title: Characterization of transgenic mice with the expression of phenylalanine hydroxylase and GTP cyclohydrolase I in the skin.
abstract: Phenylketonuria (PKU) is a metabolic disease causing increased levels of phenylalanine in blood and body fluids. Circulating phenylalanine is normally cleared by phenylalanine hydroxylase (PAH) expressed in the liver. The aim of this study is to exploit the skin as a 'metabolic sink' removing phenylalanine from the blood. We have previously showed that the overexpression of PAH and GTP cyclohydrolase I (GTP-CH), the rate-limiting enzyme in the synthesis of the cofactor for PAH, leads to high levels of phenylalanine clearance in primary human keratinocytes. In this study, we have investigated the 'metabolic sink' strategy in an in vivo model by developing three lines of transgenic mice expressing PAH and GTP-CH in various layers of the skin. The promoters used were keratin 14 (K14), involucrin (INV) and a truncated variant of Keratin 1 (K1). The mice were crossbred to a mouse model of human PKU, the PAH(enu2) mouse, in order to obtain mice that do not express PAH in the liver and the kidney. Transgenic mice containing the INV and K14 promoters expressed PAH and GTP-CH in the epidermis. However, the K1 promoter did not lead to detectable gene expression. Analysis of the mice showed that no phenotypic effect was observed in mice expressing PAH and GTP-CH from the INV promoter. However, low level of phenylalanine clearance was observed in mice expressing PAH and GTP-CH from the K14 promoter, suggesting that the skin can be genetically engineered to function as a 'metabolic sink'.
authors.affiliation: Department of Human Genetics, University of Aarhus, Aarhus, Denmark. rikkec@humgen.au.dk
authors.forename: Rikke
authors.initials: R
authors.lastname: Christensen
authors.forename: Leena
authors.initials: L
authors.lastname: Alhonen
authors.forename: Jarmo
authors.initials: J
authors.lastname: Wahlfors
authors.forename: Maria
authors.initials: M
authors.lastname: Jakobsen
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Jensen
mesh_terms: D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004195:Disease Models, Animal; D006136:GTP Cyclohydrolase / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy; D006801:Humans; D053547:Keratin-14; D007633:Keratins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D010641:Phenotype; D010649:Phenylalanine / Q000378:metabolism; D010651:Phenylalanine Hydroxylase / Q000235:genetics* / Q000378:metabolism*; D010661:Phenylketonurias / Q000201:enzymology / Q000235:genetics / Q000628:therapy; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000235:genetics; D012867:Skin / Q000201:enzymology* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; C507540:KRT14 protein, human; D053547:Keratin-14; C507446:Krt14 protein, mouse; D011498:Protein Precursors; D010649:Phenylalanine; C032253:involucrin; D007633:Keratins; D010651:Phenylalanine Hydroxylase; D006136:GTP Cyclohydrolase