Abstract
Mutational heterogeneity could explain different metastatic patterns among IIIA-N2 lung cancer and influence prognosis. The identification of subclonal mutations using deep sequencing to evaluate the degree of molecular heterogeneity may improve IIIA-N2 classification. The aim of this prospective study was to assess mutational and immunohistochemical characteristics in primary tumours and involved lymph nodes (LN) in operated patients. Four patients operated for primary lung carcinoma and unisite N2 mediastinal involvement were consecutively selected. Samples (tumour and paired LN) were analysed for PD1, PD-L1 and CD8 immunostaining. Somatic mutation testing was performed by deep targeted next generation sequencing (NGS), with the AmpliSeq™ Colon and Lung Cancer Panel (LifeTechnology). A total of 9 primary lung cancer samples and 10 LN stations were analysed. For each cancer, we found 2 mutations, with allelic ratios from 3% to 72%. Mutational patterns were heterogeneous for 2 primary tumours. In 3 cases, mutations observed in the primary tumour were not found in LN metastases (ALK, FGFR3, MET). Inversely, in 1 case, a KRAS mutation was found in LN but not in the primary tumour. All primary tumours were found PD-L1 positive while CD8+ T cells infiltrate varied. In the different examined LN samples, PD-L1 expression, CD8+ and PD1+ T cells infiltrate were not similar to the primary tumour. This preliminary prospective study shows the diversity of intra-tumour and LN mutations using routinely-used targeted NGS, concerning both mutated gene and allelic ratio. Further studies are needed to evaluate its prognostic impact.
Similar content being viewed by others
References
Asamura H, Chansky K, Crowley J et al (2015) The International Association for the Study of Lung Cancer lung cancer staging project: proposals for the revision of the N descriptors in the forthcoming 8th edition of the TNM classification for lung cancer. J Thorac Oncol 10:1675–1684
Decaluwé H, De Leyn P, Vansteenkiste J et al (2009) Surgical multimodality treatment for baseline resectable stage IIIA-N2 non-small cell lung cancer. Degree of mediastinal lymph node involvement and impact on survival. Eur J Cardiothorac Surg 36:433–439
Arame A, Mordant P, Riquet M (2014) Pneumonectomy for stage IIIA NSCLC: a chance, not a calamity. Ann Thorac Surg 97:382
Riquet M, Mordant P, Pricopi C et al (2014) A review of 250 ten-year survivors after pneumonectomy for non-small-cell lung cancer. Eur J Cardiothorac Surg 45:876–881
Legras A, Mordant P, Arame A et al (2014) Long-term survival of patients with pN2 lung cancer according to the pattern of lymphatic spread. Ann Thorac Surg 97:1156–1162
Zhang J, Fujimoto J, Zhang J et al (2014) Intratumor heterogeneity in localized lung adenocarcinomas delineated by multiregion sequencing. Science 346:256–259
De Bruin EC, McGranahan N, Mitter R et al (2014) Spatial and temporal diversity in genomic instability processes defines lung cancer evolution. Science 346:251–256
Um S-W, Joung J-G, Lee H et al (2016) Molecular evolution patterns in metastatic lymph nodes reflect the differential treatment response of advanced primary lung cancer. Cancer Res 76:6568–6576
Pécuchet N, Legras A, Laurent-Puig P, Blons H (2016) Lung cancer molecular testing, what role for next generation sequencing and circulating tumor DNA. Ann Pathol 36:80–93
Garon EB, Rizvi NA, Hui R et al (2015) Pembrolizumab for the treatment of non-small-cell lung cancer. N Engl J Med 372:2018–2028
Mountain CF, Dresler CM (1997) Regional lymph node classification for lung cancer staging. Chest 111:1718–1723
