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DNA hypermethylation analysis in sputum of asymptomatic subjects at risk for lung cancer participating in the NELSON trial: argument for maximum screening interval of 2 years
  1. A Jasmijn Hubers1,
  2. Daniëlle A M Heideman1,
  3. Sylvia Duin1,
  4. Birgit I Witte2,
  5. Harry J de Koning3,
  6. Harry J M Groen4,
  7. Clemens F M Prinsen5,
  8. Anne S Bolijn1,5,
  9. Mandy Wouters5,
  10. Susanne E van der Meer5,
  11. Renske D M Steenbergen1,
  12. Peter J F Snijders1,
  13. Anne Uyterlinde1,
  14. Hans Berkhof2,
  15. Egbert F Smit6,
  16. Erik Thunnissen1
  1. 1Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands
  2. 2Department of Epidemiology and Biostatistics, VU University Medical Center, Amsterdam, The Netherlands
  3. 3Department of Public Health, Erasmus Medical Center, Rotterdam, The Netherlands
  4. 4Department of Pulmonary Diseases, University Medical Center Groningen, Groningen, The Netherlands
  5. 5Department of Pathology, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands
  6. 6Department of Pulmonary Diseases, VU University Medical Center, Amsterdam, The Netherlands
  1. Correspondence to Dr Erik Thunnissen, Department of Pathology, VU University Medical Center, De Boelelaan 1117, Amsterdam 1081 HV, The Netherlands; e.thunnissen{at}


Aims Lung cancer is the major contributor to cancer mortality due to metastasised disease at time of presentation. The current study investigated DNA hypermethylation of biomarkers RASSF1A, APC, cytoglobin, 3OST2, FAM19A4, PHACTR3 and PRDM14 in sputum of asymptomatic high-risk individuals from the NELSON lung cancer low-dose spiral CT screening trial to detect lung cancer at preclinical stage.

Methods Subjects were selected with (i) lung cancer in follow-up (cases; n=65), (ii) minor cytological aberrations (controls; n=120) and (iii) a random selection of subjects without cytological aberrations (controls; n=99). Median follow-up time for controls was 80 months. Cut-off values were based on high specificity to assess diagnostic value of the biomarkers.

Results RASSF1A may denote presence of invasive cancer because of its high specificity (93% (95% CI 89% to 96%); sensitivity 17% (95% CI 4% to 31%), with best performance in a screening interval of 2 years. The panel of RASSF1A, 3OST2 and PRDM14 detected 28% (95% CI 11% to 44%) of lung cancer cases within 2 years, with specificity of 90% (95% CI 86% to 94%). Sputum cytology did not detect any lung cancers.

Conclusions In a lung cancer screening setting with maximum screening interval of 2 years, DNA hypermethylation analysis in sputum may play a role in the detection of preclinical disease, but complementary diagnostic markers are needed to improve sensitivity.

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  • Handling editor Runjan Chetty

  • Funding This study was funded by Dutch Cancer Society (grant VU2008-4220) and Perceptronix Medical (Vancouver, Canada).

  • Competing interests HJdK reports grants from ZonMw, Dutch Cancer Society, RVVZ, Roche Diagnostics, other from Siemens Germany and LungCARE, during the conduct of the study. ET reports grants for sputum examination from Dutch Cancer Society, MdxHealth.

  • Ethics approval Dutch Ministry of Health (NL 22206.029.08) and the Institutional Review Boards.

  • Provenance and peer review Not commissioned; externally peer reviewed.