Skip to main content

Advertisement

SpringerLink
Log in

Targeted deep sequencing helps distinguish independent primary tumors from intrapulmonary metastasis for lung cancer diagnosis

  • Original Article – Clinical Oncology
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

Multiple lung lesions found in a single patient at the time of diagnosis often pose a diagnostic dilemma: are these lesions independent primary tumors (IPT) or the result of intrapulmonary metastases (IPM)? While traditional pathological methods sometimes have difficulty distinguishing IPM from IPT, modern molecular profiling based on next-generation sequencing techniques may provide a new strategy.

Methods

Sixteen patients with multiple tumors were enrolled in this study. We performed targeted deep sequencing (~ 2000 × coverage) on a total of 40 tumors and matched blood samples. We compared mutational profiles between tumors within each patient and across patients to evaluate if they were genetically related. Computed tomographic images and histological staining were also used to validate tumor relationships.

Results

A total of 125 mutations were identified in 16 patients. Twelve out of fourteen patients whose histological diagnoses favored IPT did not have any shared mutations in their multiple tumors. The other two showed discrepancies: Pt01 had a shared EGFR exon19 deletion in the two lung tumors found, and Pt16 had one common mutation (BRAFD594G) in two out of five lung tumors. Pt14 with lung metastasis from salivary gland adenoid cystic carcinoma had shared mutations; and Pt15 with suspected intrapulmonary metastasis (IPM) had identical mutations between the two tumors. Visualized data can be readily accessed through the website: mlc.opengene.org.

Conclusion

Analysis of overlapping mutations among different tumors assists physicians in distinguishing IPM from IPT. Our findings demonstrate that DNA sequencing can provide additional evidence in clinical practice when pathology is inadequate to make a conclusive diagnosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Availability of data and materials

To access the supplementary material accompanying this article, visit the online version of the journal at https://www.springer.com/journal/432/. Raw datasets analyzed during the current study are available from the corresponding author upon reasonable request.

Abbreviations

IPT:

Independent primary tumor

IPM:

Intrapulmonary metastasis

NGS:

Next generation sequencing

CT:

Computed tomography

ctDNA:

Circulating tumor DNA

VAF:

Variant allele frequency

RUL:

Right upper lobe

LUL:

Left upper lobe

AIS:

Adenocarcinoma in situ

MIA:

Minimally invasive adenocarcinoma

ACC:

Adenoid cystic carcinoma

References

  • Aokage K, Ishii G, Nagai K et al (2007) Intrapulmonary metastasis in resected pathologic stage IIIB non-small cell lung cancer: possible contribution of aerogenous metastasis to the favorable outcome. J Thorac Cardiovasc Surg 134(2):386–391

    Article  Google Scholar 

  • Chen K, Chen W, Cai J et al (2018a) Favorable prognosis and high discrepancy of genetic features in surgical patients with multiple primary lung cancers. J Thorac Cardiovasc Surg 155(1):371–379

    Article  Google Scholar 

  • Chen S, Zhou Y, Chen Y et al (2019) Gencore: an efficient tool to generate consensus reads for error suppressing and duplicate removing of NGS data. BMC Bioinforma 20(Suppl 23):606

    Article  Google Scholar 

  • Chen S, Zhou Y, Chen Y, Gu J (2018c) Fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 34(17):i884–i890

    Article  Google Scholar 

  • Cheng DT, Mitchell TN, Zehir A et al (2015) Memorial sloan kettering-integrated mutation profiling of actionable cancer targets (MSK-IMPACT): a hybridization capture-based next-generation sequencing clinical assay for solid tumor molecular oncology. J Mol Diagn 17(3):251–264

    Article  CAS  Google Scholar 

  • Fabian T, Bryant AS, Mouhlas AL, Federico JA, Cerfolio RJ (2011) Survival after resection of synchronous non-small cell lung cancer. J Thorac Cardiovasc Surg 142(3):547–553

    Article  Google Scholar 

  • Gaikwad A, Souza CA, Inacio JR et al (2014) Aerogenous metastases: a potential game changer in the diagnosis and management of primary lung adenocarcinoma. Am J Roentgenol 203(6):W570–W582

    Article  Google Scholar 

  • Girard N, Pao W, Deshpande C et al (2009) Comprehensive histologic assessment helps to differentiate multiple lung primary nonsmall cell carcinomas from metastases. Am J Surg Pathol 33(12):1752–1764

    Article  Google Scholar 

  • Govindan R, Ding L, Griffith M et al (2012) Genomic landscape of non-small cell lung cancer in smokers and never-smokers. Cell 150:1121–1134

    Article  CAS  Google Scholar 

  • Koboldt DC, Zhang Q, Larson DE et al (2012) VarScan 2: somatic mutation and copy number alteration discovery in cancer by exome sequencing. Genome Res 22(3):568–576

    Article  CAS  Google Scholar 

  • Lawrence MS, Stojanov P, Polak P et al (2013) Mutational heterogeneity in cancer and the search for new cancer-associated genes. Nature 499:214

    Article  CAS  Google Scholar 

  • Li H, Durbin R (2010) Fast and accurate long-read alignment with Burrows–Wheeler transform. Bioinformatics 26(5):589–595

