癌幹細胞を標的とした糖鎖改変膵癌ワクチン療法の開発と次世代型抗体医薬の創薬

2011 Fiscal Year Annual Research Report

• Project/Area Number: 22591520
• Research Institution: Department of Clinical Research, National Hospital Organization Kure Medical Center
• Principal Investigator: 種村 匡弘 独立行政法人国立病院機構呉医療センター, 外科, 医長 (30379250)
• Co-Investigator(Kenkyū-buntansha): 土岐 祐一郎 大阪大学, 医学研究科, 教授 (20291445) ; 三善 英和 大阪大学, 医学研究科, 教授 (20322183)
• Keywords: 膵癌 / 免疫療法 / 癌幹細胞 / 糖鎖 / 創薬

Research Abstract

【研究目的】糖鎖改変膵がん細胞を用いたワクチン研究を行い、膵癌の癌幹細胞にも有効に抗がん作用を発揮できるかを検討した。
【研究方法】細胞表面にがん抗原・MUC1を発現するヒト膵癌細胞株PANC1にマウスα1,3ガラクトース転移酵素遺伝子をlipofectionにて導入しα-gal MUC1を発現したα-gal PANC1細胞を樹立する。α1,3GTノックアウトマウス(α1,3GT K.O.mice)をブタ腎臓組織片にて免疫しhigh Anti-Gal K.O.miceとしヒトと同じ免疫状態とする。このhigh Anti-Galマウスに1×10^6個のα-gal-PANC1を用いて免疫し(α-gal group)、同様に1×10^6個の野生PANC1を用いて免疫したコントロール群と比し、膵癌の癌幹細胞とされるCD44+CD24+ESA+陽性膵がん細胞に対し抗体を産生しているかを免疫したマウス血清を用いたFACSにて解析した。さらにWestern blotを行い、糖鎖改変ワクチンにてMUC1など明らかな癌抗原に対する抗原だけでなく、膵癌細胞に含まれる未知の癌抗原に対しても抗体を産生しているか解析した。
【結果】磁気ビーズを用いてCD44+CD24+ESA+陽性PANC1細胞を分離し、ワクチン化したKO miceの血清と反応させFACS解析した。α-gal groupの血清ではmean shift intensityで181.9と強い結合を認めたが、control groupでは10.3とほとんど有意な結合性を示さなかった。ウエスタンブロットではcontrol groupの血清では有意なバンドは認めなかったが、α-gal groupでは250KDa以上の高分子に対し抗体産生を認め、MUC1以外の癌抗原にも抗体産生を惹起していることが証明された。
【結語】糖鎖改変膵がん細胞を用いたワクチンでは癌幹細胞に対しても抗体産生を誘導し、さらに未知の癌抗原にも抗体産生を惹起させ、膵癌根治を実現させる免疫療法であることが示唆された。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

これまで、研究計画に沿っておおむね順調に進んでいると考えられる。来年度はin vivoを中心にワクチンの有効性証明に全力を尽くす。

Strategy for Future Research Activity

これまでの実験にて、糖鎖改変膵がん細胞を用いたワクチンはin vitroのデーター上、非常に有効であることが示唆された。来年度は、この抗腫瘍効果を実際の生体内(in vivo)においても有効に作用するかを検証する。

