Gene analysis in circulating melanoma cells isolated using novel monoclonal antibodies

• Project/Area Number: 23591616
• 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, 医学部, 助教 (00596786)
• Co-Investigator(Kenkyū-buntansha): GOTO Yasufumi 信州大学, 医学部, 非常勤講師 (60467181) ; OKUYAMA Ryuhei 信州大学, 医学部, 教授 (80292332)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2011: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 血液循環腫瘍細胞 / メラノーマ / B7-H3 / CTCs

Research Abstract

We investigated an optimized procedure for the efficient isolation of melanoma cells labeled with HMW-MAA (High-Molecular Weight Melanoma-Associated Antigen) -specific antibodies and/or B7-H3-specific antibodies from the peripheral blood using immunomagnetic beads and fluorescence-activated cell sorting (FACS). Compared with the single-labeling method of melanoma cells using HMW-MAA or B7-H3 antibodies, the double-labeling method using HMW-MAA and B7-H3 antibodies earned high yield.These results suggest that the double-labeling method using HMW-MAA and B7-H3 antibodies is useful for isolating melanoma cells from peripheral blood in patients with melanoma.

Research Products

• [Journal Article] NRAS mutations in primary and metastatic melanomas of Japanese patients (2014)
  • 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: (in press)
  • Peer Reviewed
• [Journal Article] 5-hydroxymethylcytosine as a useful marker to differentiate between malignant melanomas and benign melanocytic nevi (2014)
  • Author(s): Uchiyama R, Uhara H, Uchiyama A, Ogawa E, Takazawa Y, Ashida A, Koga H, Hayashi K, Kiniwa Y, Okuyama R
  • Journal Title: J Dermatol Sci Volume: 73 Pages: 161-163
  • Peer Reviewed
• [Journal Article] Successful treatment of rheumatoid vasculitis-associated skin ulcer with a TNFα antagonist (2014)
  • Author(s): Ashida A, Murata H, Mikoshiba Y, Ohashi A, Kobayashi A, Koga H, Uhara H, Okuyama R
  • Journal Title: Int J Dermatol Volume: 53 Issue: 2
  • DOI: 10.1111/j.1365-4632.2012.05529.x
  • Peer Reviewed
• [Journal Article] Combination chemotherapy of carboplatin and paclitaxel for metastatic melanoma (2013)
  • Author(s): Sato Y, Uhara H, Ashida A, Okuyama R
  • Journal Title: J Dermatol Volume: 40 Pages: 1050-1051
  • Peer Reviewed
• [Journal Article] チロシンキナーゼ:KIT (2013)
  • Author(s): 芦田敦子、木庭幸子、宇原久、奥山隆平
  • Journal Title: 日本臨床 Volume: 71増刊号4 Pages: 134-138
• [Journal Article] MAPキナーゼ:NRAS、BRAF、MEK (2013)
  • Author(s): 芦田敦子、木庭幸子、宇原久、奥山隆平
  • Journal Title: 日本臨床 Volume: 71増刊号4 Pages: 126-129
• [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
• [Journal Article] チロシンキナーゼ：KIT (2013)
  • Author(s): 芦田敦子、木庭幸子、宇原久、奥山隆平
  • Journal Title: 日本臨床 Volume: 増刊号4 Pages: 134-138
• [Journal Article] MAPキナーゼ：NRAS、BRAF、MEK (2013)
  • Author(s): 芦田敦子、木庭幸子、宇原久、奥山隆平
  • Journal Title: MAPキナーゼ：NRAS、BRAF、MEK Volume: 増刊号4 Pages: 126-129
• [Journal Article] Low p53 positivity in verrucous skin lesion in diabetic neuropathy occurring on the dorsum of the foot. (2013)
  • Author(s): Ashida A, Kiniwa Y, Kobayashi A, Matsumoto K, Okuyama R.
  • Journal Title: Int J Dermatol Volume: 52 Issue: 3 Pages: 378-380
