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Article Dans Une Revue Oncogene Année : 2022

Microsatellite instability detection in breast cancer using drop-off droplet digital PCR

Shufang Renault
  • Fonction : Auteur correspondant

Résumé

The use of conventional methods (immunohistochemistry, pentaplex PCR) for detecting microsatellite instability (MSI), a predictive biomarker of immunotherapy efficacy, is debated for cancers with low MSI prevalence, such as breast cancer (BC). We developed two multiplex drop-off droplet digital PCR (ddPCR) assays targeting four microsatellites, initially identified from public BC whole-genome sequencing dataset. Performances of the assays were investigated and 352 tumor DNA and 28 circulating cell-free DNA from BC patients, with unknown MSI status were blindly screened. Cross-validation of ddPCR MSI status with other MSI detection methods was performed. We then monitored circulating tumor DNA (ctDNA) dynamics before and during pembrolizumab immunotherapy in one patient with MSI-high (MSI-H) metastatic BC. The assays showed high analytical specificity and sensitivity (limit of detection = 0.16%). Among N = 380 samples, seven (1.8%) were found as MSI-H by ddPCR with six of them confirmed by next-generation sequencing (NGS). Specificity was 100% in N = 133 microsatellite stable BC submitted to NGS. In the patient with MSI-H metastatic BC, ctDNA monitoring revealed an early decrease of microsatellite mutant allelic frequencies during immunotherapy. These results demonstrated MSI detection by ddPCR, a non-invasive, fast and cost-effective approach, allowing for large pre-screening of BC patients who may benefit from immunotherapy.
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Dates et versions

hal-03949622 , version 1 (20-01-2023)

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Khadidja Zeyneb Klouch, Marc Henri Stern, Olfa Trabelsi-Grati, Nicolas Kiavue, Luc Cabel, et al.. Microsatellite instability detection in breast cancer using drop-off droplet digital PCR. Oncogene, 2022, 41 (49), pp.5289-5297. ⟨10.1038/s41388-022-02504-6⟩. ⟨hal-03949622⟩
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