Generation and characterization of human heme oxygenase-1 transgenic pigs.
Xenotransplantation using transgenic pigs as an organ source is a promising strategy to overcome shortage of human organ for transplantation. Various genetic modifications have been tried to ameliorate xenograft rejection. In the present study we assessed effect of transgenic expression of human heme oxygenase-1 (hHO-1), an inducible protein capable of cytoprotection by scavenging reactive oxygen species and preventing apoptosis caused by cellular stress during inflammatory processes, in neonatal porcine islet-like cluster cells (NPCCs). Transduction of NPCCs with adenovirus containing hHO-1 gene significantly reduced apoptosis compared with the GFP-expressing adenovirus control after treatment with either hydrogen peroxide or hTNF-α and cycloheximide. These protective effects were diminished by co-treatment of hHO-1 antagonist, Zinc protoporphyrin IX. We also generated transgenic pigs expressing hHO-1 and analyzed expression and function of the transgene. Human HO-1 was expressed in most tissues, including the heart, kidney, lung, pancreas, spleen and skin, however, expression levels and patterns of the hHO-1 gene are not consistent in each organ. We isolate fibroblast from transgenic pigs to analyze protective effect of the hHO-1. As expected, fibroblasts derived from the hHO-1 transgenic pigs were significantly resistant to both hydrogen peroxide damage and hTNF-α and cycloheximide-mediated apoptosis when compared with wild-type fibroblasts. Furthermore, induction of RANTES in response to hTNF-α or LPS was significantly decreased in fibroblasts obtained from the hHO-1 transgenic pigs. These findings suggest that transgenic expression of hHO-1 can protect xenografts when exposed to oxidative stresses, especially from ischemia/reperfusion injury, and/or acute rejection mediated by cytokines. Accordingly, hHO-1 could be an important candidate molecule in a multi-transgenic pig strategy for xenotransplantation.
Transplantation Research Institute, College of Medicine, Seoul National University, Seoul, Korea.
Hye-Jung
HJ
Yeom
Ok Jae
OJ
Koo
Jaeseok
J
Yang
Bumrae
B
Cho
Jong-Ik
JI
Hwang
Sol Ji
SJ
Park
Sunghoon
S
Hurh
Hwajung
H
Kim
Eun Mi
EM
Lee
Han
H
Ro
Jung Taek
JT
Kang
Su Jin
SJ
Kim
Jae-Kyung
JK
Won
Philip J
PJ
O'Connell
Hyunil
H
Kim
Charles D
CD
Surh
Byeong-Chun
BC
Lee
Curie
C
Ahn
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D017209:Apoptosis; D002470:Cell Survival; D002478:Cells, Cultured; D016207:Cytokines / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000201:enzymology / Q000276:immunology / Q000502:physiology; D005818:Genetic Engineering; D051547:Heme Oxygenase-1 / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D006801:Humans; D007515:Islets of Langerhans / Q000201:enzymology / Q000502:physiology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D009928:Organ Specificity; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D034421:Sus scrofa / Q000235:genetics*; D014183:Transplantation, Heterologous; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology / Q000502:physiology
D016207:Cytokines; D008070:Lipopolysaccharides; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D051547:Heme Oxygenase-1