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

• Published in: Gene therapy Vol. 17; no. 12; pp. 1442 - 1452
• Main Authors: Pyun, W-B, Hahn, W, Kim, D-S, Yoo, W-S, Lee, S-D, Won, J-H, Rho, B-S, Park, Z-Y, Kim, J-M, Kim, S
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-12-2010; Nature Publishing Group
• Subjects: 631/443/592/16; 631/443/592/1939; 631/45/127/1211; 631/61/51/201; Alternative splicing; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Angiogenesis; Animals; Applied cell therapy and gene therapy; Arteries - growth & development; Arteries - metabolism; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Biotechnology; Blood flow; Care and treatment; Cell Biology; Cell Line; Cell Movement - drug effects; Collateral Circulation - drug effects; Complementary DNA; Deoxyribonucleic acid; Disease Models, Animal; DNA; DNA - genetics; DNA - therapeutic use; DNA, Complementary - genetics; Endothelial cells; Exons; Extremities - blood supply; Female; Fundamental and applied biological sciences. Psychology; Gene Expression; Gene Therapy; Gene Transfer Techniques; Genetic aspects; Genetic Engineering; Genetic Therapy; Genetic Vectors - genetics; Genomics; Growth factors; Health aspects; Health. Pharmaceutical industry; Hepatocyte growth factor; Hepatocyte Growth Factor - genetics; Hepatocyte Growth Factor - metabolism; Hepatocyte Growth Factor - pharmacology; HGF gene; Human Genetics; Humans; Hybrids; Industrial applications and implications. Economical aspects; Introns - genetics; Ischemia; Ischemia - physiopathology; Ischemia - therapy; Isoforms; Male; Medical sciences; Mice; Mice, Inbred BALB C; Myoblasts; Nanotechnology; original-article; Physiological aspects; Protein Isoforms - genetics; Protein Isoforms - metabolism; Rabbits; Regional Blood Flow - drug effects; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Umbilical vein; Vascular diseases; Vascular endothelial growth factor; South Korea; hepatocyte growth factor (HGF); ischemic limb disease; isoform; Molecular form; Hepatocyte growth factor; Rabbit; Cardiovascular disease; Lagomorpha; Artery; Vertebrata; Mammalia; Ischemia; Blood vessel; DNA; Limb; Models; Circulatory system; Gene therapy
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2010.101

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, HGF
723
and HGF
728
, 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 C
2
C
12
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
VEGF
165
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.