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Naked DNA expressing two isoforms of hepatocyte growth factor induces collateral artery augmentation in a rabbit model of limb ischemia

Abstract

Hepatocyte growth factor (HGF) has been shown to induce angiogenesis in vivo and has potential as a candidate gene for ‘therapeutic angiogenesis’. In vivo, two isoforms of HGF, HGF723 and HGF728, consisting of 723 and 728 amino acids, are generated through alternative splicing between exons 4 and 5, but the biological effects of their coexpression have not yet been elucidated. In this study, we generated a series of genomic–complementary DNA (cDNA) hybrids of the HGF gene by inserting various truncated intron 4 into the junction of exons 4 and 5 of HGF cDNA and analyzed the biological activities of these hybrid constructs. We showed that: (1) the hybrid called HGF-X7, which contained 1502 base pairs of intron 4, could drive a higher level of HGF expression than other hybrid constructs and cDNAs of each isoform alone; (2) the pCK vector was most efficient for the gene expression of HGF-X7; (3) coexpression of both isoforms of HGF could more efficiently induce the migration of human umbilical vein endothelial cell (HUVEC) and of the mouse myoblast cell line C2C12 myoblasts than a single isoform of HGF and human vascular endothelial growth factor (VEGF)165 at a given concentration; (4) intramuscular administration of pCK-HGF-X7 resulted in transient and localized HGF expression in the injected muscle without an increase in the HGF protein levels in other tissues including serum; and (5) intramuscular injection of pCK-HGF-X7 could more efficiently increase the number of angiographically recognizable collateral vessels, as well as improve an intra-arterial Doppler wire-measured blood flow in the rabbit model of hindlimb ischemia when compared with the identical vector encoding VEGF165 gene. These results showed that transfer of the genomic–cDNA hybrid of the HGF gene could be used as a potential therapeutic approach to human vascular diseases.

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Acknowledgements

This work was supported by grants from the Korean Ministry of Knowledge Economy (grant no. 10031644) and also from the Science Research Center program of the Korea Science and Engineering Foundation (grant no. R11-2005-009-06003-0).

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Correspondence to S Kim.

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Competing interests

Woong Hahn, Dong-Sik Kim, Won-Sun Yoo, Sung-Dong Lee, Jung-Hee Won, Jong-Mook Kim and Sunyoung Kim are employees or shareholders of ViroMed Co., Ltd., whose potential product was studied in this work. Wook-Bum Pyun is the medical advisor of ViroMed Co., Ltd. The other authors declare no conflict of interest.

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Pyun, WB., Hahn, W., Kim, DS. et al. Naked DNA expressing two isoforms of hepatocyte growth factor induces collateral artery augmentation in a rabbit model of limb ischemia. Gene Ther 17, 1442–1452 (2010). https://doi.org/10.1038/gt.2010.101

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  • DOI: https://doi.org/10.1038/gt.2010.101

Keywords

  • hepatocyte growth factor (HGF)
  • isoform
  • ischemic limb disease

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