DOI: 10.1016/j.amjsurg.2015.06.025
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prevalence of laparoscopic surgical treatment and its clinical outcomes in patients with familial adenomatous polyposis in Japan (2016)
  • Author(s): Ueno H, Kobayashi H, Konishi T, Ishida F, Yamaguchi T, Hinoi T, Kanemitsu Y, Inoue Y, Tomita N, Matsubara N, Komori K, Ozawa H, Nagasaka T, Hasegawa H, Koyama M, Akagi Y, Yatsuoka T, Kumamoto K, Kurachi K, Tanakaya K, Yoshimatsu K, Watanabe T, Sugihara K, Ishida H.
  • Journal Title: International journal of clinical oncology Volume: 21(4) Pages: 713-722
  • DOI: 10.1007/s10147-016-0953-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Expansion of epigenetic alterations in EFEMP1 promoter predicts malignant formation in pancreatobiliary intraductal papillary mucinous neoplasms. (2016)
  • Author(s): Yoshida K, Nagasaka T, Umeda Y, Tanaka T, Kimura K, Taniguchi F, Fuji T, Shigeyasu K, Mori Y, Yanai H, Yagi T, Goel A, Fujiwara T.
  • Journal Title: Journal of cancer research and clinical oncology Volume: 142(7) Pages: 1557-1569
  • DOI: 10.1007/s00432-016-2164-x
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Feasibility of sequential adjuvant chemotherapy with a 3-month oxaliplatin-based regimen followed by 3 months of capecitabine in patients with stage III and high-risk stage II colorectal cancer: JSWOG-C2 study. (2016)
  • Author(s): Tsuruta A, Yamashita K, Tanioka H, Tsuji A, Inukai M, Yamakawa T, Yamatsuji T, Yoshimitsu M, Toyota K, Yamano T, Nagasaka T, Okajima M.
  • Journal Title: Drug design, development and therapy Volume: 10 Pages: 3827-3835
  • DOI: 10.2147/DDDT.S112322
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Novel assay for the detection of methylated CpGs from sputum to screen patients with lung cancer (2016)
  • Author(s): Takeshi Nagasaka, Akihiro N, Toshiaki T, Takeshi K, Kazuya Y, Keisuke K, Yoshiko M, Hiroyuki K, Toshiyoshi F
  • Organizer: ESMO Asia 2016
  • Place of Presentation: Singapore
  • Year and Date: 2016-12-16 – 2016-12-19
  • Int'l Joint Research
• [Presentation] 遺伝子変異情報を考慮したStageIV大腸癌に対する予後の検討及び治療戦略の構築 (2016)
  • Author(s): 永坂岳司， 母里淑子， 岸本浩行， 神原健， 河合毅， 中谷紳， 嶋村廣視， 富岡憲明， 森 谷行利， 瀧上隆夫， 佃和憲， 藤原俊義
  • Organizer: 第71回日本大腸肛門病学会学術集会
  • Place of Presentation: 伊勢
  • Year and Date: 2016-11-18 – 2016-11-19
• [Presentation] 当院におけるmFOLFOXIRIの検討 (2016)
  • Author(s): 母里淑子，永坂岳司， 岸本浩行， 河合毅， 佃和憲， 藤原俊義
  • Organizer: 第71回日本大腸肛門病学会学術集会
  • Place of Presentation: 伊勢
  • Year and Date: 2016-11-18 – 2016-11-19
• [Presentation] リンチ症候群における遺伝子変異の特徴とスクリーニング， PD-1 を含めた治療標的 (2016)
  • Author(s): 永坂岳司, 谷口文崇, 母里淑子, 藤原俊義
  • Organizer: 第14回日本臨床腫瘍学会学術集会
  • Place of Presentation: 神戸
  • Year and Date: 2016-07-28 – 2016-07-30
• [Presentation] ランチョンセミナーI 大腸癌の術後補助化学療法 (2016)
  • Author(s): 永坂岳司
  • Organizer: 第25回日本がん転移学会学術集会・総会
  • Place of Presentation: 米子
  • Year and Date: 2016-07-21 – 2016-07-22
  • Invited
• [Presentation] Acquired Resistance to Anti-Epidermal Growth Factor Receptor Therapy in Metastatic Colorectal Cancer (2016)
  • Author(s): 谷口文崇, 永坂岳司, 藤 智和, 母里淑子, 岸本浩行, 河合 毅, 楳田祐三, 藤原俊義
  • Organizer: 第71回日本消化器外科学会総会
  • Place of Presentation: 徳島
  • Year and Date: 2016-07-14 – 2016-07-16
• [Presentation] Lynch症候群および家族性大腸腺腫症の系統的遺伝子解析 (2016)
  • Author(s): 谷口文崇, 永坂岳司, 楳田祐三, 母里淑子, 河合 毅, 内海方嗣, 田中屋宏爾, 青木秀樹, 竹内仁司, 藤原俊義
  • Organizer: 第85回大腸癌研究会
  • Place of Presentation: 大阪
  • Year and Date: 2016-07-01 – 2016-07-01
• [Presentation] 4つのモノヌクレオチドリピートマーカーのミスマッチ修復タンパク欠損を伴う大腸癌・子宮体癌スクリーニングの精確性の検討 (2016)
  • Author(s): 母里淑子, 永坂岳司, 谷口文崇, 河合 毅, 岸本浩行, 神原 健, 藤原俊義
  • Organizer: 第85回大腸癌研究会
  • Place of Presentation: 大阪
  • Year and Date: 2016-07-01 – 2016-07-01
• [Presentation] 切除不能進行・再発大腸癌治療におけるBest Practice －抗EGFR抗体薬の使い方－ (2016)
  • Author(s): 永坂岳司
  • Organizer: 第4回岡山大腸腫瘍研究会
  • Place of Presentation: 岡山
  • Year and Date: 2016-06-14 – 2016-06-14
  • Invited
