You are here

  • Home
  • Archive
  • Volume 64, Issue Suppl 1
  • PTH-171 Evaluation of TP53 mutations on barrett–s epithelium FFPE specimens with a novel next generation sequencing platform highlights a discordance with P53 immunohistochemistry: analysis of the barrett–s epithelium genome investigation study (begins)

Article Text

Article menu
Download PDFPDF
Oesophagus, Stomach, Duodenum III
PTH-171 Evaluation of TP53 mutations on barrett–s epithelium FFPE specimens with a novel next generation sequencing platform highlights a discordance with P53 immunohistochemistry: analysis of the barrett–s epithelium genome investigation study (begins)
  1. MA Butt1,
  2. S Khan2,
  3. R Haidry3,
  4. M Gandy4,
  5. I Puccio2,
  6. D Oukrif5,
  7. S Kerr3,
  8. E Bloom1,
  9. M Banks3,
  10. M Rodriguez-Justo5,
  11. M Novelli5,
  12. R Hamoudi2,
  13. L Lovat1
  1. 1Research Department of Tissue and Energy
  2. 2Integrated Molecular and Cell Science Laboratory, University College London
  3. 3Gastroenterology and Endoscopy, University College Hospital
  4. 4Pathology, The Doctors Laboratory
  5. 5Pathology, University College London, London, UK

Abstract

Introduction Genetic factors play a crucial role in the predisposition to Barrett’s epithelium (BE) and progression to oesophageal adenocarcinoma (OA). Using next generation sequencing (NGS) methods on frozen tissue, TP53 mutations have been identified in a stage specific manor in BE (1). The standard method of preserving BE is formalin fixing and paraffin embedding (FFPE). NGS on tiny FFPE specimens is challenging. We examined the prevalence of TP53 genomic aberrations and p53 expression in tiny FFPE BE biopsies using an adapted NGS platform.

Method 37 cases including non-dysplastic BE (NDBE; n = 7), low grade dysplasia (LGD; n = 7), high grade dysplasia (HGD; n = 9), intramucosal cancer (IMC; n = 6) and invasive OA (IOA; n = 8) were sequenced and immunostained. We used an inhouse, high throughput multiplexing method for targeted mutational screening using NGS on DNA extracted from FFPE tissue and analysed the files with our unique hidden Markov analytical method. Automated p53 IHC using clone D07 was used to correlate detected TP53 mutations with tissue expression.

Results DNA extracted varied from 2–50 ng, including some highly degraded DNA material. TP53 was mutated in 40.5% (15/37) of cases; 28.6% of NDBE (2/7), 71.4% of LGD (5/7), 44.4% of HGD (4/9), 33.3% of IMC (2/6) and 25% of IOA (2/8). Interestingly, only 40% (6/15) of cases showed both TP53 mutations and p53 aberrations (over- or absence of expression), 60% (9/15) had mutations but no protein aberration, and 45% (5/11) had p53 aberration but no mutational abnormality. The data quality was excellent with all cases having a depth of coverage of at least x800 (mean coverage x1200 (Figure 1A). The level of p53 staining did not however correlate with mutation data (Figure 1B).

Abstract PTH-171 Figure 1
Abstract PTH-171 Figure 1

Conclusion These results show discordance between detection of TP53 using mutation analysis and current p53 IHC methods. Furthermore, we identified patients with TP53 mutations in NDBE, which may pre-empt progression to OA. Identifying early genomic aberrations driving the development of OA from BE holds the key to accurately risk stratifying BE patients into more aggressive treatment strategies or frequent surveillance regimes.

Disclosure of interest None Declared.

Reference

  1. Weaver et al. Nat Genet. 2014;46(8):837-43

https://doi.org/10.1136/gutjnl-2015-309861.1059

Statistics from Altmetric.com

    Request Permissions

    If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.