Enhancing 1 alpha-hydroxylase activity with the 25-hydroxyvitamin D-1 alpha-hydroxylase gene in cultured human keratinocytes and mouse skin.
1 alpha,25-Dihydroxyvitamin D(3) (1 alpha,25(OH(2))D(3)) and its analogs are used to treat psoriasis because of their potent antiproliferative activity. They have the potential for causing hypercalcemia, however, and patients often become resistant to the drug. We examined the feasibility of enhancing the cutaneous production of 1 alpha,25(OH(2))D(3) using a human 25-hydroxyvitamin D-1 alpha-hydroxylase (1 alpha-OHase) plasmid. The 1 alpha-OHase gene was fused to the green fluorescent protein gene (1 alpha-OHase-GFP) driven by the cytomegalovirus promoter. Transfection of cultured normal human keratinocytes with the 1 alpha-OHase-GFP plasmid resulted in a marked increase in the expression of 1 alpha-OHase-GFP in the mitochondria. Transfection of keratinocytes with 1 alpha-OHase-GFP or 1 alpha-OHase plasmids in vitro enhanced the 1 alpha-OHase activity substantially and increased the sensitivity of the keratinocytes to the antiproliferative effect of 25(OH)D(3). The 1 alpha-OHase-GFP plasmid was topically applied to shaved C57/BL6 mice. Twenty-four hours after topical application, immunohistochemical analysis of the skin for 1 alpha-OHase-GFP revealed the presence of 1 alpha-OHase-GFP in the epidermis and epidermal appendages including the hair follicles. The results from this study offer a unique new approach for the topical treatment of hyperproliferative disorders such as psoriasis and skin cancer using the 1 alpha-OHase gene that could locally increase the production of 1 alpha,25(OH(2))D(3) without causing hypercalcemia or resistance.
Flanagan
J N
JN
Vitamin D, Skin and Bone Research Laboratory, Section of Endocrinology, Nutrition, and Diabetes, Department of Medicine, Boston University Medical Center, Boston, Massachusetts, USA.
Whitlatch
L W
LW
Chen
T C
TC
Zhu
X H
XH
Holick
M T
MT
Kong
X F
XF
Holick
M F
MF
D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002112:Calcifediol / Q000378:metabolism; D002117:Calcitriol / Q000096:biosynthesis; D002478:Cells, Cultured; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D015603:Keratinocytes / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids; D011565:Psoriasis / Q000628:therapy; D012867:Skin / Q000201:enzymology*; D014162:Transfection
D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase; D002117:Calcitriol; D002112:Calcifediol