ERCC8

DNA excision repair protein ERCC-8 (Cockayne syndrome WD repeat protein CSA) [CKN1] [CSA]

Publications

Analysis of somatic mutations identifies signs of selection during in vitro aging of primary dermal fibroblasts.

PubMed

Full text

Somatic mutations are critical for cancer development and may play a role in age-related functional decline. Here, we used deep sequencing to analyze the prevalence of somatic mutations during in vitro cell aging. Primary dermal fibroblasts from healthy subjects of young and advanced age, from Hutchinson-Gilford progeria syndrome and from xeroderma pigmentosum complementation groups A and C, were first restricted in number and then expanded in vitro. DNA was obtained from cells pre- and post-expansion and sequenced at high depth (1656× mean coverage), over a cumulative 290 kb target region, including the exons of 44 aging-related genes. Allele frequencies of 58 somatic mutations differed between the pre- and post-cell culture expansion passages. Mathematical modeling revealed that the frequency change of three of the 58 mutations was unlikely to be explained by genetic drift alone, indicative of positive selection. Two of these three mutations, [[CDKN2A]] c.53C>T (T18M) and ERCC8 c.*772T>A, were identified in cells from a patient with XPA. The allele frequency of the [[CDKN2A]] mutation increased from 0% to 55.3% with increasing cell culture passage. The third mutation, BRCA2 c.6222C>T (H2074H), was identified in a sample from a healthy individual of advanced age. However, further validation of the three mutations suggests that other unmeasured variants probably provide the selective advantage in these cells. Our results reinforce the notions that somatic mutations occur during aging and that some are under positive selection, supporting the model of increased tissue heterogeneity with increased age.

MeSH Terms

• Adolescent
• Aged, 80 and over
• Cells, Cultured
• Cellular Senescence
• Child
• Child, Preschool
• DNA
• Female
• Fibroblasts
• Humans
• Male
• Mutation
• Sequence Analysis, RNA
• Skin

Keywords

• aging cell
• cell mosaicism
• genome instability
• molecular biology of aging
• positive selection
• somatic mutation
• tissue heterogeneity