Heterogeneity in the colorectal primary tumor and the synchronous resected liver metastases prior to and after treatment with an anti-EGFR monoclonal antibody

Adua,Daniela; Di Fabio,Francesca; Ercolani,Giorgio; Fiorentino,Michelangelo; Gruppioni,Elisa; Altimari,Annalisa; Rojas Limpe,Fabiola  Lorena; Normanno,Nicola; Pinna,Antonio  Daniele; Pinto,Carmine

doi:10.3892/mco.2017.1270

Heterogeneity in the colorectal primary tumor and the synchronous resected liver metastases prior to and after treatment with an anti-EGFR monoclonal antibody

Authors:
- Daniela Adua
- Francesca Di Fabio
- Giorgio Ercolani
- Michelangelo Fiorentino
- Elisa Gruppioni
- Annalisa Altimari
- Fabiola Lorena Rojas Limpe
- Nicola Normanno
- Antonio Daniele Pinna
- Carmine Pinto
View Affiliations

Affiliations: Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, I‑40126 Bologna, Italy, Medical Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, I‑40138 Bologna, Italy, Department of General Surgery and Transplantation, Sant'Orsola‑Malpighi Hospital, University of Bologna, I‑40138 Bologna, Italy, Pathology Unit, Addarii Institute, Sant'Orsola‑Malpighi Hospital, University of Bologna, I‑40138 Bologna, Italy, Cell Biology and Biotherapy Unit, Istituto Nazionale Tumori ‘Fondazione Giovanni Pascale’ IRCCS, I‑80131 Naples, Italy, Medical Oncology Unit, Clinical Cancer Centre, IRCCS‑Arcispedale S. Maria Nuova, I‑42123 Reggio Emilia, Italy
Published online on: May 23, 2017 https://doi.org/10.3892/mco.2017.1270
Pages: 113-120

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Molecular heterogeneity between primary tumors (PTs) and synchronous resected liver metastasis in colorectal cancer (CRC) has potential relevance in treatment strategies. Next‑generation sequencing (NGS) may be able to increase the chances of identifying multiple molecular driver alterations, calling for therapy. The aim of the present study was to evaluate mutations in PT and synchronous resected liver metastases for patients with Kirsten rat sarcoma 2 viral oncogene homolog (KRAS) exon 2 wild‑type metastatic (m)CRC who underwent chemotherapy (CT) featuring an anti‑epidermal growth factor receptor (EGFR) monoclonal antibody. Genomic analysis was performed on 54 lesions from 7 patients with mCRC. For each patient, a PT biopsy or a surgical specimen was obtained prior to CT, and the PT and all liver metastases resected following CT were analyzed. DNA libraries were generated using the Ion AmpliSeq Colon and Lung Cancer Panel, assessing the most frequent somatic mutations in 22 genes involved in colon tumorigenesis, and sequencing was performed on an Ion Personal Genome Machine system. A partial response was achieved in all the patients, with a median progression free survival time of 11 months (range, 3‑21 months). All the patients were subjected to surgical liver metastasis resection. The median overall survival time was 31 months (range, 4‑46 months). Molecular analysis of the genes correlated with the target therapy, suggesting significant intratumor heterogeneity, as revealed by the different mutational landscape of certain PTs and synchronous resected liver metastases following systemic therapy when compared with the PT prior to treatment. In particular, the loss and acquisition of mutations in KRAS, neuroblastoma RAS viral oncogene homolog (NRAS), tumor protein p53 (TP53), the p110α catalytic subunit of phosphoinositide 3‑kinase (PIK3CA), F‑box/WD repeat‑containing protein 7 (FBXW7) and phosphatase and tensin homolog (PTEN) were observed. In addition, one patient developed a mucinous pattern following systemic CT. Taken together, the results of the present study demonstrated that intratumor heterogeneity is likely to affect the response to therapy, and to drive acquired resistance to targeted agents. The preliminary data also suggest a potential role for NGS in the evaluation of biological drug resistance, affecting future sequential treatment strategies.

