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77 Cervical pre-cancer vs invasive cancer: molecular differentiation with potential of improving cervical cancer screening worldwide
  1. C Banila1,
  2. B Nedjai1,
  3. C Reuter1,
  4. K Cuschieri2,
  5. G Clifford3 and
  6. A Lorincz1
  1. 1Queen Mary University of London, Wolfson Institute of preventive Medicine, London, UK
  2. 2University of Edinburgh, Royal Infirmary of Edinburgh, Edinburgh, UK
  3. 3World Health Organization, Institute for Research on Cancer, Lyon, France


Objectives The S5 DNA-methylation classifier, based on target CpG sites of the human gene EPB41L3, and viral late gene regions of HPV-16,18,31 and 33 (Lorincz A et al., 2016) has demonstrated better performance for detection of CIN2/3-women than either HPV16/18 genotyping, cytology or combination. We tested the performance of S5 in detecting invasive cancers versus pre-cancers and quantified the degree of separation between normal/CIN1, CIN2/3 and invasive cancer S5 scores.

Methods Methylation status of the S5-CpGs was tested in DNA extracted from exfoliated cervical cell from the UK(n=138),Spain(n=100),Colombia(n=96),Philippines(n=50),Georgia(n=42) and Ethiopia(n=79). Samples were histologically defined as negative/CIN1, CIN2/3 and invasive cancer. DNA-bisulfite conversion was carried out and followed by pyrosequencing for the 6 components of S5. Average methylation was calculated for each marker to define the S5 score.

Results Methylation at all sites increased proportionally with disease severity showing a Cuzick-trend of z=9.2933(p<2.2x10–16). The separation of normal/CIN1 from CIN2/3 and from cancer was highly-significant (Mann-Whitney, all p<0.0001). S5 also showed highly-significant difference between CIN2/3 and invasive cancer from matched cohorts: UK(p<0.003), Spain(p<0.0001) and Colombia(p<0.003). ROC-curves were used to assess the diagnostic potential of S5 in differentiating cancers from CIN2/3. The AUC was 0.86(CI 95%:0.7965 to 0.9131,p<0.0001) with a sensitivity of 79.8% and a specificity of 83.1%, based on a cut-off at highest Youden J-index.

Conclusions The S5 methylation classifier may be useful in cervical screening programs for identifying progressive pre-cancers in women. Although the separation was very good, there is room for improvement by addition of new markers derived from our ongoing NGS multi-omics study.

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