Development of novel gene-therapy using IL-12 and DNA nanoassociate bearing hyaluronan-glycocalyx for recurrent hepatocellular carcinoma after liver transplantation

• Project/Area Number: 20591528
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General surgery
• Research Institution: Kyushu University
• Principal Investigator: SOEJIMA Yuji Kyushu University, 統括診療部, 消化器外科医長 (30325526)
• Co-Investigator(Kenkyū-buntansha): MAEHARA Yoshihiko 九州大学, 教授 (80165662) ; TAKETOMI Akinobu 九州大学, 助教 (70363364) ; 吉住 朋晴 九州大学, 大学病院, 特別教員 (80363373)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2010: ¥130,000 (Direct Cost: ¥100,000、Indirect Cost: ¥30,000); Fiscal Year 2009: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2008: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
• Keywords: 移植外科学 / 肝細胞癌 / 遺伝子治療 / インターロイキン12 / ヒアルロン酸くし型共重合体

Research Abstract

In this project, we investigated the effectiveness of gene therapy using IL-12 and DNA nanoassociate bearing hyaluronan-glycocalyx argeting the sinusoidal endothelial cells for the treatment of recurrent hepatocellular carcinoma after liver transplantation. First of all, mIL-12 plasmid DNA was purified. To develop a system for targeting foreign DNA to SECs, comb-type polycations having HA side chains were prepared by coupling HA to poly(L-lysine) (PLL). The HA-grafted-PLL copolymer (PLL-g-HA) thus formed was mixed with DNA in 154 mM NaCl to form soluble nanoassociates bearing hydrated hyaluronate shells. Agarose gel retardation assays revealed selective interaction of the PLL backbone with DNA despite the presence of polyanionic HA side chains.PLL-g-HA. Polymerization of PLL-g-HA with mIL-12 plasmid DNA was then confirmed. However, stable production and supply of PLL-g-HA/pCAGGS-mIL-12 was difficult due to the lack of PLL-g-HA. C3H mice (H-2(k)) were s.c. implanted with 2.5 x 10(6) MH134 cells (H-2(k)) and we treated the established HCC with electroporation-mediated gene therapy using mIL-12 plasmid DNA. Intratumoral gene transfer of mIL-12 elevated intratumoral mIL-12, IFN-gamma, and IFN-gamma-inducible protein-10, significantly inhibited the growth of HCC, compared with HCC-transferred control pCAGGS plasmid.

