Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Identification of Endothelial Specific Region in the Intracellular Adhesion Molecule-2 (ICAM2) Promoter of Miniature Pig

  • Jang, Hoon (Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Jang, Won-Gu (Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Dong Un (Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Eun-Jung (Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Hwang, Sung Soo (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Oh, Keon Bong (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Jeong-Woong (Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • Received : 2012.09.10
  • Accepted : 2012.09.18
  • Published : 2012.09.30
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Abstract

The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

Keywords

References

  1. Yang YG, Sykes, M (2007): Xenotransplantation: current status and a perspective on the future. Nat Rev Immunol 7:519-531. https://doi.org/10.1038/nri2099
  2. Cozzi E, Bhatti F, Schmoeckel M, Chavez G, Smith KG, Zaidi A, Bradley JR, Thiru S, Goddard M, Vial C, Ostlie D, Wallwork J, White DJ, Friend PJ (2000): Long-term survival of nonhuman primates receiving life-supporting transgenic porcine kidney xenografts. Transplantation 70:15-21.
  3. Pierson RN, 3rd (2009): Current status of xenotransplantation. JAMA 301:967-969. https://doi.org/10.1001/jama.2009.237
  4. Good AH, Cooper DK, Malcolm AJ, Ippolito RM, Koren E, Neethling FA, Ye Y, Zuhdi N, Lamontagne LR (1992): Identification of carbohydrate structures that bind human antiporcine antibodies: implications for discordant xenografting in humans. Transplant Proc 24: 559-562.
  5. Dai Y, Vaught TD, Boone J, Chen SH, Phelps CJ, Ball S, Monahan JA, Jobst PM, McCreath KJ, Lamborn AE, Cowell-Lucero JL, Wells KD, Colman A, Polejaeva IA, Ayares DL (2002): Targeted disruption of the alpha1,3-galactosyltransferase gene in cloned pigs. Nat Biotechnol 20:251-255. https://doi.org/10.1038/nbt0302-251
  6. Phelps CJ, Koike C, Vaught TD, Boone J, Wells KD, Chen SH, Ball S, Specht SM, Polejaeva IA, Monahan JA, Jobst PM, Sharma SB, Lamborn AE, Garst AS, Moore M, Demetris AJ, Rudert WA, Bottino R, Bertera S, Trucco M, Starzl TE, Dai Y, Ayares DL (2003): Production of alpha 1,3-galactosyltransferase-deficient pigs. Science 299: 411-414. https://doi.org/10.1126/science.1078942
  7. Cooper DK, Dorling A, Pierson RN, 3rd, Rees M, Seebach J, Yazer M, Ohdan H, Awwad M, Ayares D (2007): Alpha 1,3-galactosyltransferase gene-knockout pigs for xenotransplantation: where do we go from here?. Transplantation 84:1-7. https://doi.org/10.1097/01.tp.0000260427.75804.f2
  8. Lin CC, Cooper DK, Dorling A (2009): Coagulation dysregulation as a barrier to xenotransplantation in the primate. Transpl Immunol 21:75-80. https://doi.org/10.1016/j.trim.2008.10.008
  9. Petersen B, Ramackers W, Tiede A, Lucas-Hahn A, Herrmann D, Barg-Kues B, Schuettler W, Friedrich L, Schwinzer R, Winkler M, Niemann H (2009): Pigs transgenic for human thrombomodulin have elevated production of activated protein C. Xenotransplantation 16:486-495. https://doi.org/10.1111/j.1399-3089.2009.00537.x
  10. Ekser B, Rigotti P, Gridelli B, Cooper DK (2009): Xenotransplantation of solid organs in the pig-to-primate model. Transpl Immunol 21:87-92. https://doi.org/10.1016/j.trim.2008.10.005
  11. Cowan PJ, Shinkel TA, Fisicaro N, Godwin JW, Bernabeu C, Almendro N, Rius C, Lonie AJ, Nottle MB, Wigley PL, Paizis K, Pearse MJ, d'Apice AJ (2003): Targeting gene expression to endothelium in transgenic animals: a comparison of the human ICAM-2, PECAM-1 and endoglin promoters. Xenotransplantation 10:223-231. https://doi.org/10.1034/j.1399-3089.2003.01140.x
  12. Godwin JW, Fisicaro N, d'Apice AJ, Cowan PJ (2006): Towards endothelial cell-specific transgene expression in pigs: characterization of the pig ICAM-2 promoter. Xenotransplantation 13:514-521. https://doi.org/10.1111/j.1399-3089.2006.00341.x
  13. Hellqvist M, Mahlapuu M, Blixt A, Enerback S, Carlsson P (1998): The human forkhead protein FREAC- 2 contains two functionally redundant activation domains and interacts with TBP and TFIIB. J Biol Chem 273:23335-23343. https://doi.org/10.1074/jbc.273.36.23335
  14. Cowan PJ, Tsang D, Pedic CM, Abbott LR, Shinkel TA, d'Apice AJ, Pearse MJ (1998): The human ICAM- 2 promoter is endothelial cell-specific in vitro and in vivo and contains critical Sp1 and GATA binding sites. J Biol Chem 27311737-27311744.
  15. Dai C, McAninch RE, Sutton RE (2004): Identification of synthetic endothelial cell-specific promoters by use of a high-throughput screen. J Virol 78:6209- 6221. https://doi.org/10.1128/JVI.78.12.6209-6221.2004
  16. Roeder RG (1996): The role of general initiation factors in transcription by RNA polymerase II. Trends Biochem Sci 21:327-335. https://doi.org/10.1016/S0968-0004(96)10050-5
  17. Nikolov DB, Burley SK (1997): RNA polymerase II transcription initiation: a structural view. Proc Natl Acad Sci USA 94:15-22. https://doi.org/10.1073/pnas.94.1.15
  18. Lee TI, Young RA (2000): Transcription of eukaryotic protein-coding genes. Annu Rev Genet 34:77-137. https://doi.org/10.1146/annurev.genet.34.1.77