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Biology subjects

LaBarge, M.

Publications and source records attributed to LaBarge, M..

2 recordsLinked to original sources

The Breakome of BRCA1 and BRCA2 Pathway Mutation Carriers Reveals Early Processes in Breast Oncogenesis

DNA double-strand breaks (DSBs) can lead to genomic instability in cancer. Cells rely on an efficient DNA damage response (DDR) to maintain their DNA integrity and prevent oncogenic transformation. However, the early events that connect recurrent DNA damage to oncogenesis are not yet fully understood. Here, using next generation sequencing we comprehensively surveyed genomes to identify DSBs in primary cells of non-malignant carriers of BRCA1 and BRCA2 mutations (BRCAmut), categorized as high-risk patients, to characterize the effects of homologous recombination (HR) loss on cancer initiation. We demonstrate that the landscape of physiological DSBs in BRCAmut mammary epithelial cells differs from that of healthy controls and resemble more the DSB pattern observed in breast cancer cells. Our results reveal that proto-oncogenes and tumor suppressors contain more breaks in BRCAmut samples, and that genes with a high number of DSBs tend to be more highly expressed. These genes containing a high number of DSBs are also often mutated in breast cancer tumors. Finally, genes with high DSBs in mammary epithelial cells from women with BRCAmut exhibit a strong correlation with homologous recombination repair. Together, our findings underscore the impact of BRCA loss on the early stages of carcinogenesis and highlight future possibilities for early cancer detection. Graphical abstractWhen BRCA is intact, genes that are highly broken are properly repaired via HR, preserving DNA integrity. When BRCA is mutant, impairing its function, highly broken transcriptional DSB genes emerge, no longer able to be efficiently repaired via HR, and are found at genes related to cancer signaling. Breakome of enriched breaks at high-risk model resembles breast cancer breakome, and breaks can be found in genes known to be frequently mutated in breast cancer.

cancer biology↗

Microenvironment-contextual cell signaling is attenuated with age

Post-menopausal women are more prone to breast cancer than younger women. The increased frequency of age-related breast cancers is likely due to interactions between acquired mutations and age-dependent epigenetic changes that affect mammary epithelial lineage fidelity. We hypothesized that the aging process fundamentally affects how human mammary epithelial cells (HMEC) respond to microenvironmental signals, resulting in increased susceptibility to oncogenic transformation. In order to measure microenvironmental cell signaling in normal finite lifespan HMEC, we applied a novel microsphere-based flow cytometry technology. The microsphere cytometry allows multiparametric single cell quantification of signaling pathway activity and lineage-specific marker expression in cells adhered to surface-functionalized microspheres that mimic specific microenvironments. Using this approach, we analyzed age-dependent changes in human mammary myoepithelial and luminal epithelial cells exposed to different ECM and growth factors. We found that ECM-mediated MAP kinase and PI3 kinase activation levels in HMEC were attenuated with age. Older luminal cells displayed higher surface integrin levels consistent with acquired basal identity, albeit with decreased integrin activation and increased Src-signaling relative to myoepithelial cells. We show that the diminished signaling magnitude in HMEC from older women correlated with reduced probability of activating oncogene-induced senescence. We propose that age-related changes in ECM-mediated epithelial cell regulation may impair protective tumor suppression mechanisms and increase breast cancer susceptibility.

cell biology↗