VP22 enhances the expression of glucocerebrosidase in human Gaucher II fibroblast cells mediated by lentiviral vectors

Clin Exp Med. 2012 Sep;12(3):135-43. doi: 10.1007/s10238-011-0152-7. Epub 2011 Aug 28.

Abstract

Gaucher disease is an autosomal recessive lysosomal storage disorder resulting in a deficiency of glucocerebrosidase (GC). Imiglucerase, a recombinant form of GC, has been successfully used in the treatment of Gaucher disease and has been shown to be a good potential candidate for gene therapy. However, its low transduction efficiency and short duration of expression have limited it as a gene therapy strategy. VP22, the herpes simplex virus type I tegument protein, is known to facilitate intercellular protein transport, thus making it a promising tool for improving gene transfer efficiency. To investigate whether the fusion of VP22 to GC could improve its therapeutic efficiency for Gaucher disease, the lentiviral vectors pHIV-GC and pHIV-VP(22)-GC were constructed and confirmed by PCR or RT-PCR. After packaging, the vectors were transduced into human Gaucher II fibroblast cells (GII cells). Flow cytometric analysis revealed that the GC expression rates in lenti-VP(22)-GC-transduced GII cells were higher by comparison than those in lenti-GC-transduced GII cells. A Western blot demonstrated higher levels of GC protein expression in lenti-VP(22)-GC-transduced GII cells. In addition, the long-term expression levels and increased GC activities in lenti-VP(22)-GC-transduced GII cells were also observed. These data implicate that VP22-mediated effects may be useful for enhancing the efficacy of this Gaucher disease treatment.

MeSH terms

  • Enzyme Activation
  • Fibroblasts / cytology
  • Fibroblasts / enzymology*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Gaucher Disease / genetics
  • Gaucher Disease / therapy*
  • Gene Expression Regulation, Enzymologic
  • Gene Transfer Techniques*
  • Genetic Vectors*
  • Glucosylceramidase / genetics
  • Glucosylceramidase / metabolism*
  • HEK293 Cells
  • Herpesvirus 1, Human / genetics
  • Humans
  • Lentivirus / genetics
  • Plasmids / genetics
  • Plasmids / metabolism
  • Primary Cell Culture
  • Transfection
  • Viral Structural Proteins / genetics
  • Viral Structural Proteins / metabolism*

Substances

  • Viral Structural Proteins
  • herpes simplex virus type 1 protein VP22
  • Glucosylceramidase