Work

Characterization of an aFGF Gene Expression Vector with Therapeutic Potential

Year: 1998

Type: article

Abstract: ulatory concentrations of any growth factor is required Background.Topical application of growth factors to produce maximal cell proliferation [4].Somatic gene to wounds has proven to be suboptimal in achieving therapy describes the introduction and expression of epithelial growth and accelerating healing.We prorecombinant genes in somatic cells [1]; this technique pose transfection of fibroblasts with a gene for acidic could potentially deliver therapeutic doses of growth fibroblast growth factor (aFGF) which will allow confactors to wounds or injured gastrointestinal tissue in tinuous, local delivery of the growth factor to wounds, a continuous manner.ulcerative lesions, or healing tissues.Fibroblast growth factors (FGFs) have long been con-Methods.We utilized a pMEXneo vector containing sidered to be key mitogens, chemoattractants and reguthe human aFGF gene with a secretory signal selatory proteins in the wound area.Acidic fibroblast quence from the hst/KS3 gene to obtain continuous segrowth factor (aFGF) is one of nine homologous FGFs cretion of therapeutic doses of aFGF.NIH 3T3 fibrowhich stimulate cells of mesodermal and ectodermal blasts were transfected using a liposomal transfection origin [6].Acidic FGF, produced by fibroblasts, endoreagent and grown in selective media.thelial cells, macrophages, and platelets, is a potent Results.Dot blot hybridization with labeled complemitogen for wound fibroblasts and vascular endothelial mentary DNA probes revealed the presence of plasmid cells inducing new blood vessel growth [6] and has been DNA in transfected but not wild type fibroblasts.Intrademonstrated to accelerate healing of experimental cellular concentrations of aFGF remained low in gastric ulcerations [7].Unlike most polypeptide growth transfected cells; however, the media contained high factors, aFGF does not possess a secretory signal selevels (32 { 7 nM) of aFGF as measured by ELISA.Concentrations of aFGF capable of stimulating cell prolif-quence [8].Thus it is produced, bound to large moleeration were maintained for several weeks.cules such as heparan sulfate, but not released from the Conclusions.The aFGF cDNA was transcribed and cell in a soluble form.Proteases and endoglycosidases translated into a functional polypeptide that is seresponding to endothelial cell damage are likely to be creted from NIH 3T3 cells at physiologically significant responsible for cellular release of aFGF [9].Tissue levconcentrations.Stable transfection with a eukaryotic els of aFGF are seldom reported due in part to its short vector which induces secretion of aFGF at levels prohalf-life and heparan binding.The presence of aFGF moting cell growth holds promise for clinical applicain healing wounds is usually assessed by immunohistotion in wounds or healing tissue.Transfection could chemistry or Northern blot detection of mRNA.be achieved by topical or endoscopic injection of this NIH 3T3 murine fibroblasts were transfected with a type of vector.᭧ 1998 Academic Press eukaryotic expression vector containing an aFGF gene Key Words: fibroblast growth factor; plasmid; transcoupled to a signal peptide sequence to obtain secretion fection; wound healing.of this otherwise nonsecreted growth factor.We utilized a construct of pMEXneo expression vector containing a human aFGF coding frame.The signal se-

Source: Journal of Surgical Research

DOI: 10.1006/jsre.1998.5351