Development of non-invasive diagnostic tools for childhood cancers using digital PCR

• Project/Area Number: 26861483
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Pediatric surgery
• Research Institution: Hiroshima University
• Principal Investigator: KURIHARA SHO 広島大学, 大学病院, 医科診療医 (40724894)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 癌 / 遺伝子 / 小児

Outline of Final Research Achievements

MYCN amplification was detected by digital PCR using of plasma DNA and bone marrow DNA samples derived from 23 neuroblastoma cases before treatment but became undetectable after surgery except for 6 cases with metastasis. ALK mutations in 2 neuroblastoma cases and CTNNB1 mutations in 3 hepatoblastoma cases were detected in plasma DNA at diagnosis but became undetectable more than 5 days after surgery. The library for 12 kinds of chimeric DNAs in small round cell tumors was designed. By the next generation sequencing using this library, chimeric genes were detected in 1 Ewing family tumor and 2 rhabdomyosarcomas but became undetecble after surgery. Plasma exosomes were concentrated and then microRNA was detected in these exosome samples in 4 cases.
These data suggested that digital PCR methods using blood sample is useful for diagnosis of childhood cancers.

Research Products

• [Journal Article] Circulating free DNA as non-invasive diagnostic biomarker for childhood solid tumors (2015)
  • Author(s): Kurihara S, Ueda Y, Onitake Y, Sueda T, Ohta E, Morihara N, Hirano S, Irisuna F, Hiyama E
  • Journal Title: Journal of Pediatric Surgery Volume: 50 Issue: 12 Pages: 2094-2097
  • DOI: 10.1016/j.jpedsurg.2015.08.033
  • Peer Reviewed / Open Access
• [Journal Article] Mortality and morbidity in primarily resected hepatoblastomas in Japan: Experience of the JPLT (Japanese Study Group for Pediatric Liver Tumor) trials (2015)
  • Author(s): Hiyama E, Hishiki T, Watanabe K, Ida K, Yano M, Oue T, Iehara T, Hoshino K, Koh K, Tanaka Y, Kurihara S, Ueda Y, Onitake Y
  • Journal Title: Journal of Pediatric Surgery Volume: 50 Issue: 12 Pages: 2098-2101
  • DOI: 10.1016/j.jpedsurg.2015.08.035
  • Peer Reviewed / Open Access
• [Journal Article] 肝芽腫の分子生物学 (2015)
  • Author(s): 檜山英三, 上田祐華, 栗原 将
  • Journal Title: 小児外科 Volume: 47 Pages: 181-188
  • Peer Reviewed
• [Journal Article] Clinical features of ATRX or DAXX mutated neuroblastoma (2014)
  • Author(s): Kurihara S, Hiyama E, Onitake Y, Yamaoka E, Hiyama K
  • Journal Title: Journal of Pediatric Surgery Volume: 49 Issue: 12 Pages: 1835-1838
  • DOI: 10.1016/j.jpedsurg.2014.09.029
  • Peer Reviewed / Open Access
• [Journal Article] Pediatric Hepatoblastoma: diagnosis and treatment (2014)
  • Author(s): Hiyama E
  • Journal Title: Translational Pediatrics Volume: 3 Pages: 293-299
  • Peer Reviewed / Open Access
• [Presentation] 門脈体循環シャントにて経過観察している3症例 (2015)
  • Author(s): 栗原 将, 檜山 英三, 上田 祐華
  • Organizer: 第31回日本小児外科学会秋季シンポジウム
  • Place of Presentation: 熊本市
  • Year and Date: 2015-10-31
• [Presentation] Prediction of prognosis by preoperative chemotherapy response in hepatoblastoma patients treated by JPLT-2 protocol (2015)
  • Author(s): Hiyama E, Hishiki T, Ida K, Watanabe K, Oue T, Yano M, Hoshino K, Iehara T, Koh K, Tanaka Y, Kurihara S, Ueda Y
  • Organizer: SIOP 2015
  • Place of Presentation: Cape Town, South Africa
  • Year and Date: 2015-10-08
  • Int'l Joint Research
