title: A Study of Hypermethylated Circulating Tumor DNA as a Universal Colorectal Cancer Biomarker.
abstract: Circulating tumor DNA (ctDNA) has emerged as a good candidate for tracking tumor dynamics in different cancer types, potentially avoiding repeated tumor biopsies. Many different genes can be mutated within a tumor, complicating procedures for tumor monitoring, even with highly sensitive next-generation sequencing (NGS) strategies. Droplet-based digital PCR (dPCR) is a highly sensitive and quantitative procedure, allowing detection of very low amounts of circulating tumor genetic material, but can be limited in the total number of target loci monitored.
  
  
  
  We analyzed hypermethylation of 3 genes, by use of droplet-based dPCR in different stages of colorectal cancer (CRC), to identify universal markers for tumor follow-up.
  
  
  
  Hypermethylation of WIF1 (WNT inhibitory factor 1) and NPY (neuropeptide Y) genes was significantly higher in tumor tissue compared to normal tissue, independently of tumor stage. All tumor tissues appeared positive for one of the 2 markers. Methylated ctDNA (MetctDNA) was detected in 80% of metastatic CRC and 45% of localized CRC. For samples with detectable mutations in ctDNA, MetctDNA and mutant ctDNA (MutctDNA) fractions were correlated. During follow-up of different stage CRC patients, MetctDNA changes allowed monitoring of tumor evolution.
  
  
  
  These results indicate that MetctDNA could be used as a universal surrogate marker for tumor follow-up in CRC patients, and monitoring MetctDNA by droplet-based dPCR could avoid the need for monitoring mutations.
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Sonia
authors.initials: S
authors.lastname: Garrigou
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer; Department of Digestive Oncology, AP-HP, European Georges Pompidou Hospital, Paris Descartes University, Paris, France;
authors.forename: Geraldine
authors.initials: G
authors.lastname: Perkins
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Fanny
authors.initials: F
authors.lastname: Garlan
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Corinne
authors.initials: C
authors.lastname: Normand
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Audrey
authors.initials: A
authors.lastname: Didelot
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Delphine
authors.initials: D
authors.lastname: Le Corre
authors.affiliation: Department of Gastroenterology, Henri-Mondor Hospital-APHP and EA 7375-EC2M3 Laboratory, University of Paris Est Creteil Val de Marne, Creteil, France;
authors.forename: Sanam
authors.initials: S
authors.lastname: Peyvandi
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer; CRB Epigenetec, INSERM UMR-S1147, Centre Universitaire des Saints-Pères, Paris Cedex 06, France;
authors.forename: Claire
authors.initials: C
authors.lastname: Mulot
authors.affiliation: CIC-EC4 URC HEGP, AP-HP, Hôpitaux Universitaires Paris Ouest, Paris, France;
authors.forename: Ralph
authors.initials: R
authors.lastname: Niarra
authors.affiliation: CIC-EC4 URC HEGP, AP-HP, Hôpitaux Universitaires Paris Ouest, Paris, France;
authors.forename: Pascaline
authors.initials: P
authors.lastname: Aucouturier
authors.affiliation: CIC-EC4 URC HEGP, AP-HP, Hôpitaux Universitaires Paris Ouest, Paris, France;
authors.forename: Gilles
authors.initials: G
authors.lastname: Chatellier
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Philippe
authors.initials: P
authors.lastname: Nizard
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Karla
authors.initials: K
authors.lastname: Perez-Toralla
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer;
authors.forename: Eleonora
authors.initials: E
authors.lastname: Zonta
authors.affiliation: Department of Gastroenterology, Henri-Mondor Hospital-APHP and EA 7375-EC2M3 Laboratory, University of Paris Est Creteil Val de Marne, Creteil, France;
authors.forename: Cecile
authors.initials: C
authors.lastname: Charpy
authors.affiliation: INSERM U955, University of Paris Est Creteil Val de Marne and Department of Pathology, AP-HP, Henri-Mondor Hospital, Créteil, France;
authors.forename: Anais
authors.initials: A
authors.lastname: Pujals
authors.affiliation: CRB, AP-HP, Henri-Mondor Hospital, Créteil, France;
authors.forename: Caroline
authors.initials: C
authors.lastname: Barau
authors.affiliation: Service d'hépatogastroentérologie et de cancérologie digestive, CHU de Reims, Hôpital Robert-Debré, Reims Cedex, France;
authors.forename: Olivier
authors.initials: O
authors.lastname: Bouché
authors.affiliation: Department of Pathology, Hôpital Ambroise Paré, AP-HP, Université de Versailles St Quentin en Yvelines, Boulogne-Billancourt, France;
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Emile
authors.affiliation: CHU Clermont Ferrand, Clermont Ferrand Cedex 1, France;
authors.forename: Denis
authors.initials: D
authors.lastname: Pezet
authors.affiliation: Service d'Anatomo-Pathologie, Centre Val d'Aurelle Paul-Lamarque, Montpellier, France;
authors.forename: Frederic
authors.initials: F
authors.lastname: Bibeau
authors.affiliation: RainDance Technologies, Billerica, MA;
authors.forename: J Brian
authors.initials: JB
authors.lastname: Hutchison
authors.affiliation: RainDance Technologies, Billerica, MA;
authors.forename: Darren R
authors.initials: DR
authors.lastname: Link
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer; Department of Digestive Oncology, AP-HP, European Georges Pompidou Hospital, Paris Descartes University, Paris, France;
authors.forename: Aziz
authors.initials: A
authors.lastname: Zaanan
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer; Department of Biology, European Georges Pompidou Hospital, AP-HP, Paris Descartes University, Paris, France. valerie.taly@parisdescartes.fr pierre.laurent-puig@parisdescartes.fr iradj.sobhani@hmn.aphp.fr.
authors.forename: Pierre
authors.initials: P
authors.lastname: Laurent-Puig
authors.affiliation: Department of Gastroenterology, Henri-Mondor Hospital-APHP and EA 7375-EC2M3 Laboratory, University of Paris Est Creteil Val de Marne, Creteil, France; valerie.taly@parisdescartes.fr pierre.laurent-puig@parisdescartes.fr iradj.sobhani@hmn.aphp.fr.
authors.forename: Iradj
authors.initials: I
authors.lastname: Sobhani
authors.affiliation: Université Paris Sorbonne Cité, INSERM UMR-S1147, CNRS SNC5014, Centre Universitaire des Saints-Pères, Paris Cedex 06, France. Equipe labélisée Ligue contre le cancer; valerie.taly@parisdescartes.fr pierre.laurent-puig@parisdescartes.fr iradj.sobhani@hmn.aphp.fr.
authors.forename: Valerie
authors.initials: V
authors.lastname: Taly
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D000368:Aged; D014408:Biomarkers, Tumor / Q000235:genetics; D015179:Colorectal Neoplasms / Q000235:genetics*; D019175:DNA Methylation / Q000235:genetics; D004273:DNA, Neoplasm / Q000097:blood* / Q000737:chemistry* / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D009478:Neuropeptide Y / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012097:Repressor Proteins / Q000235:genetics*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D009478:Neuropeptide Y; D012097:Repressor Proteins; C485569:WIF1 protein, human