Gene analysis in melanoma patients using circulating tumor DNA

• Project/Area Number: 26461687
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Dermatology
• Research Institution: Shinshu University
• Principal Investigator: ASHIDA Atsuko 信州大学, 学術研究院医学系, 助教 (00596786)
• Co-Investigator(Kenkyū-buntansha): 宇原 久 札幌医科大学, 医学部, 教授 (40201355) ; 奥山 隆平 信州大学, 学術研究院医学系, 教授 (80292332)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ctDNA / メラノーマ / BRAF / NRAS / バイオマーカー / circulating tumor DNA

Outline of Final Research Achievements

Circulating tumor DNA (ctDNA) is a novel biomarker for evaluating the features of　tumors in cancer patients. Detection of ctDNA usually requires specialized equipment, because blood includes only a small amount of the tumor- derived DNA. We assessed the utility of BRAF- and NRAS mutated ctDNA levels using competitive allele specific TaqMan PCR (castPCR) or droplet digital PCR. ctDNA was detected in cases with heavy tumor burden as advanced disease. Detection of ctDNA was associated with poor prognosis. Quantification of ctDNA (BRAFmutant or NRASmutant) levels during the treatment course (nivolumab or vemurafenib) showed dynamic changes corresponding to radiologic and clinical alterations. Our finding suggest that ctDNA might be a useful biomarker of clinical response and tumor recurrence while monitoring patients with metastatic melanoma.