  • DOI: 10.1111/j.1365-4632.2011.04891.x
  • Peer Reviewed
• [Journal Article] A case of hypocomplementemic urticarial vasculitis with a high serum level of rheumatoid factor (2013)
  • Author(s): Ashida A, Murata H, Ohashi A, Ogawa E, Uhara H, Okuyama R
  • Journal Title: Australas J Dermatol Volume: 54 Issue: 3 Pages: e62-e63
  • DOI: 10.1111/j.1440-0960.2011.00873.x
  • Peer Reviewed
• [Journal Article] 皮膚悪性腫瘍 基礎と臨床の最新研究動向 MAPキナーゼ経路:NRAS,BRAF,MEK (2013)
  • Author(s): 芦田敦子
  • Journal Title: 日本臨床 Volume: in press
• [Journal Article] 皮膚悪性腫瘍 基礎と臨床の最新研究動向 チロシンキナーゼ：KIT (2013)
  • Author(s): 芦田敦子
  • Journal Title: 日本臨床 Volume: in press
• [Journal Article] Assessment of BRAF and KIT mutations in Japanese melanoma patients (2012)
  • Author(s): Ashida A, Uhara H, Kiniwa Y, Oguchi M, Murata H, Goto Y, Uchiyama A, Ogawa E, Hayashi K, Koga H, Okuyama R
  • Journal Title: J Dermatol Sci Volume: 66 Pages: 240-242
  • Peer Reviewed
• [Journal Article] Mutation analysis of BRAF and KIT in circulating melanoma cells at the single cell level (2012)
  • Author(s): SakaizawaK, Goto Y, Kiniwa Y, Uchiyama A, Harada K, shimada S, Saida T, Ferrone S, Takata M, Uhara H, Okuyama R
  • Journal Title: British J Cancer Volume: 106 Pages: 939-946
  • Peer Reviewed
• [Journal Article] Assessment of BRAF and KIT mutations in Japanese melanoma patients (2012)
  • Author(s): Ashida A, Uhara H, Kiniwa Y, Oguchi M, Murata H, Goto Y, Uchiyama A, Ogawa E, Hayashi K, Koga H, Okuyama R
  • Journal Title: J Dermatol Sci Volume: 66 Issue: 3 Pages: 240-242
  • DOI: 10.1016/j.jdermsci.2012.03.005
  • Peer Reviewed
• [Presentation] Assessment of NRAS mutation in Japanese melanoma patients (2013)
  • Author(s): Ashida A, Uhara H, Okuyama R
  • Organizer: The 72th Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: Yokohama, JPN
• [Presentation] 日本人のメラノーマ患者におけるBRAF、NRAS、KITの遺伝子変異の解析 (2013)
  • Author(s): 芦田敦子、宇原久、奥山隆平
  • Organizer: 日本皮膚悪性腫瘍学会
  • Place of Presentation: 甲府
• [Presentation] Assessment of NRAS mutation in Japanese melanoma patients (2013)
  • Author(s): Ashida A, Uhara H, Okuyama R
  • Organizer: The 72th Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: 横浜
• [Presentation] Assesment of NRAS mutations in Japanese melanoma patients (2013)
  • Author(s): Atsuko Ashida
  • Organizer: 第72回日本癌学会
  • Place of Presentation: 横浜
• [Presentation] 日本人のメラノーマにおけるBRAF, NRAS, KITの遺伝子変異の解析 (2013)
  • Author(s): 芦田敦子
  • Organizer: 第29回皮膚悪性腫瘍学会
  • Place of Presentation: 甲府
• [Presentation] Assessment of BRAF and KIT mutation in Japanese melanoma patients (2012)
  • Author(s): Ashida A, Uhara H, Murata H, Goto Y, Okuyama R
  • Organizer: The 71th Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: Sapporo, JPN
• [Presentation] Assesment of BRAF and KIT mutations in Japanese melanoma patients (2012)
  • Author(s): Atsuko Ashida
  • Organizer: 第71回日本癌学会
  • Place of Presentation: 札幌
• [Book] 皮膚の悪性腫瘍II悪性黒色腫:分子標的薬KITの変異に基づく治療薬,皮膚科臨床アセット17 (2014)
  • Author(s): 芦田敦子、奥山隆平
• [Book] 皮膚科臨床アセット 分子標的薬 KITの変異に基づく治療薬 (2014)
  • Author(s): 芦田敦子、奥山隆平
  • Publisher: 中山書店