title: VP22 enhances the expression of glucocerebrosidase in human Gaucher II fibroblast cells mediated by lentiviral vectors.
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.
authors.affiliation: Brain Tumor Research Center, Beijing Neurosurgical Institute, Beijing Tiantan Hospital affiliated to Capital Medical University, Tiantanxili 6, Chongwen District, Beijing, 100050, People's Republic of China.
authors.forename: Gui-shan
authors.initials: GS
authors.lastname: Jin
authors.forename: Gui-dong
authors.initials: GD
authors.lastname: Zhu
authors.forename: Zhi-gang
authors.initials: ZG
authors.lastname: Zhao
authors.forename: Fu-sheng
authors.initials: FS
authors.lastname: Liu
mesh_terms: D004789:Enzyme Activation; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005776:Gaucher Disease / Q000235:genetics / Q000628:therapy*; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005962:Glucosylceramidase / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D018259:Herpesvirus 1, Human / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D014162:Transfection; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D015678:Viral Structural Proteins; C073413:herpes simplex virus type 1 protein VP22; D005962:Glucosylceramidase