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2000 Fiscal Year Final Research Report Summary

In vivo liver-directed gene transfer using in-situ perfusion of the porcine liver

Research Project

Project/Area Number 09307027
Research Category

Grant-in-Aid for Scientific Research (A).

Allocation TypeSingle-year Grants
Section一般
Research Field Digestive surgery
Research InstitutionUniversity of Ryukyus

Principal Investigator

SHIRAISHI Masayuki  hospital attached to the college, University of Ryukyus, lecturer, 医学部・附属病院, 講師 (00264482)

Co-Investigator(Kenkyū-buntansha) HIROYASU Shunngo  the college of medicine, University of Ryukyus assistant, 医学部, 助手 (90295337)
MUTO Yoshihiro  the college of medicine, University of Ryukyus professor, 医学部, 教授 (60157724)
Project Period (FY) 1997 – 2000
Keywordsadenovirus vector / gene transfer / pig / rat / von Willebrand factor / HVJ-liposome
Research Abstract

【Aims】 The aim of this study was to establish the effective gene transfer to a porcine liver in-vivo, and to utilize this model for gene therapy for metabolic disorder (a deficiency or abnormality of certain proteins or enzymes). In this setting, the transfected liver was expected to produce a target protein after successful gene transfer. To accomplish these aims, the von Willebrand factor (vWF) gene was transferred into the livers in-vivo, in pigs with and without von Willebrand disease (vWD), and the plasma vWF levels were thereafter assessed. Since vWF is produced and secreted from the endothelial cells, the vWF gene should thus be transferred into the sinusoidal endothelial cells in the pig liver.
【Results】 Preliminary studies have demonstrated that both the marker LacZ gene and human vWF were effectively transferred into the porcine endothelial cells in-vitro (≧ 90%), either with adenovirus vector (AdV) or HVJ-liposome vector (HVJ). On the other hand, neither the systemic injectio … More n nor the intraportal injection could establish any gene transfer (LacZ, vWF) in the in-vivo pig liver, using the same vectors (AdV or HVJ). We next tested in-situ perfusion of the liver (ISP) as a method for in-vivo gene transfer, in which the liver was isolated from systemic circulation under a pump driven bypass of the blood flow from the portal vein and inferior vena cava, and the liver was perfused with a high titer AdV or HVJ under bloodless and cold ischemic conditions. As a result, the marker LacZ expression was confirmed (X-gal staining & RT-PCR) in both the hepatocytes and sinusoidal endothelial cells in the pig liver. The mRNA expression of transferred vWF (HVJ) was also confirmed by RT-PCR only after ISP.The plasma vWF levels, however, did not significantly increase after gene transfer.
【Conclusion】 The in-vivo gene transfer of vWF was thought to be possible by an ISP of the liver and HVJ-liposome vector. A significant increase of vWF, however, could not be confirmed, thus suggesting that further investigations are thus needed to establish the clinically relevant levels of gene transfer. Less

  • Research Products

    (47 results)

All Other

All Publications (47 results)