Riquet M, Bagan P, Le Pimpec Barthes F et al (2007) Completely resected non-small cell lung cancer: reconsidering prognostic value and significance of N2 metastases. Ann Thorac Surg 84:1818–1824
Kozower BD, Larner JM, Detterbeck FC, Jones DR (2013) Special treatment issues in non-small cell lung cancer: diagnosis and management of lung cancer, 3rd ed: American College of Chest Physicians evidence-based clinical practice guidelines. Chest 143:e369S–e399S
Robinson LA, Ruckdeschel JC, Wagner H et al (2007) Treatment of non-small cell lung cancer-stage IIIA: ACCP evidence-based clinical practice guidelines (2nd edition). Chest 132:243S–265S
Cancer bronchique non à petites cellules - Référentiel national de RCP - Ref (2017) RECOKBRNONPETCEL15 | Institut National Du Cancer. http://www.e-cancer.fr/Expertises-et-publications/Catalogue-des-publications/Cancer-bronchique-non-a-petites-cellules-Referentiel-national-de-RCP. Accessed 26 June 2017
Jamal-Hanjani M, Quezada SA, Larkin J, Swanton C (2015) Translational implications of tumor heterogeneity. Clin Cancer Res 21:1258–1266
Hiley C, de Bruin EC, McGranahan N, Swanton C (2014) Deciphering intratumor heterogeneity and temporal acquisition of driver events to refine precision medicine. Genome Biol 15:453
Baldia PH, Maurer A, Heide T et al (2016) Fibroblast growth factor receptor (FGFR) alterations in squamous differentiated bladder cancer: a putative therapeutic target for a small subgroup. Oncotarget 7:71429–71439
Shirole NH, Pal D, Kastenhuber ER et al (2016) TP53 exon-6 truncating mutations produce separation of function isoforms with pro-tumorigenic functions. elife 6:e25532
Benderra M-A, Aspeslagh S, Postel-Vinay S et al (2016) Acquired EGFR mutation as the potential resistance driver to Crizotinib in a MET-mutated tumor. J Thorac Oncol 11:e21–e23
Waqar SN, Cottrell CE, Morgensztern D (2015) MET mutation associated with responsiveness to crizotinib. J Thorac Oncol 10:e29–e31
Koh J, Go H, Keam B et al (2015) Clinicopathologic analysis of programmed cell death-1 and programmed cell death-ligand 1 and 2 expressions in pulmonary adenocarcinoma: comparison with histology and driver oncogenic alteration status. Mod Pathol 28:1154–1166
D’Incecco A, Andreozzi M, Ludovini V et al (2015) PD-1 and PD-L1 expression in molecularly selected non-small-cell lung cancer patients. Br J Cancer 112:95–102
Vieira T, Antoine M, Hamard C et al (2016) Sarcomatoid lung carcinomas show high levels of programmed death ligand-1 (PD-L1) and strong immune-cell infiltration by TCD3 cells and macrophages. Lung Cancer Amst Neth 98:51–58
Uruga H, Bozkurtlar E, Huynh TG et al (2016) Programmed cell death ligand (PD-L1) expression in stage II and III lung adenocarcinomas and nodal metastases. J Thorac Oncol 12:458–466
Kim M-Y, Koh J, Kim S et al (2015) Clinicopathological analysis of PD-L1 and PD-L2 expression in pulmonary squamous cell carcinoma: comparison with tumor-infiltrating T cells and the status of oncogenic drivers. Lung Cancer Amst Neth 88:24–33
Taube JM, Klein A, Brahmer JR et al (2014) Association of PD-1, PD-1 ligands, and other features of the tumor immune microenvironment with response to anti-PD-1 therapy. Clin Cancer Res 20:5064–5074
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare that they have no conflict of interest.
Additional information
Hélène Blons and Françoise Le Pimpec-Barthes share senior authorship.
Rights and permissions
About this article
Cite this article
Legras, A., Roussel, H., Mangiameli, G. et al. Mutational Diversity of Lung Cancer and Associated Lymph Nodes. An Exploratory Prospective Study of 4 Resected cIIIA-N2. Pathol. Oncol. Res. 25, 319–325 (2019). https://doi.org/10.1007/s12253-017-0352-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12253-017-0352-x