婦人科癌転移の特異的糖鎖構造解析と認識糖鎖抗原をターゲットする新規治療抗体の開発

2015 Fiscal Year Annual Research Report

• Project/Area Number: 24592530
• Research Institution: Keio University
• Principal Investigator: 鈴木 淳 慶應義塾大学, 医学部, 助教 (00255514)
• Co-Investigator(Kenkyū-buntansha): 鈴木 雅美 慶應義塾大学, 医学部, 研究員 (10276321) ; 冨永 英一郎 慶應義塾大学, 医学部, 講師 (80276328)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Keywords: 糖鎖抗原 / 抗体

Outline of Annual Research Achievements

我々が作製したヒト抗体は婦人科癌、中でも子宮体癌に特異的に抗腫瘍効果を示すことを解明した。子宮体癌のリンパ節転移能で差を認める低転移能のHec 8細胞および高転移能のHec 9細胞を用いたin vitro proliferation assayではHMMC-1は両細胞に対して増殖抑制効果を示したが、高い転移能を示すHec 9細胞に対しては、より強い増殖抑制効果を示した。細胞間での細胞周期の影響を除外するため、cell cycle analysisを用いて、cell cycleのマーカーとなるp16, p21, p27の発現には差がないことが判明し、増殖抑制効果は増殖能の差に影響されないことを示した。
さらにHMMC-1抗体の認識糖鎖抗原を解明した。Hec 9細胞をSDS-gel電気泳動し、HMMC-1反応タンパクをさらにUEA-1カラムクロマトグラフィーにて精製した。トリプシン処理後にAXIMA-CFR plus MALDI-TOF質量分析し、MASCOT解析にてそのタンパク同定を行った。その結果、HMMC-1抗体の反応タンパクはCD166およびCD49であることが判明した。これらタンパク上の認識抗原構造がFucα1→2Galβ1→4GlcNAcβ1→3Galβ1→3GalNAc→Ser/Thrであることを解明した。この糖鎖抗原を発現する糖転移酵素のα1,2FucT1, Core1β1,3GlcNAcT, β1,3GalT5, およびβ1,4GalT1の発現解析をHec 8およびHec 9細胞で行い、より転移能が高いHec 9細胞にはextended core 1構造を決定するCore1β1,3GlcNAcT酵素の発現上昇を認めた。局所発現においてもconfocal免疫組織化学染色にてCD166およびHMMC-1抗体がHec 9細胞において同部位を認識していることを解明した。結論として、HMMC-1抗体は子宮体癌細胞の増殖抑制効果をin vitroで示し、その認識糖鎖抗原がFucα1→2Galβ1→4GlcNAcβ1→3Galβ1→3GalNAc→Ser/Thrであり、HMMC-1抗体の認識タンパクはCD166であることがわかり、子宮体癌の中でもCD166発現上昇を示す腫瘍にはHMMC－1抗体は分子標的治療薬としての今後の臨床的役割を検討する。

Research Products

• [Journal Article] Effectiveness of third-line chemotherapy in recurrent ovarian cancer patients (2015)
  • Author(s): Yoshihama T, Chiyoda T, Kataoka F, Nomura H, Iguchi Y, Hashimoto S, Yamagami W, Tominaga E, Susumu N, Tsuda H, Aoki D.
  • Journal Title: Eur J Gynaecol Oncol Volume: 36 (4) Pages: 424-427
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Risk-reducing surgery in hereditary gynecological cancer: Clinical applications in Lynch syndrome and hereditary breast and ovarian cancer. (2015)
  • Author(s): Adachi M, Banno K, Yanokura M, Iida M, Nakamura K, Nogami Y, Umene K, Masuda K, Kisu I, Ueki A, Hirasawa A, Tominaga E, Aoki D
  • Journal Title: Mol Clin Oncol. Volume: 3(2) Pages: 267-273
  • DOI: 10.3892/mco.2014.460
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Drug repositioning for gynecologic tumors: a new therapeutic strategy for cancer. (2015)
  • Author(s): Banno K, Iida M, Yanokura M, Irie H, Masuda K, Kobayashi Y, Tominaga E, Aoki D.
  • Journal Title: ScientificWorldJournal Volume: 2015 Pages: 1-10
  • DOI: 10.1155/2015/341362
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Clinicopathologic analysis with immunohistochemistry for DNA mismatch repair protein expression in synchronous primary endometrial and ovarian cancers. (2015)
  • Author(s): Kobayashi Y, Nakamura K, Nomura H, Banno K, Irie H, Adachi M, Iida M, Umene K, Nogami Y, Masuda K, Kisu I, Ueki A, Yamagami W, Kataoka F, Hirasawa A, Tominaga E, Susumu N, Aoki D.
  • Journal Title: Int J Gynecol Cancer Volume: 25(3) Pages: 440-6
  • DOI: 10.1097/IGC.0000000000000377
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Aurora kinase A has a significant role as a therapeutic target and clinical biomarker in endometrial cancer (2015)
  • Author(s): Umene K, Yanokura M, Banno K, Irie H, Adachi M, Iida M, Nakamura K, Nogami Y, Masuda K, Kobayashi Y, Tominaga E, Aoki D
  • Journal Title: Int J Oncol. Volume: 46(4) Pages: 1498-506
  • DOI: 10.3892/ijo.2015.2842
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Osteoporosis is less frequent in endometrial cancer survivors with hypertriglyceridemia. (2015)
  • Author(s): Hirasawa A, Makita K, Akahane T, Yamagami W, Makabe T, Yokota M, Horiba Y, Ogawa M, Yanamoto S, Deshimaru R, Tominaga E, Banno K, Susumu N, Aoki D
  • Journal Title: Jpn J Clin Oncol Volume: 45(1) Pages: 127-31
  • DOI: 10.1093/jjco/hyu164
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Investigation of double cancer arising in young patients with endometrial cancer who receive fertility-preserving hormonal therapy using medroyprogesterone acetate. (2015)
  • Author(s): Nobuyuki Susumu, Wataru Yamagami, Hiroyuki Nomura, Fumio Kataoka, Yoshikazu Ichikawa, Aya Takigawa, Atsushi Suzuki, Eiichiro Tominaga, Daisuke Aoki.
  • Organizer: 19th International Meeting of the European Society of Gynecological Oncology.
  • Place of Presentation: Nice, France
  • Year and Date: 2015-10-15 – 2015-10-18
  • Int'l Joint Research
• [Presentation] Usefulness of intra-operative touch imprinting cytology of sentinel lymph nodes in endometrial cancer for improving intraoperative diagnostic accuracy (2015)
  • Author(s): Nobuyuki Susumu, Wataru Yamagami, Hiroyuki Nomura, Fumio Kataoka, Yoshikazu Ichikawa, Aya Takigawa, Atsushi Suzuki, Eiichiro Tominaga, Daisuke Aoki
  • Organizer: 39th European Congress of Cytology
  • Place of Presentation: Milan, Italy
  • Year and Date: 2015-09-20 – 2015-09-23
  • Int'l Joint Research
• [Presentation] A highly allosteric AKT inhibitor shows potent antiproliferative activity in combination with carboplatin and placlitaxel on malignant ascites cells from patients with ovarian cancer. (2015)
  • Author(s): Eiichiro Tominaga, Nobuyuki Susumu, Wataru Yamagami, Hiroyuki Nomura, Fumio Kataoka, Atsushi Suzuki, Daisuke Aoki
  • Organizer: 39th European Congress of Cytology
  • Place of Presentation: Milan, Italy
  • Year and Date: 2015-09-20 – 2015-09-23
  • Int'l Joint Research