  • [Publications] Nozato E: "In-situ perfusion of the liver under portosystemic shunt in rats : a stable model of cold ischemia for orthotopic liver transplantation"Transplantation Proc. 30・7. 3718-3720 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "In-situ perfusion of the liver as a stable model of cold ischemia in orthotopic liver transplantation"Ryukyu Med J. 18・3. 73-77 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Early Detection of the Fas-FasL System in Hepatic Graft-Versus Host Disease After Rat Small-Bowel Transplantation"Transplantation Proc. 31・1-2. 567-568 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Stable Gene Expression with VSV-G Pseudotyped-Retrovirus Vector in the Rat Liver"Journal of Surgical Research. 84・2. 168-173 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiroyasu S: "Analysis of the Fas System and Bcl-2 in Rat Liver Allograft Rejection"Journal of Surgical Research. 84・2. 204-211 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 白石祐之: "ブタ肝臓異種灌流モデルにおける複数ヒト補体抑制蛋白遺伝子導入"外科治療. 81・3. 366-367 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer using in-situ perfusion of the liver graft"Transplant International. 10・3. 202-206 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Graft-directed gene transfection in a rat model of liver transplantation"Ryukyu Med J. 17・1. 21-24 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Successful Gene Transfer to the Porcine Liver in Vivo with an Adenoviral Vector"Journal of Surgical Research. 76・2. 105-110 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Obuchi Y: "Highly Effective Adenoviral Mediated Gene Transfer to Porcine Endothelia Cells In Vitro"Transplantation Proc. 30・7. 2917-2918 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagahama M: "Adenoviral Mediated Gene Transfer to the Porcine Liver in Vivo Using In Situ Perfusion of the Liver"Transplantation Proc. 30・7. 2914-2916 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Taira K: "A New, Stable Model of Left Lobectomy for Living-Related Liver Transplantation in the Pig"Transplantation Proc. 30・7. 3207-3208 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Early Detection of Apoptosis and Fas Ligand Expression in Hepatic Graft-versus-Host Disease after Rat Small Bowel Transplantation"Journal of Surgical Research. 86・1. 136-144 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiroyasu S: "Analysis of FAS System in Pulmonary Injury of Graft-Versus-Host Disease after Rat Intestinal Transplantation"Transplantation. 68・7. 933-938 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okuhama Y: "Protective effects of ulinastatin against ischemia-reperfusion injury"Journal of Surgical Research. 82・1. 34-42 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomori H: "Protective effects lidocaine in hepatic ischemia/reperfusion injury in-vivo and ex-vivo"Ryukyu Med J. 19・1. 11-16 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomori H: "In situ perfusion of the liver Enhances the Efficiency of retrovirusmediated gene transfer to hepatocytes"Eur Surg Res. 32・1. 18-22 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Higa T: "Limitations of Exogenous L-Arginine in Exerting a Cytoprotective Effect on Hepatic Ischemia/Reperfusion Injury"Surg Today. 30・4. 352-359 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mamadi T: "Effect of Bcl-2 expression on morphological alteration of hepatic mitochondria in liver regeneration after a partial hepatectomy"Ryukyu Med J. 19・4. 203-209 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagahama M: "Adenovirus-mediated gene transfer of double human complement regulating proteins ・DAF, CD59) in xenogeneic rat kidney perfusion"Transplant Proc. 32・5. 909-910 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer of the triple human complement regulating proteins ・DAF, CD59, MCP) in xenogeneic pig liver perfusion"Transplant Proc. 32・5. 1116-1117 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer of Bcl-2 in graft-versus-host disease after rat intestinal transplantation"Transplant Proc. 32・6. 1263-1264 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer of DAF and CD59 in xenogeneic pig liver perfusion"Transplant Proc. 32・7. 2374-2375 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagahama M: "Adenovirus-mediated triple gene transfer of human complement regulating proteins to the porcine endothelial cell"Transplant Proc. 32・7. 2511-2513 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer using in-situ perfusion of the liver graft"Transplant International. 10 3. 202-206 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Graft-directed gene transfection in a rat model of liver transplantation"Ryukyu Med J. 17 1. 21-24 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Successful Gene Transfer to the Porcine Liver in Vivo with an Adenoviral Vector"Journal of Surgical Research. 76 2. 105-110 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Obuchi Y: "Highly Effective Adenoviral Mediated Gene Transfer to Porcine Endothelial Cells In Vitro"Transplantation Proc. 30 7. 2917-2918 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagahama M: "Adenoviral Mediated Gene Transfer to the Porcine Liver in Vivo Using in Situ Perfusion of the Liver"Transplantation Proc. 30 7. 2914-2916 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Taira K: "A New, Stable Model of Left Lobectomy for Living-Related Liver Transplantation in the Pig"Transplantation Proc. 30 7. 3207-3208 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nozato E: "In-situ perfusion of the liver under portosystemic shunt in rats : a stable model of cold ischemia for orthotopic liver transplantation"Transplantation Proc. 30 7. 3718-3720 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "In-situ perfusion of the liver as a stable model of cold ischemia in orthotopic liver transplantation"Ryukyu Med J. 18 3. 73-77 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Early Detection of the Fas-FasL System in Hepatic Graft-Versus Host Disease After Rat Small-Bowel Transplantation"Transplantation Proc. 31 1-2. 567-568 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Stable Gene Expression with VSV-G Pseudotyped-Retrovirus Vector in the Rat Liver"Journal of Surgical Research. 84 2. 168-173 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiroyasu S: "Analysis of the Fas System and Bcl-2 in Rat Liver Allograft Rejection"Journal of Surgical Research. 84 2. 204-211 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Early Detection of Apoptosis and Fas Ligand Expression in Hepatic Graft-versus-Host Disease after Rat Small Bowel Transplantation"Journal of Surgical Research. 86 1. 136-144 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiroyasu S: "Analysis of FAS System in Pulmonary Injury of Graft-Versus-Host Disease after Rat Intestinal Transplantation"Transplantation. 68 7. 933-938 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Okuhama Y: "Protective effects of ulinastatin against ischemia-reperfusion injury"Journal of Surgical Research. 82 1. 34-42 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomori H: "Protective effects lidocaine in hepatic ischemia/reperfusion injury-in-vivo and ex-vivo"Ryukyu Med J. 19 1. 11-16 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomori H: "In situ perfusion of the liver Enhances the Efficiency of retrovirus-mediated gene transfer to hepatocytes"Eur Surg Res. 32 1. 18-22 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Higa T: "Limitations of Exogenous L-Arginine in Exerting a Cytoprotective Effect on Hepatic Ischemia/Reperfusion Injury"Surg Today. 30 4. 352-359 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mamadi T: "Effect of Bcl-2 expression on morphological alteration of hepatic mitochondria in liver regeneration after a partial Hepatectomy"Ryukyu Med J. 19 4. 203-209 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagahama M: "Adenovirus-mediated gene transfer of double human complement regulating proteins DAF, CD59) in xenogeneic rat kidney perfusion"Transplant Proc. 32 5. 909-910 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer of the triple human complement regulating proteins DAF, CD59, MCP) in xenogeneic pig liver perfusion"Transplant Proc. 32 5. 1116-1117 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer of Bcl-2 in graft-versus-host disease after rat intestinal transplantation"Transplant Proc. 32 6. 1263-1264 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi M: "Adenovirus-mediated gene transfer of DAF and CD59 in xenogeneic pig liver perfusion"Transplant Proc. 32 7. 2374-2375 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagahama M: "Adenovirus-mediated triple gene transfer of human complement regulating proteins to the porcine endothelial cell"Transplant Proc. 32 7. 2511-2513 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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