    Article  Google Scholar 

  • Li H, Handsaker B, Wysoker A et al (2009) The sequence alignment/map format and SAMtools. Bioinformatics 25(16):2078–2079

    Article  Google Scholar 

  • Li R, Li X, Xue R et al (2018) Early metastasis detected in patients with multifocal pulmonary ground-glass opacities (GGOs). Thorax 73(3):290–292

    Article  Google Scholar 

  • Liu Y, Zhang J, Li L et al (2016) Genomic heterogeneity of multiple synchronous lung cancer. Nat Commun. 7:1–8

    CAS  Google Scholar 

  • Macmahon H, Naidich DP, Goo JM et al (2017) Guidelines for management of incidental pulmonary nodules MacMahon et al. Radiol Radiol 284:228–243

    Article  Google Scholar 

  • Martini N, Melamed MR (1975) Multiple primary lung cancers. J Thorac Cardiovasc Surg 70(4):606–612

    Article  CAS  Google Scholar 

  • Murphy SJ, Aubry MC, Harris FR et al (2014) Identification of independent primary tumors and intrapulmonary metastases using DNA rearrangements in non-small-cell lung cancer. J Clin Oncol 32(36):4050–4058

    Article  CAS  Google Scholar 

  • Pomplun S (2006) Pathology of lung cancer. Lung Cancer 32(4):12–26

    Article  Google Scholar 

  • Sweeney SM, Cerami E, Baras A et al (2017) AACR project genie: powering precision medicine through an international consortium. Cancer Discov 7(8):818–831

    Article  Google Scholar 

  • Team TNLSTR (2011) Reduced lung-cancer mortality with low-dose computed tomographic screening. N Engl J Med. 365(5):395–409

    Article  Google Scholar 

  • Wu C, Zhao C, Yang Y et al (2015) High discrepancy of driver mutations in patients with NSCLC and synchronous multiple lung ground-glass nodules. J Thorac Oncol. 10(5):778–783

    Article  CAS  Google Scholar 

  • Yatabe Y, Kerr KM, Utomo A et al (2015) EGFR mutation testing practices within the Asia pacific region: results of a multicenter diagnostic survey. J Thorac Oncol 10(3):438–445

    Article  CAS  Google Scholar 

  • Zehir A, Benayed R, Shah RH et al (2017) Mutational landscape of metastatic cancer revealed from prospective clinical sequencing of 10,000 patients. Nat Med 23(6):703–713

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We acknowledge the generous gift of clinical specimens from our patients, to whom we dedicate our work. We thank Jinhua Zhong for clinical sample coordination.

Funding

We thank grants from the Natural Science Foundation of Guangdong Province of China (2017A030310641), the Medical Scientific Research Foundation of Guangdong Province of China (A2017327) for study design and data collection, the Science and Technology Innovation Committee of Shenzhen Municipality (JCYJ20180228175531145) for data collection and analysis, the Shenzhen Strategic Emerging Industry Development Special Fund (20170922151538732) and the PUHSC-UMHS Joint Institute Project (2019020(PUSH)-r1) for interpretation and writing.

Author information

Author notes

  1. Jixian Liu, Guangxian Mao, and Yingmei Li contributed equally to this work.

Authors and Affiliations

  1. Department of Thoracic Surgery, Peking University Shenzhen Hospital, 1120 Lianhua Road, Shenzhen, 518035, People’s Republic of China

    Jixian Liu, Guangxian Mao, Xuxing Peng, Junbin Wang, Xiaoqiang Li, Xinyu Luan, Ruixing Luo & Da Wu

  2. HaploX Biotechnology, 3 Songpingshan Road, Fl 8, Shenzhen, 518057, People’s Republic of China

    Yingmei Li, Weifeng Wang, Tanxiao Huang, Jing Zhang, Mengmeng Xu & Shifu Chen

  3. Department of Pathology, Peking University Shenzhen Hospital, Shenzhen, People’s Republic of China

    Lili Tao

  4. Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, People’s Republic of China

    Shifu Chen

Authors
  1. Jixian Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guangxian Mao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yingmei Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lili Tao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Weifeng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xuxing Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Junbin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiaoqiang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Xinyu Luan
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ruixing Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Tanxiao Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jing Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Mengmeng Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Shifu Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Da Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

JL, YL, SC and DW designed the study; LT performed the histological examination; JL, GM, XP, JW, XL and RL analyzed and interpreted the patient clinical data; MX and TH carried out the sequencing experiment and collected data; YL, WW, JZ and SC analyzed the bioinformatic data; YL, JL, XL and SC were major contributors in writing manuscript; SC and DW supervised the study.

Corresponding authors

Correspondence to Shifu Chen or Da Wu.

Ethics declarations

Conflict of interest

The authors declare no conflicts of interest.

Ethics approval and consent to participate

Clinical samples were obtained through the Peking University Shenzhen Hospital Institutional Review Board-approved informed consent process. No procedures were conducted for the exclusive purpose of research.

Consent for publication

Not applicable.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, J., Mao, G., Li, Y. et al. Targeted deep sequencing helps distinguish independent primary tumors from intrapulmonary metastasis for lung cancer diagnosis. J Cancer Res Clin Oncol 146, 2359–2367 (2020). https://doi.org/10.1007/s00432-020-03227-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00432-020-03227-5

Keywords

Search

Navigation