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• W3041299573 abstract "Pancreatic ductal adenocarcinoma (PDAC) is associated with poor prognosis and high mortality worldwide. At the time of diagnosis 80% of cases are already locally advanced or metastatic. Despite the emergence of new therapies for the treatment of metastatic or locally advanced PDAC, reliable prognostic/predictive biomarkers are lacking. PDAC harbours mutations in the KRAS gene on exons 2 and 3, in 80-90% of patients. The aim of the present study was the detection and quantification of RAS mutations in the ct DNA (circulating tumour DNA) of patients with inoperable PDAC by BEAMing digital polymerase chain reaction (PCR) in order to (a) evaluate the correlation with tissue RAS mutation status in the tumour, (b) study the baseline prognostic significance, and (c) study the predictive significance of ctDNA RAS mutation kinetics for the prediction of response to systemic/neoadjuvant therapy, in correlation with Ca 19-9 serum tumor marker and the CT scans response. The plasma samples were collected at two time points, the first one at diagnosis, before chemotherapy and the second one 4-6 weeks later. BEAMing digital PCR technique (KRAS/NRAS profiling-Digital PCR-OncoBEAMTM RAS CRC kit-Sysmex Inostics) is characterized by high sensitivity and allows the quantification of plasma mutant alleles. The technique consists of 4 steps: pre-amplification, emulsion PCR, hybridization and flow cytometry. The determination of tissue KRAS profiling was performed by allele-specific RT-PCR (Cobas KRAS mutation test). Plasma samples from 23 patients were used for cell-free DNA (cfDNA) purification and cfDNA was further analyzed for 34 mutations in the KRAS and NRAS genes(exons 2,3,4) by BEAMing digital PCR. In 13 patients mutation was detected in exon 2 codon12 of KRAS gene (KR2-12), in 2 patients mutation was detected in KR2_12 and additionally, in exon 3 codon 61 of the NRAS gene (NR3_61) in one of them and in exon 2 codon 12 of NRAS in another patient. Tissue and plasma samples were analyzed for KRAS mutation for 11 patients. The concordance between plasma and tissue-based detection of KRAS mutations was high (9 out of 11 patients). In addition, 32 plasma samples from 16 patients have been analyzed for ctDNA RAS mutations. MAF values vary between 0,001-27,932% (MAF=RAS mutant alleles/RAS mutant+RAS wild-type alleles). Patient RAS Mutant allele fraction value (%) changed between the two time points (baseline and 4-6 weeks after). he number of mutant beads seems to have a prognostic role, as patients with a significantly high number of mutant beads tended to have a poor prognosis. At this point, MAF change has no significant correlation with CA19-9 change at the time points under study. This work highlighted the potential role of liquid biopsies in the management of PDAC and, more specifically, yielded preliminary data on the role of RAS ctDNA profiling as a predictive and prognostic marker. Kinetics of the RAS mutational load during therapy and its prognostic or predictive value will be analyzed with further cohort expansion." @default.
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• W3041299573 date "2020-07-01" @default.
• W3041299573 modified "2023-10-16" @default.
• W3041299573 title "P-181 Role of detection and quantification of plasma ctDNA RAS mutations by BEAMing digital PCR in patients with locally advanced and metastatic pancreatic cancer" @default.
• W3041299573 doi "https://doi.org/10.1016/j.annonc.2020.04.263" @default.
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