Utility of DNA repair protein foci for the detection of putative BRCA1 pathway defects in breast cancer biopsies.
The DNA damage response pathway controlled by the breast cancer and Fanconi anemia (FA) genes can be disrupted by genetic or epigenetic mechanisms in breast cancer. Defects in this pathway may render the affected tumors hypersensitive to DNA-damaging agents. The identification of these defects poses a challenge because of the large number of genes involved in the FA/BRCA pathway. Many pathway components form subnuclear repair protein foci upon exposure to ionizing radiation in vitro, but it was unknown whether foci can be detected in live cancer tissues. Thus, the goal of this pilot study was to identify pathway defects by using a novel ex vivo foci biomarker assay on tumor biopsies. Fresh pretreatment biopsy specimens from patients with locally advanced sporadic breast cancer were irradiated or mock-treated in the laboratory (ex vivo). Foci formation of DNA repair proteins BRCA1, FANCD2, and RAD51 was detected by immunofluorescence microscopy. Three out of seven tumors showed intact radiation-induced foci formation, whereas the other four tumors exhibited a defective foci response. Notably, three of the foci-defective tumors were estrogen receptor/progesterone receptor/HER2-negative (triple-negative), a phenotype that has been associated with BRCA1 deficiency. In conclusion, in this pilot study, we report the successful detection of BRCA1, FANCD2, and RAD51 foci in breast cancer biopsies irradiated ex vivo. Our approach represents a potentially powerful biomarker assay for the detection of pre-existing and functionally important defects within the complex FA/BRCA pathway, which may ultimately allow us to tailor cancer treatment to the DNA repair profile of individual tumors.
Willers
Henning
H
Laboratory of Cellular and Molecular Radiation Oncology, Department of Radiation Oncology, Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, Massachusetts, USA.
Taghian
Alphonse G
AG
Luo
Chen-Mei
CM
Treszezamsky
Alejandro
A
Sgroi
Dennis C
DC
Powell
Simon N
SN
D019313:BRCA1 Protein / Q000172:deficiency / Q000378:metabolism*; D001706:Biopsy; D001940:Breast / Q000473:pathology*; D001943:Breast Neoplasms / Q000473:pathology*; D004260:DNA Repair / Q000528:radiation effects; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000378:metabolism; D005260:Female; D005629:Frozen Sections; D006801:Humans; D051135:Rad51 Recombinase / Q000378:metabolism; D011839:Radiation, Ionizing; D015398:Signal Transduction / Q000528:radiation effects
D019313:BRCA1 Protein; D052236:Fanconi Anemia Complementation Group D2 Protein; C495153:RAD51 protein, human; D051135:Rad51 Recombinase