Hyaluronidase and collagenase increase the transfection efficiency of gene electrotransfer in various murine tumors

Hum Gene Ther. 2012 Jan;23(1):128-37. doi: 10.1089/hum.2011.073. Epub 2011 Sep 9.

Abstract

One of the applications of electroporation/electropulsation in biomedicine is gene electrotransfer, the wider use of which is hindered by low transfection efficiency in vivo compared with viral vectors. The aim of our study was to determine whether modulation of the extracellular matrix in solid tumors, using collagenase and hyaluronidase, could increase the transfection efficiency of gene electrotransfer in histologically different solid subcutaneous tumors in mice. Tumors were treated with enzymes before electrotransfer of plasmid DNA encoding either green fluorescent protein or luciferase. Transfection efficiency was determined 3, 9, and 15 days posttransfection. We demonstrated that pretreatment of tumors with a combination of enzymes significantly increased the transfection efficiency of electrotransfer in tumors with a high extracellular matrix area (LPB fibrosarcoma). In tumors with a smaller extracellular matrix area and less organized collagen lattice, the increase was not so pronounced (SA-1 fibrosarcoma and EAT carcinoma), whereas in B16 melanoma, in which only traces of collagen are present, pretreatment of tumors with hyaluronidase alone was more efficient than pretreatment with both enzymes. In conclusion, our results suggest that modification of the extracellular matrix could improve distribution of plasmid DNA in solid subcutaneous tumors, demonstrated by an increase in transfection efficiency, and thus have important clinical implications for electrogene therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Collagen / metabolism
  • Collagenases / pharmacology*
  • Electroporation / methods*
  • Extracellular Matrix / metabolism
  • Extracellular Matrix / pathology
  • Female
  • Fibrosarcoma / metabolism
  • Fibrosarcoma / pathology
  • Fibrosarcoma / therapy
  • Gene Transfer Techniques*
  • Genetic Therapy / methods
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Hyaluronoglucosaminidase / pharmacology*
  • Luciferases / genetics
  • Luciferases / metabolism
  • Melanoma, Experimental / metabolism
  • Melanoma, Experimental / pathology
  • Melanoma, Experimental / therapy
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred CBA
  • Plasmids / administration & dosage
  • Plasmids / genetics
  • Plasmids / metabolism
  • Time Factors
  • Transfection

Substances

  • Green Fluorescent Proteins
  • Collagen
  • Luciferases
  • Hyaluronoglucosaminidase
  • Collagenases