BRCA1 and BRCA2

Mutations in Breast Cancer

• The breast cancer susceptibility genes 1 and 2 (BRCA1 and BRCA2) are
important tumor suppressors that code for proteins involved in cell
regulation and the error-free repair of double-strand breaks (DSBs) in
DNA.
• Mutations in BRCA1 and/or BRCA2 result in the inability to effectively
repair DSBs. As a result, DSBs persist or are repaired via the error-
prone non-homologous end joining (NHEJ) mechanism, resulting in
genomic instability, accumulation of genetic alterations, and
chromosomal abnormalities leading to carcinogenesis.
• BRCA1 and BRCA2 mutations were the first homologous
recombination repair (HRR) gene mutations discovered to be linked to
breast and ovarian cancers.
• Mutations in BRCA1 and BRCA2 are associated with a high lifetime
risk of developing breast cancer.
• Historically, testing was used to identify BRCA mutations in
unaffected family members to potentially provide an opportunity to
take preventative measures.
• Today, BRCA1 and BRCA2 mutation status can also inform treatment
decisions.
Diagnostic Assays for BRCA1 and BRCA2
Testing
• While Sanger sequencing is considered the gold standard for
detection of genetic mutations, several next-generation sequencing
platforms are available for the screening and identification of BRCA1
and BRCA2 mutations.
• Testing should include full gene sequencing, deletion/duplication
analysis, and detection of known pathogenic/likely pathogenic
variants in a Clinical Laboratory Improvement Amendments (CLIA)-
certified and/or College of American Pathologists (CAP)-accredited
genetic testing laboratory.
How to Interpret BRCA1 and BRCA2 Test
Results
• Sequence variations in BRCA1 and BRCA2 fall into 3 broad categories,
single-nucleotide changes, small insertion or deletion events (indels), and
large genomic rearrangements (LGRs).
• Single-nucleotide changes and indels can be detected by direct (Sanger)
sequencing or next-generation sequencing.
• LGRs cannot be detected by direct (Sanger) sequencing. Instead,
alternative polymerase chain reaction (PCR)–based techniques (eg,
quantitative PCR, multiplex ligation-dependent probe amplification) are
required.
• Whether performing in-house testing or sending to a testing lab, ensure
the methodology is comprehensive and includes LGRs and indels.
Proper Interpretation of Test Results Is
Critical for Informing Treatment Decisions
• Many different mutations are possible in the BRCA genes; therefore,
test results may not be a simple positive or negative.