A lentiviral vector-based genetic sensor system for comparative analysis of permeability and activity of vitamin D3 analogues in xenotransplanted human skin

Exp Dermatol. 2013 Mar;22(3):178-83. doi: 10.1111/exd.12091.

Abstract

Vitamin D3 analogues are widely used topical and oral remedies for various ailments such as psoriasis, osteoporosis and secondary hyperparathyroidism. In topical treatment, high skin permeability and cellular uptake are key criteria for beneficial effects due to the natural barrier properties of skin. In this study, we wish to establish an in vivo model that allows the comparison of permeability and activity of vitamin D3 analogues in human skin. We generate a bipartite, genetic sensor technology that combines efficient lentivirus-directed gene delivery to xenotransplanted human skin with vitamin D3-induced expression of a luciferase reporter gene and live imaging of animals by bioluminescence imaging. Based on the induction of a transcriptional activator consisting of the vitamin D receptor fused to the Gal4 DNA-binding domain, the vitamin D3-responsive sensor facilitates non-invasive and rapid assessment of permeability and functional properties of vitamin D3 analogues. By topical application of a panel of vitamin D3 analogues onto 'sensorized' human skin, the sensor produces a drug-induced readout with a magnitude and persistence that allow a direct comparative analysis of different analogues. This novel genetic tool has great potential as a non-invasive in vivo screening system for further development and refinement of vitamin D3 analogues.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Topical
  • Animals
  • Cell Line
  • Cell Membrane Permeability / physiology*
  • Cholecalciferol / administration & dosage
  • Cholecalciferol / analogs & derivatives*
  • Cholecalciferol / metabolism*
  • Genetic Vectors / genetics*
  • HEK293 Cells
  • Humans
  • Keratinocytes / cytology
  • Keratinocytes / metabolism
  • Lentivirus / genetics*
  • Luciferases / genetics
  • Luciferases / metabolism
  • Luminescent Measurements
  • Mice
  • Mice, SCID
  • Models, Animal
  • Skin / cytology
  • Skin / metabolism*
  • Skin Transplantation
  • Transplantation, Heterologous*

Substances

  • Cholecalciferol
  • Luciferases