Liquid biopsy has the potential to monitor biological effects of treatment. KRAS represents the most commonly mutated oncogene in Caucasian non-small-cell lung cancer (NSCLC). The aim of this study was to explore association of dynamic plasma KRAS genotyping with outcome in advanced NSCLC patients.
Advanced NSCLC patients were prospectively enrolled. Plasma samples were collected at baseline (T1), after 3 or 4 weeks, according to treatment schedule (T2) and at first radiological restaging (T3). Patients carrying KRAS mutation in tissue were analysed in plasma with droplet digital PCR. Semi-quantitative index of fractional abundance of mutated allele (MAFA) was used.
KRAS-mutated cohort included 58 patients, and overall 73 treatments (N = 39 chemotherapy and N = 34 immune checkpoint inhibitors) were followed with longitudinal liquid biopsy. Sensitivity of KRAS detection in plasma at baseline was 48.3% (95% confidence interval (CI): 35.0–61.8). KRAS mutation at T2 was associated with increased probability of experiencing progressive disease as best radiological response (adjusted odds ratio: 7.3; 95% CI: 2.1–25.0, p = 0.0016). Increased MAFA (T1–T2) predicted shorter progression-free survival (adjusted hazard ratio (HR): 2.1; 95% CI: 1.2–3.8, p = 0.0142) and overall survival (adjusted HR: 3.2; 95% CI: 1.2–8.4, p = 0.0168).
Longitudinal analysis of plasma KRAS mutations correlated with outcome: its early assessment during treatment has great potentialities for monitoring treatment outcome in NSCLC patients.
Subscribe to Journal
Get full journal access for 1 year
only $20.79 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Novello, S., Barlesi, F., Califano, R., Cufer, T., Ekman, S., Levra, M. G. et al. Metastatic non-small-cell lung cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann. Oncol. 27(Suppl. 5), v1–v27 (2016).
Rolfo, C., Mack, P. C., Scagliotti, G. V., Baas, P., Barlesi, F., Bivona, T. G. et al. Liquid biopsy for advanced non-small cell lung cancer (NSCLC): a statement paper from the IASLC. J. Thorac. Oncol. 13, 1248–1268 (2018).
Ettinger, D. S., Wood, D. E., Aggarwal, C., Aisner, D. L., Akerley, W., Bauman, J. R. et al. NCCN Guidelines Insights: non-small cell lung cancer, version 1.2020. J. Natl Compr. Cancer Netw. 17, 1464–1472 (2019).
Siravegna, G., Marsoni, S., Siena, S. & Bardelli, A. Integrating liquid biopsies into the management of cancer. Nat. Rev. Clin. Oncol. 14, 531–548 (2017).
Alix-Panabieres, C. & Pantel, K. Clinical applications of circulating tumor cells and circulating tumor DNA as liquid biopsy. Cancer Discov. 6, 479–491 (2016).
Diehl, F., Schmidt, K., Choti, M. A., Romans, K., Goodman, S., Li, M. et al. Circulating mutant DNA to assess tumor dynamics. Nat. Med. 14, 985–990 (2008).
Hu, Y., Ulrich, B. C., Supplee, J., Kuang, Y., Lizotte, P. H., Feeney, N. B. et al. False-positive plasma genotyping due to clonal hematopoiesis. Clin. Cancer Res. 24, 4437–4443 (2018).
Guibert, N., Mazieres, J., Delaunay, M., Casanova, A., Farella, M., Keller, L. et al. Monitoring of KRAS-mutated ctDNA to discriminate pseudo-progression from true progression during anti-PD-1 treatment of lung adenocarcinoma. Oncotarget 8, 38056–38060 (2017).
Yang, H., Liang, S. Q., Schmid, R. A. & Peng, R. W. New horizons in KRAS-mutant lung cancer: dawn after darkness. Front. Oncol. 9, 953 (2019).
Rex, K., Saiki, A. Y., Sun, J.-R., Holt, T., Koppada, N., Lanman, B. A. et al. Abstract 3090: in vivo characterization of AMG 510—a potent and selective KRASG12C covalent small molecule inhibitor in preclinical KRASG12 cancer models. Cancer Res. https://doi.org/10.1158/1538-7445.AM2019-3090 (2019).
