Telomerase expression restores dermal integrity to in vitro-aged fibroblasts in a reconstituted skin model.
The lifespan of human fibroblasts and other primary cell strains can be extended by expression of the telomerase catalytic subunit (hTERT). Since replicative senescence is accompanied by substantial alterations in gene expression, we evaluated characteristics of in vitro-aged dermal fibroblast populations before and after immortalization with telomerase. The biological behavior of these populations was assessed by incorporation into reconstituted human skin. Reminiscent of skin in the elderly, we observed increased fragility and subepidermal blistering with increased passage number of dermal fibroblasts, but the expression of telomerase in late passage populations restored the normal nonblistering phenotype. DNA microarray analysis showed that senescent fibroblasts express reduced levels of collagen I and III, as well as increased levels of a series of markers associated with the destruction of dermal matrix and inflammatory processes, and that the expression of telomerase results in mRNA expression patterns that are substantially similar to early passage cells. Thus, telomerase activity not only confers replicative immortality to skin fibroblasts, but can also prevent or reverse the loss of biological function seen in senescent cell populations.
Funk
W D
WD
Geron Corporation, 230 Constitution Drive, Menlo Park, California 94025, USA. wfunk@geron.com
Wang
C K
CK
Shelton
D N
DN
Harley
C B
CB
Pagon
G D
GD
Hoeffler
W K
WK
D020134:Catalytic Domain; D002460:Cell Line; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology*; D004268:DNA-Binding Proteins; D020405:Dermis / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology; D008954:Models, Biological; D010812:Physical Stimulation; D012313:RNA; D012879:Skin Physiological Phenomena; D019098:Telomerase / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
D004268:DNA-Binding Proteins; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase