Abstract
Hutchinson-Gilford progeria syndrome (HGPS; MIM 176670) is a rare disease characterized by accelerated aging. In this study, light and immunofluorescence microscopy were used to assess morphological changes, measures of cell growth kinetics and gene expression profiles in HGPS cells and normal fibroblasts in culture. A filtering strategy was developed based on differentially expressed transcripts seen consistently across three culture stages based on cell passage number. This filtering strategy produced a list of 66 unique differentially expressed genes, of which ∼40% were upregulated in HGPS cells compared to normal fibroblasts. The increased mRNA expression in HGPS cells that was seen for one gene defined using this strategy—namely ANK3— was validated using quantitative reverse-transcriptase amplification, Western analysis and immunofluorescence microscopy, all of which showed significantly increased ankyrin G expression. These findings demonstrate differences in morphology, growth kinetics and mRNA expression profiles in HGPS cells compared to normal fibroblasts in culture, including increased expression of ANK3/ankyrin G. Furthermore, other genes that co-clustered with ANK3 might provide mechanistic clues regarding senescence in cultured HGPS cells.
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Acknowledgments
R.A.H. is supported by the Jacob J. Wolfe Distinguished Medical Research Chair, the Edith Schulich Vinet Canada Research Chair (Tier I) in Human Genetics, a Career Investigator award from the Heart and Stroke Foundation of Ontario. J.G.P. is supported by a Career Investigator award from the Heart and Stroke Foundation of Ontario. Operating support came from the Canadian Institutes for Health Research, the Heart and Stroke Foundation of Ontario (PRG4854), the Ontario Research and Development Challenge Fund (Project #0507) and by Genome Canada. C.M.W. was the recipient of a summer student award from the Ontario Genomics Institute.
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Wang, J., Robinson, J.F., O’Neil, C.H. et al. Ankyrin G overexpression in Hutchinson-Gilford progeria syndrome fibroblasts identified through biological filtering of expression profiles. J Hum Genet 51, 934–942 (2006). https://doi.org/10.1007/s10038-006-0042-0
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DOI: https://doi.org/10.1007/s10038-006-0042-0
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