View Figures

View References

1	Arnold M, Sierra MS, Laversanne M, Soerjomataram I, Jemal A and Bray F: Global patterns and trends in colorectal cancer incidence and mortality. Gut. 66:683–691. 2017. View Article : Google Scholar : PubMed/NCBI
2	Prenen H, Tejpar S and Van Cutsem E: Impact of molecular markers on treatment selection in advanced colorectal cancer. Eur J Cancer. 45 suppl 1:S70–S78. 2009. View Article : Google Scholar
3	Bokemeyer C, van Cutsem E, Rougier P, Ciardiello F, Heeger S, Schlichting M, Celik I and Köhne CH: Addition of cetuximab to chemotherapy as first-line treatment for KRAS wild-type metastatic colorectal cancer: Pooled analysis of the CRYSTAL and OPUS randomised clinical trials. Eur J Cancer. 48:1466–1475. 2012. View Article : Google Scholar : PubMed/NCBI
4	Douillard JY, Oliner KS, Siena S, Tabernero J, Burkes R, Barugel M, Humblet Y, Bodoky G, Cunningham D, Jassem J, et al: Panitumumab-FOLFOX4 treatment and RAS mutations in colorectal cancer. N Engl J Med. 369:1023–1034. 2013. View Article : Google Scholar : PubMed/NCBI
5	Stintzing S, Jung A, Rosssius L and Heinemann V: Analysis of KRAS/NRAS and BRAF mutations in FIRE-3: A randomized phase III study of Folfiri plus cetuximab or bevacizumab as first-line treatment for wild-type (WT) KRAS (exon 2) metastatic colorectal cancer (mCRC) patients. Eur J Cancer. 49 suppl 3:Abstract 17. 2013.
6	Schwartzberg LS, Rivera F, Karthaus M, Fasola G, Canon JL, Yu H, Oliner KS and Go WY: Analysis of KRAS/NRAS mutations in PEAK: A randomized phase II study of FOLFOX6 plus panitumumab (pmab) or bevacizumab (bev) as first-line treatment (tx) for wild-type (WT) KRAS (exon 2) metastatic colorectal cancer (mCRC). J Clin Oncol. 13 suppl 3:abstr 3631. 2013.
7	de Roock W, De Vriendt V, Normanno N, Ciardiello F and Tejpar S: KRAS, BRAF, PIK3CA, and PTEN mutations: Implications for targeted therapies in metastatic colorectal cancer. Lancet Oncol. 12:594–603. 2011. View Article : Google Scholar : PubMed/NCBI
8	Gerlinger M, Rowan AJ, Horswell S, Larkin J, Endesfelder D, Gronroos E, Martinez P, Matthews N, Stewart A, Tarpey P, et al: Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med. 366:883–892. 2012. View Article : Google Scholar : PubMed/NCBI
9	Kopetz S, Overman MJ, Chen K, Lucio-Eterovic AK, Kee BK, Fogelman DR, Dasari A, Raghav KPS, Sanchez EV, Phillips J, et al: Mutation and copy number discordance in primary versus metastatic colorectal cancer (mCRC). J Clin Oncol. 32 Suppl: abstr 3509:5S2014.
10	Adua D, Altimari A, Gruppioni E, Ercolani G, Llimpe FLR, Fabio FD, Fiorentino M, Pinna AD, Pinto C; Medical Oncology Unit, ; S. Orsola-Malpighi Hospital, Bologna, Italy, ; et al: Molecular evaluation of primary tumor (PT) and synchronous liver metastasis in colorectal cancer (srLmCRC) patients after cetuximab-based chemotherapy. J Clin Oncol. 32 Suppl:abstr 3624. 5S2014.
11	Mao C, Wu XY, Yang ZY, Threapleton DE, Yuan JQ, Yu YY and Tang JL: Concordant analysis of KRAS BRAF, PIK3CA mutations, and PTEN expression between primary colorectal cancer and matched metastases. Sci Rep. 5:80652015. View Article : Google Scholar : PubMed/NCBI