• [Presentation] 4つのモノヌクレオチドリピートマーカーを用いたマイクロサテライト不安定性腫瘍検出方法の検討 (2016)
  • Author(s): 永坂岳司, 竹原裕子, 入谷光洋, 母里淑子, 春間朋子, 原賀順子, 中村圭一郎, 平松祐司, 藤原俊義
  • Organizer: 第22回日本家族性腫瘍学会学術集会
  • Place of Presentation: 松山
  • Year and Date: 2016-06-03 – 2016-06-04
• [Presentation] MGMT Inactivation Arrest Tumor Growth and Serves As a Promising Predictive Biomarker for Treatment Response in Colorectal Cancer (2016)
  • Author(s): Mori Y, Nagasaka T, Tazawa H, Umeda Y, Shigeyasu K, Yoshida K, Goel A, Fujiwara T
  • Organizer: Digestive Disease Week (DDW) 2016
  • Place of Presentation: San Diego
  • Year and Date: 2016-05-21 – 2016-05-24
  • Int'l Joint Research
• [Presentation] A novel circulating cell free DNA-based assay in colorectal cancer patients during treatment with systematic chemotherapy (2016)
  • Author(s): Toshima T, Nagasaka T, Mori Y, Kawai T, Fuji T, Taniguchi F, Kimura K, Yasui K, Kishimoto H, Umeda Y, Tazawa H, Goel A, Fujiwara T
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans20
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] PD-1 and PD-L1 expression patterns and DNA mismatch repair status for precision management of patients with gastric cancer (2016)
  • Author(s): Kimura K, Nagasaka T, Mori Y, Kawai T, Fuji T, Taniguchi F, Yasui K, Toshima T, Umeda Y, Tazawa H, Goel A, Fujiwara T
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] Micro RNAs as promising therapeutic targets for anti-metastatic therapy in colorectal cancer (2016)
  • Author(s): Kawai T, Nagasaka T, Mori Y, Fuji T, Taniguchi F, Kimura K, Toshima T, Yasui K, Umeda Y, Tazawa H, Goel A, Fujiwara T
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] Genome-wide enhancer analysis identifies PVT1 as an oncogenic enhancer of MYC (2016)
  • Author(s): Shigeyasu K, Toden S, Nagasaka T, Fujiwara T, Boland CR, Goel A
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] A novel live imaging system for inflammation-induced epithelial-mesenchymal transition in colorectal cancers (2016)
  • Author(s): eda T, Tazawa H, Kikuchi S, Kuroda S, Ohara T, Noma K, Kishimoto H, Nagasaka T, Nishizaki M, Kagawa S, Fujiwara T
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] Comprehensive analysis of intrahepatic cholangiocarcinoma based on viral infections and mutational status in the IDH1/2 and KRAS genes (2016)
  • Author(s): Yasui K, Nagasaka T, Umeda Y, Fuji T, Taniguchi F, Toshima T, Kimura K, Kawai T, Mori Y, Tazawa H, Yagi T, Goel A, Fujiwara T
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] Clinical significance of SNORA21, an H/ACA box snoRNA, as a metastasis predicting and prognostic biomarker in colorectal cancer (2016)
  • Author(s): Yoshida K, Toden S, Weng W, Nagasaka T, Fujiwara T, Goel A
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] 切除可能局所進行直腸癌に対する術前化学療法および術前放射線化学療法の検討 (2016)
  • Author(s): 母里淑子, 永坂岳司, 岸本浩行, 河合 毅, 工藤泰崇, 樹下真希, 近藤喜太, 浅野博昭, 佃 和憲, 藤原俊義
  • Organizer: 第116回日本外科学会定期学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2016-04-14 – 2016-04-16
• [Presentation] 膵癌切除症例における免疫チェックポイント分子の発現解析～膵癌切除例とTCGA dataを対象とした免疫チェックポイント機構の解析～ (2016)
  • Author(s): 藤 智和, 楳田祐三, 永坂岳司, 母里淑子, 谷口文崇, 安井和也, 河合 毅, 八木孝仁, 藤原俊義
  • Organizer: 第116回日本外科学会定期学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2016-04-14 – 2016-04-16
• [Presentation] Clinical Significance of MLH1 Methylation and CpG Island Methylator Phenotype as Prognostic Markers in Patients with Gastric Cancer (2016)
  • Author(s): Mori Y, Shigeyasu K, Nagasaka T, ToyookaS, Fujiwara T
  • Organizer: ICHG2016 第13回国際人類遺伝学会
  • Place of Presentation: 京都
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Genetic and epigenetic alterations of netrin-1 receptors in gastric cancer (2016)
  • Author(s): Nagasaka T, Mori Y, Shigeyasu K, Toyooka S, Fujiwara T
  • Organizer: ICHG2016 第13回国際人類遺伝学会
  • Place of Presentation: 京都
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research