Systemic delivery of secreted protein by grafts of epidermal keratinocytes: prospects for keratinocyte gene therapy.
Grafts of autologous keratinocytes genetically altered to secrete a new gene product are a potential vehicle for gene therapy. To consider the feasibility of such an approach, we have examined the ability of keratinocytes to secrete and deliver apolipoprotein E (apoE) to the circulation of mice bearing grafts of human keratinocytes. The grafted keratinocytes secreted two forms of apoE, an endogenous apoE encoded in the genome and a recombinant apoE encoded in a transfected gene construct. In vitro studies showed that endogenous apoE was secreted from basal keratinocytes whereas recombinant apoE was secreted from basal as well as suprabasal cells. On the basis of amounts of recombinant apoE present in the serum of grafted mice, we estimate that a graft occupying 2% of the surface area of an adult human would deliver 6.5-8.3 mg of recombinant apoE protein per day.
Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York, Stony Brook 11794-8702.
E S
ES
Fenjves
J
J
Smith
S
S
Zaradic
L B
LB
Taichman
D000818:Animals; D001057:Apolipoproteins E / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002478:Cells, Cultured; D015316:Genetic Therapy; D006207:Half-Life; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000637:transplantation*; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
D001057:Apolipoproteins E; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D011993:Recombinant Fusion Proteins