• [Presentation] Integrated exome analysis in childhood hepatoblastoma: Biological approach for molecular targetinｇ (2015)
  • Author(s): Hiyama E, Kurihara S, Onitake Y, Ueda Y, Morihara N, Fukuba I, Komatsu R
  • Organizer: SIOP 2015
  • Place of Presentation: Cape Town, South Africa
  • Year and Date: 2015-10-08
  • Int'l Joint Research
• [Presentation] AFP高値を認めた肝限局性過形成の1症例 (2015)
  • Author(s): 栗原 将
  • Organizer: 第54回日本小児外科学会中国四国地方会
  • Place of Presentation: 愛媛県松山市
  • Year and Date: 2015-09-26
  • Int'l Joint Research
• [Presentation] 門脈－右房シャントを呈した静脈管欠損症の1例 (2015)
  • Author(s): 栗原 將, 檜山 英三, 鬼武 美幸
  • Organizer: 第52回日本小児外科学会学術集会
  • Place of Presentation: 兵庫県神戸市
  • Year and Date: 2015-05-28
• [Presentation] Circulating free DNA as non-invasive diagnostic biomarker for childhood solid tumors (2015)
  • Author(s): Kurihara S, Ueda Y, Onitake Y, Sueda T, Ohta E, Morihara N, Hirano S, Irisuna F, Hiyama E
  • Organizer: 48th Annual Meeting of the Pacific Association of Pediatric Surgeons
  • Place of Presentation: Jeju, South Kore
  • Year and Date: 2015-05-17
  • Int'l Joint Research
• [Presentation] Mortality and morbidity in primarily resected hepatoblastomas in Japan: experience of JPLT (Japanese study group for pediatric liver tumor) trials (2015)
  • Author(s): Hiyama E, Hishiki T, Watanabe K, Ida K, Yano M, Oue T, Iehara T, Hoshino K, Koh K, Tanaka Y, Kurihara S, Ueda Y, Onitake Y
  • Organizer: 48th Annual Meeting of the Pacific Association of Pediatric Surgeons
  • Place of Presentation: Jeju, South Kore
  • Year and Date: 2015-05-17
  • Int'l Joint Research
• [Presentation] 肺転移を有した肝芽腫6例の外科的検討 (2014)
  • Author(s): 栗原 将, 鬼武美幸, 三木瑞香, 中村和洋, 小林正夫, 檜山英三
  • Organizer: 第56回日本小児血液・がん学会学術集会
  • Place of Presentation: 岡山市
  • Year and Date: 2014-11-28 – 2014-11-30
• [Presentation] Outcome and morbidity of primary resection of hepatoblastoma in JPLT-1 and 2 protocols (2014)
  • Author(s): Hiyama E, Hishiki T, Watanabe K, Ida K, Yano M, Oue T, Iehara T, Hoshino K, Koh K, Tanaka Y, Kurihara S
  • Organizer: SIOP2014
  • Place of Presentation: Toronto, Canada
  • Year and Date: 2014-10-22 – 2014-10-25
• [Presentation] 小児固形腫瘍における循環遊離DNAと循環がん細胞のがん診断への応用 (2014)
  • Author(s): 檜山英三, 檜山桂子, 林 陽子, 森原なぎさ, 平野尚子, 福場郁子, 山岡絵美, 栗原 将
  • Organizer: 第73回日本癌学会学術集会
  • Place of Presentation: 横浜市
  • Year and Date: 2014-09-27
• [Presentation] urgical managements for refractory neuroblastoma in young children (2014)
  • Author(s): Hiyama E, Kurihara S, Onitake Y, Ueda Y, Miki M, Kawaguchi H, Nakamura K, Kobayashi M
  • Organizer: BAPS61st Annual Congress
  • Place of Presentation: Edinburgh, Scotland
  • Year and Date: 2014-07-22 – 2014-07-25
• [Presentation] Central venous catheter-related complications in children with malignancy (2014)
  • Author(s): Kurihara S, Hiyama E, Onitake Y, Miki M, Nakamura K, Kawaguchi H, Kobayashi M
  • Organizer: PAPS2014
  • Place of Presentation: Calgary, Canada
  • Year and Date: 2014-05-25 – 2014-05-29
• [Presentation] Clinical feature of ATRX or DAXX mutated neuroblastoma (2014)
  • Author(s): Hiyama E, Kurihara S, Onitake Y, Yamaoka E, Fukuba I, Hiyama K
  • Organizer: PAPS2014
  • Place of Presentation: Calgary, Canada
  • Year and Date: 2014-05-25 – 2014-05-29
• [Presentation] 肝芽腫の縦隔再発に対してICG蛍光法を使用した経験 (2014)
  • Author(s): 栗原 将, 鬼武美幸, 小倉 薫, 檜山英三
  • Organizer: 第55回日本中国四国小児がん研究会
  • Place of Presentation: 広島県福山市
  • Year and Date: 2014-04-26