Research Products

• [Journal Article] Detection of BRAFV600K mutant tumor-derived DNA in the pleural effusion from a patient with metastatic melanoma (2017)
  • Author(s): Sakaizawa K, Ashida A, Uhara H, Okuyama R.
  • Journal Title: Clin Chem Lab Med Volume: 55 Issue: 4 Pages: 92-95
  • DOI: 10.1515/cclm-2016-0634
  • Peer Reviewed / Open Access
• [Journal Article] Melanoma with BRAF Mutation in Circulating Cell-free DNA despite no Mutation in the Primary Lesion: A Case Report (2016)
  • Author(s): Atsuko Ashida, Hisashi Uhara, Asuka Mikoshiba, Kaori Sakaizawa, Naomi Kumagai, Hiroshi Koga and Ryuhei Okuyama
  • Journal Title: Acta Derm Venereol Volume: 96 Issue: 1 Pages: 128-129
  • DOI: 10.2340/00015555-2194
  • NAID: 120007100151
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Quantitative analysis of the BRAFV600E mutation in circulating tumor-derived DNA using competitive allele-specific TaqMan PCR in melanoma patients (2016)
  • Author(s): Ashida A, Sakaizawa K, Mikoshiba A, Uhara H, Okuyama R
  • Journal Title: Int J Clin Oncol Volume: 21 Issue: 5 Pages: 981-988
  • DOI: 10.1007/s10147-016-0976-y
  • Peer Reviewed / Open Access
• [Journal Article] Acquired epidermodysplasia verruciformis associated with human papillomavirus type 47 in a HIV-infected patient (2016)
  • Author(s): Atsuko Ashida, Akira Shimizu, Ryuhei Okuyama
  • Journal Title: J Dermatol Volume: 印刷中 Issue: 6 Pages: 53-56
  • DOI: 10.1111/1346-8138.13262
  • Peer Reviewed / Open Access
• [Journal Article] Inhibition of epidermal growth factor receptor sinduces tumor necrosis factor-a via activation of peroxisome proliferator-activated receptor-g and nuclear factor-kB in sebocytes: A possible pathogenesis of papulopustular rash (2016)
  • Author(s): Atsuko Ashida, Eisaku Ogawa,, Hisashi Uhara, Tomonobu Koizumi, Ryuhei Okuyama
  • Journal Title: J Dermatol Sci Volume: 82 Issue: 1 Pages: 53-56
  • DOI: 10.1016/j.jdermsci.2015.12.004
  • Peer Reviewed / Open Access
• [Journal Article] An Initial Case Report of a Laparoscopic Spleen-Preserving Distal Pancreatectomy for a Patient with a Pancreatic Metastasis of Malignant Melanoma (2015)
  • Author(s): Motoyama H, Kobayashi A, Yokoyama T, Shimizu A, Furusawa N , Sakai H, Kitagawa N, Ashida A, Okuyama R, Miyagawa S
  • Journal Title: 信州医誌 Volume: 63 Pages: 19-24
  • Peer Reviewed
• [Journal Article] Clinical characteristics associated with BRAF, NRAS and KIT mutations in Japanese melanoma patients. (2015)
  • Author(s): Sakaizawa K, Ashida A, Uchiyama A, Ito T, Fujisawa Y, Ogata D, Matsushita S, Fujii K, Fukushima S, Shibayama Y, Hatta N, Takenouchi T, Uehara J, Okuyama R, Yamazaki N, Uhara H.
  • Journal Title: J Dermatol Sci. Volume: 80 Issue: 1 Pages: 33-37
  • DOI: 10.1016/j.jdermsci.2015.07.012
  • Peer Reviewed / Open Access
• [Journal Article] Histopathological improvement of scleroderma induced by paclitaxel in a patient with breast cancer (2015)
  • Author(s): Ohashi A, Minagawa A, Ashida A, Koga H, Uhara H, Okuyama R
  • Journal Title: J Dermatol Volume: 42 Issue: 12 Pages: 1198-1199
  • DOI: 10.1111/1346-8138.13085
  • Peer Reviewed / Open Access
• [Journal Article] NRAS mutations in primary and metastatic melanomas of Japanese patients (2013)
  • Author(s): Uhara H, Ashida A, Koga H, Uchiyama A, Uchiyama R, Hayashi K, Kiniwa Y, Okuyama R
  • Journal Title: Int J Clin Oncol Volume: - Issue: 3 Pages: 544-548
  • DOI: 10.1007/s10147-013-0573-2
  • Peer Reviewed
• [Presentation] 末梢血中の腫瘍由来DNAを用いた悪性黒色腫に対するニボルマブ治療効果の評価 (2017)
  • Author(s): 芦田敦子、境澤香里、宇原久、奥山隆平
