TY - JOUR

T1 - Multiplex Droplet Digital PCR Assay for Detection of MET and HER2 Genes Amplification in Non-Small Cell Lung Cancer

AU - Oscorbin, Igor P.

AU - Smertina, Maria A.

AU - Pronyaeva, Ksenia A.

AU - Voskoboev, Mikhail E.

AU - Boyarskikh, Ulyana A.

AU - Kechin, Andrey A.

AU - Demidova, Irina A.

AU - Filipenko, Maxim L.

N1 - Funding Information: Funding: This research was funded by Russian State funded budget project 121031300045-2 for IC-BFM SB RAS. The APC was funded by 121031300045-2. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2022/3/1

Y1 - 2022/3/1

N2 - Non-small-cell lung cancer (NSCLC), a subtype of lung cancer, remains one of the most common tumors with a high mortality and morbidity rate. Numerous targeted drugs were imple-mented or are now developed for the treatment of NSCLC. Two genes, HER2 and MET, are among targets for these specific therapeutic agents. Alterations in HER2 and MET could lead to primary or acquired resistance to commonly used anti-EGFR drugs. Using current methods for detecting HER2 and MET amplifications is time and labor-consuming; alternative methods are required for HER2 and MET testing. We developed the first multiplex droplet digital PCR assay for the simultaneous detection of MET and HER2 amplification in NSCLC samples. The suitability of qPCR was assessed for the optimization of multiplex ddPCR. The optimal elongation temperature, reference genes for DNA quantification, and amplicon length were selected. The developed ddPCR was validated on control samples with various DNA concentrations and ratios of MET and HER2 genes. Using ddPCR, 436 EGFR-negative NSCLC samples were analyzed. Among the tested samples, five specimens (1.15%) showed a higher ratio of MET, and six samples (1.38%) showed a higher ratio of HER2. The reported multiplex ddPCR assay could be used for the routine screening of MET and HER2 amplification in NSCLC samples.

AB - Non-small-cell lung cancer (NSCLC), a subtype of lung cancer, remains one of the most common tumors with a high mortality and morbidity rate. Numerous targeted drugs were imple-mented or are now developed for the treatment of NSCLC. Two genes, HER2 and MET, are among targets for these specific therapeutic agents. Alterations in HER2 and MET could lead to primary or acquired resistance to commonly used anti-EGFR drugs. Using current methods for detecting HER2 and MET amplifications is time and labor-consuming; alternative methods are required for HER2 and MET testing. We developed the first multiplex droplet digital PCR assay for the simultaneous detection of MET and HER2 amplification in NSCLC samples. The suitability of qPCR was assessed for the optimization of multiplex ddPCR. The optimal elongation temperature, reference genes for DNA quantification, and amplicon length were selected. The developed ddPCR was validated on control samples with various DNA concentrations and ratios of MET and HER2 genes. Using ddPCR, 436 EGFR-negative NSCLC samples were analyzed. Among the tested samples, five specimens (1.15%) showed a higher ratio of MET, and six samples (1.38%) showed a higher ratio of HER2. The reported multiplex ddPCR assay could be used for the routine screening of MET and HER2 amplification in NSCLC samples.

KW - Digital PCR

KW - Gene amplification

KW - HER2

KW - MET

KW - Multiplex ddPCR

KW - Non-small cell lung cancer

KW - NSCLC

KW - QPCR

KW - Real-time PCR

UR - http://www.scopus.com/inward/record.url?scp=85126303447&partnerID=8YFLogxK

U2 - 10.3390/cancers14061458

DO - 10.3390/cancers14061458

M3 - Article

C2 - 35326608

AN - SCOPUS:85126303447

VL - 14

JO - Cancers

JF - Cancers

SN - 2072-6694

IS - 6

M1 - 1458

ER -