Antisense oligonucleotide targeting fibroblast growth factor receptor (FGFR)‐1 stimulates cellular activity of hair follicles in an in vitro organ culture system

Abstract

The hair cycle is regulated by various molecules, among which FGF-5 has been shown to promote the transition from anagen to catagen. The FGFR-1, a trans-membrane receptor of FGF-5 with tyrosine kinase activity, is localized in the follicular papilla of hair follicles. In order to apply the antisense oligonucleotides targeting FGFR-1 as a treatment for baldness, we examined the effect of the oligonucleotides on hair follicle growth in a serum-free organ culture system. Vibrissal hair follicles from C3H/He mice were cultured in the presence of a reagent at 31 degrees C in 95% O(2)-5% CO(2) for 72 h. A 20-mer antisense nucleotide and its randomly arranged counterpart were prepared by predicting the effective target site of FGFR-1 mRNA. Cellular activity in the hair bulb was estimated by measuring the fluorescence intensity (FI) of the medium after incubation with AlamarBlue dye. The addition of 30 microM of the phosphorothioate form of antisense oligonucleotide (A1561TS) to the media increased the FI by 30%, whereas the control produced no detectable change. This effect was reproducible dose-dependent with maximal stimulation at 30 microM. Incorporation of the oligonucleotide into the follicular papilla was histologically confirmed by incubation with FITC-labeled phosphorothioate oligonucleotides, and the intact morphological structure of the hair bulb was maintained intact after a 72-h incubation. These results suggest the clinical utility of antisense nucleotide targeting FGFR-1 as a treatment for baldness.