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Highly Specific Droplet-Digital PCR Detection of Universally Methylated Circulating Tumor DNA in Endometrial Carcinoma.
Beinse, Guillaume; Borghese, Bruno; Métairie, Marie; Just, Pierre-Alexandre; Poulet, Geoffroy; Garinet, Simon; Parfait, Beatrice; Didelot, Audrey; Bourreau, Camille; Agueeff, Natacha; Lavollé, Alexandre; Terris, Benoit; Chapron, Charles; Goldwasser, François; Leroy, Karen; Blons, Helene; Laurent-Puig, Pierre; Taly, Valérie; Alexandre, Jérôme.
  • Beinse G; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Borghese B; Department of Medical Oncology, Hopital Cochin, Institut du Cancer Paris CARPEM, AP-HP, APHP Centre, Paris, France.
  • Métairie M; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Just PA; Department of Gynecological Surgery, Institut du Cancer Paris CARPEM, AP-HP, APHP Centre, Hopital Cochin, Paris, France.
  • Poulet G; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Garinet S; Department of Gynecological Surgery, Institut du Cancer Paris CARPEM, AP-HP, APHP Centre, Hopital Cochin, Paris, France.
  • Parfait B; Department of Pathology, Institut du Cancer Paris CARPEM, AP-HP, APHP Centre, Hopital Cochin, Paris, France.
  • Didelot A; Université de Paris, Paris, France.
  • Bourreau C; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Agueeff N; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Lavollé A; Department of Biology, Institut du Cancer Paris CARPEM, AP-HP, APHP Centre, Hopital Européen Georges Pompidou, Paris, France.
  • Terris B; Université de Paris, Paris, France.
  • Chapron C; Institut du Cancer Paris CARPEM, AP-HP, Fédération des Centres de Ressources Biologiques- Plateforme de Ressources Biologiques APHP, Centre de Ressources Biologiques-Site Cochin, Hôpital Cochin, Paris, France.
  • Goldwasser F; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Leroy K; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Blons H; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Laurent-Puig P; Centre de Recherche des Cordeliers, « Equipe labélisée Ligue Contre le Cancer ¼, CNRS SNC 5096, Sorbonne Université, Université de Paris, INSERM, Paris, France.
  • Taly V; Department of Pathology, Institut du Cancer Paris CARPEM, AP-HP, APHP Centre, Hopital Cochin, Paris, France.
  • Alexandre J; Université de Paris, Paris, France.
Clin Chem ; 68(6): 782-793, 2022 06 01.
Article en En | MEDLINE | ID: mdl-35323926
ABSTRACT

BACKGROUND:

No circulating biomarker is available for endometrial carcinoma (EC). We aimed to identify DNA positions universally hypermethylated in EC, and to develop a digital droplet PCR (ddPCR) assay for detection of hypermethylated circulating tumor DNA (meth-ctDNA) in plasma from patients with EC.

METHODS:

DNA positions hypermethylated in EC, and without unspecific hypermethylation in tissue/cell types releasing circulating cell-free DNA in plasma, were identified in silico from TCGA/Gene Expression Omnibus (GEO) data. A methylation-specific ddPCR (meth-ddPCR) assay following bisulfite conversion of DNA extracted from plasma was optimized for detection of meth-ctDNA according to dMIQE guidelines. Performances were validated on a retrospective cohort (n = 78 tumors, n = 30 tumor-adjacent tissues), a prospective pilot cohort (n = 33 stage I-IV patients), and 55 patients/donors without cancer.

RESULTS:

Hypermethylation of zinc finger and SCAN domain containing 12 (ZSCAN12) and/or oxytocin (OXT) classified EC samples from multiple noncancer samples with high diagnostic specificity/sensitivity [>97%; area under the curve (AUC) = 0.99; TCGA/GEO tissues/blood samples]. These results were confirmed in the independent retrospective cohort (AUC = 0.99). Meth-ddPCR showed a high analytical specificity (limit of blank = 2) and sensitivity (absolute lower threshold of detection = 50 pgmethDNA/mLplasma). In the pilot cohort, meth-ctDNA was detected in pretreatment plasma samples from 9/11 and 5/20 patients with advanced and non-advanced EC, respectively. 2 of 9 patients had ctDNA detected after macroscopic complete surgery and experienced progression within 6 months. No healthy donors had any copy of hypermethylated DNA detected in plasma.

CONCLUSIONS:

Meth-ddPCR of ZSCAN12/OXT allows a highly specific and sensitive detection of ctDNA in plasma from patients with EC and appears promising for personalized approaches for these patients.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Neoplasias Endometriales / ADN Tumoral Circulante Tipo de estudio: Diagnostic_studies / Guideline / Observational_studies / Prognostic_studies Límite: Female / Humans Idioma: En Año: 2022 Tipo del documento: Article
Texto completo
Imprimir
XML
PubMed Links
Search on Google

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Neoplasias Endometriales / ADN Tumoral Circulante Tipo de estudio: Diagnostic_studies / Guideline / Observational_studies / Prognostic_studies Límite: Female / Humans Idioma: En Año: 2022 Tipo del documento: Article