12	Sartore-Bianchi A, Di Nicolantonio F, Nichelatti M, Molinari F, De Dosso S, Saletti P, Martini M, Cipani T, Marrapese G, Mazzucchelli L, et al: Multi-determinants analysis of molecular alterations for predicting clinical benefit to EGFR-targeted monoclonal antibodies in colorectal cancer. PLoS One. 4:e72872009. View Article : Google Scholar : PubMed/NCBI
13	Normanno N, Rachiglio AM, Lambiase M, Martinelli E, Fenizia F, Esposito C, Roma C, Troiani T, Rizzi D, Tatangelo F, et al: Heterogeneity of KRAS NRAS, BRAF and PIK3CA mutations in metastatic colorectal cancer and potential effects on therapy in the CAPRI GOIM trial. Ann Oncol. 26:1710–174. 2015. View Article : Google Scholar : PubMed/NCBI
14	Ciardiello F, Normanno N, Maiello E, Martinelli E, Troiani T, Pisconti S, Giuliani F, Barone C, Cartenì G, Rachiglio AM, et al: Clinical activity of FOLFIRI plus cetuximab according to extended gene mutation status by next-generation sequencing: Findings from the CAPRI-GOIM trial. Ann Oncol. 25:1756–1761. 2014. View Article : Google Scholar : PubMed/NCBI
15	Ciardiello F and Normanno N: HER2 signaling and resistance to the anti-EGFR monoclonal antibody cetuximab: A further step toward personalized medicine for patients with colorectal cancer. Cancer Discov. 1:472–474. 2011. View Article : Google Scholar : PubMed/NCBI
16	Misale S, Di Nicolantonio F, Sartore-Bianchi A, Siena S and Bardelli A: Resistance to anti-EGFR therapy in colorectal cancer: From heterogeneity to convergent evolution. Cancer Discov. 4:1269–1280. 2014. View Article : Google Scholar : PubMed/NCBI
17	Yang ZY, Wu XY, Huang YF, Di MY, Zheng DY, Chen JZ, Ding H, Mao C and Tang JL: Promising biomarkers for predicting the outcomes of patients with KRAS wild-type metastatic colorectal cancer treated with anti-epidermal growth factor receptor monoclonal antibodies: A systematic review with meta-analysis. Int J Cancer. 133:1914–1925. 2013. View Article : Google Scholar : PubMed/NCBI
18	Guinney J, Ferté C, Dry J, McEwen R, Manceau G, Kao KJ, Chang KM, Bendtsen C, Hudson K, Huang E, et al: Modeling RAS phenotype in colorectal cancer uncovers novel molecular traits of RAS dependency and improves prediction of response to targeted agents in patients. Clin Cancer Res. 20:265–272. 2014. View Article : Google Scholar : PubMed/NCBI
19	Montagut C, Dalmases A, Bellosillo B, Crespo M, Pairet S, Iglesias M, Salido M, Gallen M, Marsters S, Tsai SP, et al: Identification of a mutation in the extracellular domain of the epidermal growth factor receptor conferring cetuximab resistance in colorectal cancer. Nat Med. 18:221–223. 2012. View Article : Google Scholar : PubMed/NCBI
20	Arena S, Bellosillo B, Siravegna G, Martínez A, Cañadas I, Lazzari L, Ferruz N, Russo M, Misale S, González I, et al: Emergence of multiple EGFR extracellular mutations during cetuximab treatment in colorectal cancer. Clin Cancer Res. 21:2157–2166. 2015. View Article : Google Scholar : PubMed/NCBI
21	van Emburgh BO, Arena S, Siravegna G, Lazzari L, Crisafulli G, Corti G, Mussolin B, Baldi F, Buscarino M, Bartolini A, et al: Acquired RAS or EGFR mutations and duration of response to EGFR blockade in colorectal cancer. Nat Commun. 7:136652016. View Article : Google Scholar : PubMed/NCBI
22	Montagut C, Siravegna G and Bardelli A: Liquid biopsies to evaluate early therapeutic response in colorectal cancer. Ann Oncol. 26:1525–1527. 2015. View Article : Google Scholar : PubMed/NCBI