Cabel, L., Riva, F., Servois, V., Livartowski, A., Daniel, C., Rampanou, A. et al. Circulating tumor DNA changes for early monitoring of anti-PD1 immunotherapy: a proof-of-concept study. Ann. Oncol. 28, 1996–2001 (2017).
Chaudhuri, A. A., Chabon, J. J., Lovejoy, A. F., Newman, A. M., Stehr, H., Azad, T. D. et al. Early detection of molecular residual disease in localized lung cancer by circulating tumor DNA profiling. Cancer Discov. 7, 1394–1403 (2017).
Kruger, S., Heinemann, V., Ross, C., Diehl, F., Nagel, D., Ormanns, S. et al. Repeated mutKRAS ctDNA measurements represent a novel and promising tool for early response prediction and therapy monitoring in advanced pancreatic cancer. Ann. Oncol. 29, 2348–2355 (2018).
Prentice, R. L., Williams, B. J. & Peterson, A. V. On the regression analysis of multivariate failure time data. Biometrika 68, 373–379 (1981).
Champiat, S., Dercle, L., Ammari, S., Massard, C., Hollebecque, A., Postel-Vinay, S. et al. Hyperprogressive disease is a new pattern of progression in cancer patients treated by anti-PD-1/PD-L1. Clin. Cancer Res. 23, 1920–1928 (2017).
Ferrara, R., Mezquita, L., Texier, M., Lahmar, J., Audigier-Valette, C., Tessonnier, L. et al. Hyperprogressive disease in patients with advanced non-small cell lung cancer treated with PD-1/PD-L1 inhibitors or with single-agent chemotherapy. JAMA Oncol. https://doi.org/10.1001/jamaoncol.2018.3676 (2018).
Sacher, A. G., Paweletz, C., Dahlberg, S. E., Alden, R. S., O'Connell, A., Feeney, N. et al. Prospective validation of rapid plasma genotyping for the detection of EGFR and KRAS mutations in advanced lung cancer. JAMA Oncol. 2, 1014–1022 (2016).
Reck, M., Hagiwara, K., Han, B., Tjulandin, S., Grohe, C., Yokoi, T. et al. ctDNA determination of EGFR mutation status in European and Japanese patients with advanced NSCLC: the ASSESS Study. J. Thorac. Oncol. 11, 1682–1689 (2016).
Doebele, R. C., Pilling, A. B., Aisner, D. L., Kutateladze, T. G., Le, A. T., Weickhardt, A. J. et al. Mechanisms of resistance to crizotinib in patients with ALK gene rearranged non-small cell lung cancer. Clin. Cancer Res. 18, 1472–1482 (2012).
Bordi, P., Tiseo, M., Rofi, E., Petrini, I., Restante, G., Danesi, R. et al. Detection of ALK and KRAS mutations in circulating tumor DNA of patients with advanced ALK-positive NSCLC with disease progression during crizotinib treatment. Clin. Lung Cancer 18, 692–697 (2017).
Garzon, M., Villatoro, S., Teixido, C., Mayo, C., Martinez, A., de Los Llanos Gil, M. et al. KRAS mutations in the circulating free DNA (cfDNA) of non-small cell lung cancer (NSCLC) patients. Transl. Lung Cancer Res. 5, 511–516 (2016).
Ai, B., Liu, H., Huang, Y. & Peng, P. Circulating cell-free DNA as a prognostic and predictive biomarker in non-small cell lung cancer. Oncotarget 7, 44583–44595 (2016).
Karachaliou, N., Mayo-de las Casas, C., Queralt, C., de Aguirre, I., Melloni, B., Cardenal, F. et al. Association of EGFR L858R mutation in circulating free DNA with survival in the EURTAC Trial. JAMA Oncol. 1, 149–157 (2015).
Marchetti, A., Palma, J. F., Felicioni, L., De Pas, T. M., Chiari, R., Del Grammastro, M. et al. Early prediction of response to tyrosine kinase inhibitors by quantification of EGFR mutations in plasma of NSCLC patients. J. Thorac. Oncol. 10, 1437–1443 (2015).