Research Products

• [Journal Article] "All-in-one" ex vivo self-reconstruction technique using an autologous inferior vena cava for a right lobe liver graft with multiple and complex venous orifices. (2010)
  • Author(s): Soejima Y., Takeishi K., Ikegami T., Uchiyama H., Taketomi A., Maehara Y.
  • Journal Title: Liver Transpl. 16 Pages: 909-913
• [Journal Article] "All-in-one" ex vivo self-reconstruction technique using an autologous inferior vena cava for a right lobe liver graft with multiple and complex venous orifices. (2010)
  • Author(s): Soejima Y., Takeishi K., Ikegami T., Uchiyama H., Taketomi A., Maehara Y.
  • Journal Title: Liver Transpl. Volume: 16 Pages: 909-913
  • Peer Reviewed
• [Journal Article] Clinicopathological prognostic factors after hepatectomy for patients with mass-forming type intrahepatic cholangiocarcinoma : relevance of the lymphatic invasion index. (2010)
  • Author(s): Shirabe K., Mano Y., Taketomi A., Soejima Y., Uchiyama H., Aishima S., Kayashima H., Ninomiya M., Maehara Y.
  • Journal Title: Ann Surg Oncol. Volume: 17 Pages: 1816-1822
  • Peer Reviewed
• [Journal Article] Improved results of a surgical resection for the recurrence of hepatocellular carcinoma after living donor liver transplantation. (2010)
  • Author(s): Taketomi A., Fukuhara T., Morita K., Kayashima H., Ninomiya M., Yamashita Y., Ikegami T., Uchiyama H., Yoshizumi T., Soejima Y., Shirabe K., Maehara Y.
  • Journal Title: Ann Surg Oncol. Volume: 17 Pages: 2283-2289
  • Peer Reviewed
• [Journal Article] The use of autologous vena cava patch graft for reconstructing multiple outflow orifices in right hemiliver living donor graft (2009)
  • Author(s): Ikegami T, Soejima Y, Nagata S, Taketomi A, Maehara Y.
  • Journal Title: Transplant Int 22 Pages: 853-5
  • Peer Reviewed
• [Journal Article] Hypersplenism after living donor liver transplantation (2009)
  • Author(s): Ikegami T, Soejima Y, Taketomi A, Kawanaka H, Yoshizumi T, Shimada M, Maehara Y.
  • Journal Title: Hepatogastroenterology 56 Pages: 778-82
  • Peer Reviewed
• [Journal Article] Recurrent familial hypobetalipoproteinemia-induced nonalcoholic fatty liver disease after living donor liver transplantation (2009)
  • Author(s): Harada N, Soejima Y, Taketomi A, Yoshizumi T, Uchiyama H, Ikegami T, Saibara T, Nishizaki T, Maehara Y.
  • Journal Title: Liver Transpl 15 Pages: 806-9
  • Peer Reviewed
• [Journal Article] A simple hilar dissection technique preserving maximum blood supply to the bile duct in living donor liver transplantation. (2008)
  • Author(s): Soejima Y., Fukuhara T., Morita K., Yoshizumi T., Ikegami T.,Yamashita Y., Sugimachi K., Taketomi A., Maehara Y
  • Journal Title: Transplantation. 86(9) Pages: 1468-1469
• [Journal Article] (2008)
  • Author(s): Soejima Y., Meguro M., Taketomi A., Ikegami T., Yamashita Y., Harada N., Ito S., Uchiyama H., Yoshizumi T., Maehara Y.
  • Journal Title: Transpl Int. 21(4) Pages: 384-389
• [Journal Article] Living donor liver transplantation using dual grafts from two donors: a feasible option to overcome small-for-size problems? (2008)
  • Author(s): Soejima Y., Taketomi A., Ikegami T., Yoshizumi T., Uchiyama H., Yamashita Y., Meguro M., Harada N., Shimada M., Maehara Y.
  • Journal Title: Am J Transplant. 8(4) Pages: 887-892
• [Journal Article] One-step venous reconstruction for a right lobe graft with multiple vein orifices in living donor liver transplantation. (2008)
  • Author(s): Soejima Y., Ueda N., Fukuhara T., Yoshizumi T., Ikegami T., Yamashita Y., Sugimachi K., Taketomi A., Maehara Y.
  • Journal Title: Liver Transpl. 14(5) Pages: 706-708
• [Journal Article] Sequential pancreatoduodenectomy after living donor liver transplantation for cholangiocarcinoma. (2008)
  • Author(s): Soejima Y., Ueda S., Sanefuji K., Kayashima H., Yoshizumi T., Ikegami T., Yamashita Y., Sugimachi K., Iguchi T., Taketomi A., Maehara Y.
  • Journal Title: Am J Transplant. 8(10) Pages: 2158-2162
• [Presentation] Extended indication for living donor liver transplantation in patients with hepatocellular carcinoma : Kyushu University experience using a newly developed risk score. (2010)
  • Author(s): Soejima Y., Shirabe K., Taketomi A., Yoshizumi T., Uchiyama H., Ikegami T., Maehara Y.
  • Organizer: 第110回日本外科学会定期学術集会総会
  • Place of Presentation: 名古屋
• [Presentation] All-in-one ex vivo self-reconstruction technique using an autologous inferior vena cava for a right lobe graft with multiple and complex venous orifices. (2010)
  • Author(s): Soejima Y., Shirabe K, Taketomi A., Yoshizumi T., Uchiyama H., Ikegami T., Ninomiya M., Maehara Y.
  • Organizer: ILTS第16回国際肝臓移植学会
  • Place of Presentation: 香港
• [Presentation] 生体肝移植における摘出肝門脈及び内頚静脈グラフトを用いた血行再建手技とその応用 (2009)
  • Author(s): 副島雄二, 武冨紹信, 内山秀昭, 池上徹, 二宮瑞樹, 萱島寛人, 森田和豊, 戸島剛男, 武石一樹, 本村貴志, 前原喜彦
  • Organizer: 第21回日本肝胆膵外科学会
  • Place of Presentation: 名古屋
• [Presentation] Salvage living donor liver transprantation for recurrent hepatocellular carcinoma. (2008)
  • Author(s): 副島雄二、武冨紹信、吉住朋晴、山下洋市、杉町圭史、前原喜彦
  • Organizer: 第108回日本外科学会国際シンポジウム
  • Place of Presentation: 長崎
• [Book] 腹腔鏡下肝切除術と知っておきたい高難易度肝切除術 (2010)
  • Author(s): 副島雄二、調憲、矢永勝彦、前原喜彦
  • Total Pages: 14
  • Publisher: メディカルビュー社
• [Book] 肝移植四半世紀の歩み (2009)
  • Author(s): 副島雄二, 前原喜彦
  • Total Pages: 78
  • Publisher: 九州大学における肝移植のあゆみ