  • Organizer: 第193回日本皮膚科学会信州地方会
  • Place of Presentation: 松本市（信州大学）
  • Year and Date: 2017-03-27
• [Presentation] Quantitative analysis of the BRAFV600E in circulating tumor-drived DNA using competitive allele-specific TaqMan PCR in melanoma patients (2016)
  • Author(s): 芦田敦子、境澤香里、宇原久、奥山隆平
  • Organizer: 第30回表皮細胞研究会
  • Place of Presentation: 弘前市（アートホテル弘前シティ）
  • Year and Date: 2016-11-12
• [Presentation] 胸水中のcell-free DNAよりBRAFV600Kを検出できた進行期悪性黒色腫の１例 (2016)
  • Author(s): 境澤香里、芦田敦子、宇原久、奥山隆平
  • Organizer: 第80回日本皮膚科学会東部支部学術大会
  • Place of Presentation: 浜松市（アクトシティ浜松）
  • Year and Date: 2016-10-29
• [Presentation] Evaluation of response to nivolumab treatment in patients with metastatic melanoma using circulating tumor DNA (2016)
  • Author(s): 芦田敦子、境澤香里、宇原久、奥山隆平
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: 横浜市（パシフィコ横浜）
  • Year and Date: 2016-10-08
• [Presentation] 血液循環腫瘍DNAよりBRAF変異を検出した進行期悪性黒色腫の１例 (2016)
  • Author(s): 芦田敦子、御子柴飛鳥、境澤香里、古賀弘志、熊谷尚美、宇原久、奥山隆平
  • Organizer: 第115回日本皮膚科学会学術総会
  • Place of Presentation: 京都市（国立京都国際会館）
  • Year and Date: 2016-06-03
• [Presentation] メラノーマの新しい治療法「メラノーマの遺伝子異常」 (2016)
  • Author(s): 芦田敦子
  • Organizer: 第115回日本皮膚科学会学術総会
  • Place of Presentation: 京都市（国立京都国際会館）
  • Year and Date: 2016-06-03
  • Invited
• [Presentation] 血液循環腫瘍DNAよりBRAF変異を検出した進行期悪性黒色腫症例 (2016)
  • Author(s): 芦田敦子、御子柴飛鳥、境澤香里、古賀弘志、熊谷尚美、宇原久、奥山隆平
  • Organizer: 第115回日本皮膚科学会総会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2016-06-03
• [Presentation] HIV感染患者に生じた後天性疣贅状表皮発育異常症の一例 (2016)
  • Author(s): 芦田敦子、大橋敦子、清水晶、奥山隆平
  • Organizer: 第79回東京・東部支部合同学術大会
  • Place of Presentation: 京王プラザホテル（東京都新宿区）
  • Year and Date: 2016-02-20
• [Presentation] Epidermal growth factor receptor inhibitor induces tumor necrosis factor-alpha; via activation of the PPARgamma; and NF-kB from sebocytes (2015)
  • Author(s): Atsuko Ashida, Eisaku Ogawa, Ryuhei Uchiyama, Hisashi Uhara, Ryuhei Okuyama
  • Organizer: 第40回日本研究皮膚科学会学術大会総会
  • Place of Presentation: 岡山コンベンションセンター（岡山市）
  • Year and Date: 2015-12-08
• [Presentation] Detection of BRAFV600Ecirculating tumomr DNA using castPCR in patients with advanced melanoma (2015)
  • Author(s): 第114回日本皮膚科学会総会
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場（名古屋市）
  • Year and Date: 2015-10-08
• [Presentation] EGFR阻害薬による皮膚障害とその対策・予防 (2015)
  • Author(s): 芦田敦子、高沢裕子、関智子、境澤香里、羽生田久美子、上條文史尚、皆川茜、奥山隆平
  • Organizer: 第188回信州地方会
  • Place of Presentation: 信州大学（長野県松本市）
  • Year and Date: 2015-07-14
• [Presentation] 日本人の悪性黒色腫のBRAF, NRAS, KIT変異率と臨床所見 (2015)
  • Author(s): 境澤香里、芦田敦子、伊東孝通、藤澤康弘、緒方大、松下茂人、藤井一恭、福島聡、柴山慶継、八田尚人、竹之内辰也、上原治朗、奥山隆平、山﨑直也、宇原久
  • Organizer: 第114回日本皮膚科学会総会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2015-05-29
• [Presentation] 皮膚癌の術後出血が診断の契機となった後天性13因子欠乏症の１例 (2015)
  • Author(s): 三宅知美、芦田敦子、宇原久、奥山隆平、福澤正男
  • Organizer: 第114回日本皮膚科学会総会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2015-05-29
• [Book] カレントテラピー「メラノーマ基礎から最新薬物療法まで」 (2016)
  • Author(s): 芦田敦子、木庭幸子、宇原久
  • Total Pages: 90
  • Publisher: ライフメディコム
• [Book] メラノーマー基礎から最新薬物治療までー 分子異常 (2016)
  • Author(s): 芦田敦子、木庭幸子、宇原久
  • Publisher: カレントテラピー
• [Book] 家族性腫瘍ー家族性腫瘍の最新研究動向ーFamilial atypical multiple mole melanoma syndrome (2015)
  • Author(s): 芦田敦子、宇原久
  • Publisher: 日本臨床
• [Book] Derma メラノーマ最新情報「日本人におけるMAPK系の遺伝子変異率と遺伝子検査の実際」 (2015)
  • Author(s): 芦田敦子、宇原久
  • Total Pages: 76
  • Publisher: 全日本病院出版会
• [Book] 家族性腫瘍 (2015)
  • Author(s): 芦田敦子、宇原久
  • Publisher: 日本臨床
• [Book] 皮膚悪性腫瘍 悪性黒色腫 「分子標的薬 KITを標的とした治療薬」 (2014)
  • Author(s): 芦田敦子、奥山隆平
  • Total Pages: 364
  • Publisher: 中山書店