> Details

Sheridan, Paul J.; Lawrie, Allan; Crossman, David C.; Holt, Cathy M.; Newman, Christopher M.

VP22‐mediated intercellular transport correlates with enhanced biological activity of MybEngrailed but not (HSV‐I) thymidine kinase fusion proteins in primary vascular cells following non‐viral transfection

Reference
management

Bookmarks

Remove from
bookmarks

Share this on Whatsapp

Close

Bookmarks



You can manage bookmarks using lists, please log in to your user account for this.
  • Media type: E-Article
  • Title: VP22‐mediated intercellular transport correlates with enhanced biological activity of MybEngrailed but not (HSV‐I) thymidine kinase fusion proteins in primary vascular cells following non‐viral transfection
  • Contributor: Sheridan, Paul J.; Lawrie, Allan; Crossman, David C.; Holt, Cathy M.; Newman, Christopher M.
  • Published: Wiley, 2005
  • Published in: The Journal of Gene Medicine, 7 (2005) 3, Seite 375-385
  • Language: English
  • DOI: 10.1002/jgm.679
  • ISSN: 1099-498X; 1521-2254
  • Keywords: Genetics (clinical) ; Drug Discovery ; Genetics ; Molecular Biology ; Molecular Medicine
  • Origination:
  • Footnote:
  • Description: <jats:title>Abstract</jats:title><jats:sec><jats:title>Background</jats:title><jats:p>The intercellular transport properties of the herpes simplex virus (HSV) protein VP22 have been harnessed to enhance the effectiveness of viral gene transfer. We investigated the intercellular transport and biological effects of VP22 fused with the dominant negative c‐Myb chimera, MybEngrailed (MybEn) and HSV‐I thymidine kinase (TK), in primary vascular smooth muscle cells (VSMC) following non‐viral transfection.</jats:p></jats:sec><jats:sec><jats:title>Materials and methods</jats:title><jats:p>Porcine VSMC transfected with plasmids encoding MybEn, TK and their respective N‐ and C‐terminal VP22 fusion proteins were assayed for the extent and distribution of transgene expression (by immunohistochemistry), culture growth and apoptosis.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>The N‐terminal MybEn fusion with VP22 (MybEnVP22) and both TK fusions, but not VP22MybEn, exhibited intercellular spread from primary transfected to up to 200 surrounding cells. <jats:italic>pMybEnVP22</jats:italic>‐transfected cultures exhibited growth inhibition and apoptosis rates that were 10.6 ± 3.6 and 3.2 ± 1.0 fold higher than in <jats:italic>pMybEn</jats:italic>‐transfected cultures; <jats:italic>pVP22MybEn</jats:italic>‐transfected cultures showed no difference in these parameters. <jats:italic>pTK</jats:italic>‐transfected cultures underwent 60–70% cell death in the presence of ganciclovir despite &lt;2% primary transfection, which was not increased in cultures transfected with plasmids encoding VP22‐TK fusions.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>The close correlation between immunocytochemical and biological assays suggests that intercellular transport is crucial to the enhanced biological activity of the MybEnVP22 fusion. The ‘intrinsic’ bystander activity of TK was 4‐fold greater than was ‘engineered’ by VP22 fusion, probably reflecting the abundance of gap junctions between VSMC. VP22 fusion may enhance the efficiency of non‐viral gene delivery when combined with the appropriate therapeutic transgene, target tissue and transfection method. Copyright © 2004 John Wiley &amp; Sons, Ltd.</jats:p></jats:sec>