Yanagita, M., Redig, A. J., Paweletz, C. P., Dahlberg, S. E., O'Connell, A., Feeney, N. et al. A prospective evaluation of circulating tumor cells and cell-free DNA in EGFR-mutant non-small cell lung cancer patients treated with erlotinib on a phase II trial. Clin. Cancer Res. 22, 6010–6020 (2016).
Mok, T., Wu, Y. L., Lee, J. S., Yu, C. J., Sriuranpong, V., Sandoval-Tan, J. et al. Detection and dynamic changes of EGFR mutations from circulating tumor DNA as a predictor of survival outcomes in NSCLC patients treated with first-line intercalated erlotinib and chemotherapy. Clin. Cancer Res. 21, 3196–3203 (2015).
Goto, K., Ichinose, Y., Ohe, Y., Yamamoto, N., Negoro, S., Nishio, K. et al. Epidermal growth factor receptor mutation status in circulating free DNA in serum: from IPASS, a phase III study of gefitinib or carboplatin/paclitaxel in non-small cell lung cancer. J. Thorac. Oncol. 7, 115–121 (2012).
Goldberg, S. B., Narayan, A., Kole, A. J., Decker, R. H., Teysir, J., Carriero, N. J. et al. Early assessment of lung cancer immunotherapy response via circulating tumor DNA. Clin Cancer Res. 24, 1872–1880 (2018).
Abbosh, C., Birkbak, N. J., Wilson, G. A., Jamal-Hanjani, M., Constantin, T., Salari, R. et al. Phylogenetic ctDNA analysis depicts early-stage lung cancer evolution. Nature 545, 446–451 (2017).
Skoulidis, F., Goldberg, M. E., Greenawalt, D. M., Hellmann, M. D., Awad, M. M., Gainor, J. F. et al. STK11/LKB1 mutations and PD-1 inhibitor resistance in KRAS-mutant lung adenocarcinoma. Cancer Discov. https://doi.org/10.1158/2159-8290.CD-18-0099 (2018).
Bonanno, L., Zulato, E., Attili, I., Pavan, A., Del Bianco, P., Nardo, G. et al. Liquid biopsy as tool to monitor and predict clinical benefit from chemotherapy (CT) and immunotherapy (IT) in advanced non-small cell lung cancer (aNSCLC): a prospective study. Ann. Oncol. 29, viii649 (2018).
We thank Dr. Rafael Rosell for his critical review of the manuscript. This work was partially presented at the ESMO Congress in 2018.31
Ethics approval and consent to participate
The ethics committee of Istituto Oncologico Veneto evaluated and approved study design and informed consent (2016/82, 12 December 2016). Written informed consent was obtained from all patients before study entry. The study was conducted in accordance with the precepts of the Declaration of Helsinki.
Consent to publish
The property of data belongs to the sponsor, IOV, and the authors have the right to publish the data.
The data generated and analysed during this study are included in this published article and its additional files. Further raw data might be asked to the authors.
The authors declare no competing interests.
This work was funded by IOV intramural research grant 2017 – SINERGIA (to S.I. and L.B.). The QX200 ddPCR system (Bio-Rad Laboratories) was purchased through a grant provided by Università degli Studi di Padova, Padova, Italy (2015).
Note This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Zulato, E., Attili, I., Pavan, A. et al. Early assessment of KRAS mutation in cfDNA correlates with risk of progression and death in advanced non-small-cell lung cancer. Br J Cancer 123, 81–91 (2020). https://doi.org/10.1038/s41416-020-0833-7
Plasma-based longitudinal mutation monitoring as a potential predictor of disease progression in subjects with adenocarcinoma in advanced non-small cell lung cancer
BMC Cancer (2020)
Detection of KRAS G12/G13 Mutations in Cell Free-DNA by Droplet Digital PCR, Offers Prognostic Information for Patients with Advanced Non-Small Cell Lung Cancer
Clinically relevant prognostic and predictive markers for immune-checkpoint-inhibitor (ICI) therapy in non-small cell lung cancer (NSCLC)
BMC Cancer (2020)