Targeted next-generation sequencing for molecular diagnosis of endometriosis-associated ovarian cancer

J Mol Med (Berl). 2016 Jul;94(7):835-47. doi: 10.1007/s00109-016-1395-2. Epub 2016 Feb 27.

Abstract

Recent molecular and pathological studies suggest that endometriosis may serve as a precursor of ovarian cancer (endometriosis-associated ovarian cancer, EAOC), especially of the endometrioid and clear cell subtypes. Accordingly, this study had two cardinal aims: first, to obtain mutation profiles of EAOC from Taiwanese patients; and second, to determine whether somatic mutations present in EAOC can be detected in preneoplastic lesions. Formalin-fixed paraffin-embedded (FFPE) tissues were obtained from ten endometriosis patients with malignant transformation. Macrodissection was performed to separate four different types of cells from FFPE sections in six patients. The four types of samples included normal endometrium, ectopic endometriotic lesion, atypical endometriosis, and carcinoma. Ultra-deep (>1000×) targeted sequencing was performed on 409 cancer-related genes to identify pathogenic mutations associated with EAOC. The most frequently mutated genes were PIK3CA (6/10) and ARID1A (5/10). Other recurrently mutated genes included ETS1, MLH1, PRKDC (3/10 each), and AMER1, ARID2, BCL11A, CREBBP, ERBB2, EXT1, FANCD2, MSH6, NF1, NOTCH1, NUMA1, PDE4DIP, PPP2R1A, RNF213, and SYNE1 (2/10 each). Importantly, in five of the six patients, identical somatic mutations were detected in atypical endometriosis and tumor lesions. In two patients, genetic alterations were also detected in ectopic endometriotic lesions, indicating the presence of genetic alterations in preneoplastic lesion. Genetic analysis in preneoplastic lesions may help to identify high-risk patients at early stage of malignant transformation and also shed new light on fundamental aspects of the molecular pathogenesis of EAOC.

Key messages: Molecular characterization of endometriosis-associated ovarian cancer genes by targeted NGS. Candidate genes predictive of malignant transformation were identified. Chromatin remodeling, PI3K-AKT-mTOR, Notch signaling, and Wnt/β-catenin pathway may promote cell malignant transformation.

Keywords: Endometriosis; Malignant transformation; Mutation; Next-generation sequencing; Ovarian cancer.

MeSH terms

  • Adenocarcinoma, Clear Cell / diagnosis*
  • Adenocarcinoma, Clear Cell / genetics
  • Adenocarcinoma, Clear Cell / pathology
  • Adult
  • Aged
  • Carcinoma, Endometrioid / diagnosis*
  • Carcinoma, Endometrioid / genetics
  • Carcinoma, Endometrioid / pathology
  • Class I Phosphatidylinositol 3-Kinases / genetics*
  • Class I Phosphatidylinositol 3-Kinases / metabolism
  • DNA Mutational Analysis
  • DNA-Binding Proteins
  • Endometriosis / diagnosis*
  • Endometriosis / genetics
  • Endometriosis / pathology
  • Female
  • Gene Expression
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Middle Aged
  • Mutation
  • Neoplasm Staging
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / metabolism
  • Ovarian Neoplasms / diagnosis*
  • Ovarian Neoplasms / genetics
  • Ovarian Neoplasms / pathology
  • Paraffin Embedding
  • Receptors, Notch / genetics
  • Receptors, Notch / metabolism
  • Signal Transduction
  • Tissue Fixation
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Wnt Proteins / genetics
  • Wnt Proteins / metabolism

Substances

  • ARID1A protein, human
  • DNA-Binding Proteins
  • Nuclear Proteins
  • Receptors, Notch
  • Transcription Factors
  • Wnt Proteins
  • Class I Phosphatidylinositol 3-Kinases
  • PIK3CA protein, human