Research Products

• [Journal Article] Prognosis of hepatocellular carcinoma with biliary tumor thrombi following liver surgery (2011)
  • Author(s): Noda T, Nagano H, Tomimaru Y, Murakami M, Wada H, Kobayashi S, Marubashi S, Eguchi H, Takeda Y, Tanemura M, Umeshita K, Kim T, Wakasa K, Doki Y, Mori M
  • Journal Title: Surgery Volume: 49(3) Pages: 371-7
• [Journal Article] Diagnostic value of FDG-PET for lymph node metastasis and outcome of surgery for biliary cancer (2011)
  • Author(s): Kobayashi S, Nagano H, Hoshino H, Wada H, Marubashi S, Eguchi H, Takeda Y, Tanemura M, Kim T, Shimosegawa E, Hatazawa J, Doki Y, Mori M
  • Journal Title: J Surg Oncol Volume: 103(3) Pages: 223-9
  • Peer Reviewed
• [Journal Article] Tyrosine kinase inhibitor PTK/ZK enhances the anti-tumor effects of interferon-α/5-fluorouracil therapy for hepatocellular carcinoma cells (2011)
  • Author(s): Murakami M, Kobayashi S, Marubashi S, Tomimaru Y, Noda T, Wada H, Eguchi H, Takeda Y, Tanemura M, Umeshita K, Doki Y, Mori M, Hiroaki Nagano H
  • Journal Title: Ann Surg Oncol Volume: 18(2) Pages: 589-96
  • Peer Reviewed
• [Journal Article] A thick pancreas is a risk factor for pancreatic fistula after a distal pancreatectomy : Selection of the closure technique according to the thickness (2011)
  • Author(s): Eguchi H, Nagano H, Tanemura M, Takeda Y, Marubashi S, Kobayashi S, Wada H, Umeshita K, Mori M, Doki Y
  • Journal Title: Dig Surg Volume: 28(1) Pages: 50-6
  • Peer Reviewed
• [Journal Article] Clinical Significance of Alpha-Fetoprotein mRNA in Peripheral Blood in Liver Resection for Hepatocellular Carcinoma (2011)
  • Author(s): Marubashi S, Nagano H, Wada H, Kobayashi S, Eguchi H, Takeda Y, Tanemura M, Umeshita K, Doki Y, Mori M
  • Journal Title: Ann Surg Oncol Volume: 18(8) Pages: 2200-9
  • Peer Reviewed
• [Journal Article] IGFBP7 downregulation is associated with tumor progression and clinical outcome in hepatocellular carcinoma (2011)
  • Author(s): Tomimaru Y, Eguchi H, Wada H, Kobayashi S, Marubashi S, Tanemura M, Umeshita K, Kim T, Wakasa K, Doki Y, Mori M, Nagano H
  • Journal Title: Int J Cancer Volume: 15;130(2) Pages: 319-27
  • Peer Reviewed
• [Journal Article] Donor Hepatectomy for Living Donor Liver Transplantation : Learning Steps and Surgical Outcome (2011)
  • Author(s): Marubashi S, Nagano H, Wada H, Kobayashi S, Eguchi H, Takeda Y, Tanemura M, Doki Y, Mori M
  • Journal Title: Dig Dis Sci Volume: 56(8) Pages: 2482-90
  • Peer Reviewed
• [Journal Article] Synergistic antitumor effect of interferon-ss with gemcitabine in interferon-α-non-responsive pancreatic cancer cells (2011)
  • Author(s): Tomimaru Y, Eguchi H, Wada H, Tomokuni A, Kobayashi S, Marubashi S, Takeda Y, Tanemura M, Umeshita K, Mori M, Doki Y, Nagano H
  • Journal Title: Int J Oncol Volume: 38(5) Pages: 1237-43
  • Peer Reviewed
• [Journal Article] Mural nodule in branch duct-type intraductal papillary mucinous neoplasms of the pancreas is a marker of malignant transformation and indication for surgery (2011)
  • Author(s): Akita H, Takeda Y, Hoshino H, Wada H, Kobayashi S, Marubashi S, Eguchi H, Tanemura M, Mori M, Doki Y, Nagano H
  • Journal Title: Am J Surg Volume: 202(2) Pages: 214-9
  • Peer Reviewed
• [Journal Article] Combination use of anti-CD133 antibody and SSA lectin can effectively enrich cells with high tumorigenicity (2011)
  • Author(s): Moriwaki K, Okudo K, Haraguchi N, Takeishi S, Sawaki H, Narimatsu H, Tanemura M, Ishii H, Mori M, Miyoshi E
  • Journal Title: Cancer Sci Volume: 102(6) Pages: 1164-70
  • Peer Reviewed
• [Journal Article] Ep-CAM is a significant prognostic factor in pancreatic cancer patients by suppressing cell activity (2011)
  • Author(s): Akita H, Nagano H, Takeda Y, Eguchi H, Wada H, Kobayashi S, Marubashi S, Tanemura M, Takahashi H, Ohigashi H, Tomita Y, Ishikawa O, Mori M, Doki Y
  • Journal Title: Oncogene Volume: 4;30(31) Pages: 3468-76
  • Peer Reviewed
• [Journal Article] Successful pregnancy after simultaneous pancreas-kidney transplantation from a brain-dead donor : the first case report in Japan (2011)
