title: Validation of a flow cytometry-based detection of γ-H2AX, to measure DNA damage for clinical applications.
abstract: The nucleosomal histone protein H2AX is specifically phosphorylated (γ-H2AX) adjacent to DNA double-strand breaks (DSBs) and is used for quantifying DSBs. Many chemotherapies and ionizing radiation (IR) used in cancer treatment result in DSBs. Therefore, γ-H2AX has a significant potential as a biomarker in evaluating patient sensitivity and responsiveness to IR and chemotherapy.
  
  
  
  Here, we report a flow cytometry-based quantification of γ-H2AX (FCM-γ-H2AX assay) customized for clinical practice.
  
  
  
  We validated that our method is able to detect DNA damage in peripheral blood mononuclear cells (PBMCs) treated with DSB inducing agents. The method also detected the DNA repair deficiency in PBMCs treated with DNA repair inhibitors, as well as the deficiency in DNA repair signaling in PBMCs from two ataxia telangiectasia patients.
  
  
  
  The FCM-γ-H2AX assay has sufficient analytical sensitivity and precision to measure levels of DNA damage and DNA repair for clinical purposes. © 2016 International Clinical Cytometry Society.
authors.affiliation: Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
authors.forename: Pegah
authors.initials: P
authors.lastname: Johansson
authors.affiliation: Department of Pediatrics, Institute of Clinical Sciences, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Fasth
authors.affiliation: Department of Pediatrics, Hospital of Halland, Halmstad, Sweden.
authors.forename: Torben
authors.initials: T
authors.lastname: Ek
authors.affiliation: Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
authors.forename: Ola
authors.initials: O
authors.lastname: Hammarsten
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology; D001260:Ataxia Telangiectasia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002867:Chromones / Q000494:pharmacology; D004247:DNA / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects*; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects / Q000528:radiation effects*; D004260:DNA Repair / Q000187:drug effects / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D053281:Enediynes / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods / Q000592:standards*; D005720:Gamma Rays; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects; D009025:Morpholines / Q000494:pharmacology; D010766:Phosphorylation; D061251:Primary Cell Culture; D011753:Pyrones / Q000494:pharmacology
chemical_list: C495818:2-morpholin-4-yl-6-thianthren-1-yl-pyran-4-one; C499693:8-dibenzothiophen-4-yl-2-morpholin-4-yl-chromen-4-one; D000617:Aminoglycosides; D002867:Chromones; D053281:Enediynes; C468783:H2AX protein, human; D006657:Histones; D009025:Morpholines; D011753:Pyrones; C074528:calicheamicin T; D004247:DNA
keywords: DNA double-strand break (DSB); H2AX; ataxia telangiectasia (AT); drug sensitivity; flow cytometry (FCM); gamma-H2AX; intrinsic radiation sensitivity; ionizing radiation (IR)