  • Author(s): Koyama S, Tomimatsu T, Kanagawa T, Daimon E, Kimura T, Kuroda A, Tanemura M, Doki Y, Ito T
  • Journal Title: J Obstetrics Gynaecol Res Volume: 37(11) Pages: 1711-6
  • Peer Reviewed
• [Journal Article] Long-term outcome of combined subcutaneous interferon-α and intra-arterial 5-fluorouracil treatment for advanced hepatocellular carcinoma with major portal vein thrombosis (2011)
  • Author(s): Nagano H, Wada H, Kobayashi S, Marubashi S, Eguchi H, Tanemura M, Tomimaru Y, Osuga K, Umeshita K, Doki Y, Mori M
  • Journal Title: Oncology Volume: 80(1-2) Pages: 63-9
  • Peer Reviewed
• [Journal Article] Circulating microRNA-21 as a novel biomarker for hepatocellular carcinoma (2011)
  • Author(s): Tomimaru Y, Eguchi H, Nagano H, Wada H, Kobayashi S, Marubashi S, Tanemura M, Tomokuni A, Takemasa I, Umeshita K, Kanto T, Doki Y, Mori M
  • Journal Title: J Hepatol Volume: 56(1) Pages: 167-75
  • Peer Reviewed
• [Journal Article] Significance of Alanine Aminopeptidase N (APN) in Bile in the Diagnosis of Acute Cellular Rejection After Liver Transplantation (2011)
  • Author(s): Kim C, Aono S, Marubashi S, Wada H, Kobayashi S, Eguchi H, Takeda Y, Tanemura M, Okumura N, Takao T, Doki Y, Mori M, Nagano H
  • Journal Title: J Surg Res Volume: Mar 25(Epub ahead of print)
  • Peer Reviewed
• [Journal Article] Treatment of borderline cases for curative resection of biliary tract cancer (2011)
  • Author(s): Kobayashi S, Nagano H, Marubashi S, Wada H, Eguchi H, Takeda Y, Tanemura M, Umeshita K, Doki Y, Mori M
  • Journal Title: J Surg Oncol Volume: 104(5) Pages: 499-503
  • Peer Reviewed
• [Journal Article] Reprogramming of mouse and human cells to pluripotency using mature microRNAs (2011)
  • Author(s): Miyoshi N, Ishii H, Tanemura M, Sekimoto M, Doki Y, Mori M
  • Journal Title: Cell Stem Cell Volume: 3;8(6) Pages: 633-8
  • Peer Reviewed
• [Journal Article] Experience with the use of fibrin sealant plus polyglycolic acid felt at the cut surface of the liver in laparoscopic hepatectomy (2011)
  • Author(s): Kobayashi S, Nagano H, Marubashi S, Wada H, Takeda Y, Eguchi H, Tanemura M, Umeshita K, Doki Y, Mori M
  • Journal Title: Surg Endosc Volume: 25(11) Pages: 3590-6
  • Peer Reviewed
• [Journal Article] Successful adult ABO incompatible living donor liver transplantation : experience with double infusion through the hepatic artery and portal vein (2011)
  • Author(s): Kobayashi S, Nagano H, Marubashi S, Wada H, Takeda Y, Eguchi H, Tanemura M, Umeshita K, Doki Y, Mori M
  • Journal Title: Hepatogastroenterology Volume: 58(106) Pages: 503-7
  • Peer Reviewed
• [Journal Article] Role of the Hypoxia-Related Gene, JMJD1A, in Hepatocellular Carcinoma : Clinical Impact on Recurrence after Hepatic Resection (2011)
  • Author(s): Yamada D, Kobayashi S, Yamamoto H, Tomimaru Y, Noda T, Uemura M, Wada H, Marubashi S, Eguchi H, Tanemura M, Doki Y, Mori M, Nagano H
  • Journal Title: Ann Surg Oncol Volume: May 24(Epub ahead of print)
  • Peer Reviewed
• [Presentation] Vaccination with pancreatic cancer cells expressing α-gal epitopes effectively induced immuneresponses against not only differentiated cancer cells but also cancer stem cells (2011)
  • Author(s): Masahiro Tanemura
  • Organizer: AACR総会
  • Place of Presentation: フロリダ
  • Year and Date: 20110400
• [Presentation] Preliminary experiment of α-gal epitopes immune-based therapy against pancreatic cancer, by tumor cell lysates (2011)
  • Author(s): Masahiro Tanemura
  • Organizer: AACR総会
  • Place of Presentation: フロリダ
  • Year and Date: 20110400
• [Presentation] 遠隔成績からみた1型糖尿病に対する脳死膵臓移植の現状と再生医療を応用した次世代型糖尿病根治療法の開発 (2011)
  • Author(s): 種村匡弘
  • Organizer: 第14回阪神肝胆膵疾患研究会
  • Place of Presentation: 甲子園ノボテル
  • Year and Date: 2011-12-02
• [Presentation] 遠隔成績からみた1型糖尿病に対する脳死膵臓移植の現状と明日の糖尿病再生医療を構築する基礎研究 (2011)
  • Author(s): 種村匡弘
  • Organizer: 第253回呉消化器病研究会
  • Place of Presentation: 呉阪急ホテル
  • Year and Date: 2011-10-15
• [Presentation] ブタ型糖鎖(α-galエピトープ)への改変による新しい膵がんワクチン療法の開発と臨床応用 (2011)
  • Author(s): 種村匡弘
  • Organizer: 関西グライコサイエンス
  • Place of Presentation: 大阪大学
  • Year and Date: 2011-05-14