title: The rate of spontaneous mutations in human myeloid cells.
abstract: The mutation rate (μ) is likely to be a key parameter in leukemogenesis, but historically, it has been difficult to measure in humans. The PIG-A gene has some advantages for the detection of spontaneous mutations because it is X-linked, and therefore only one mutation is required to disrupt its function. Furthermore, the PIG-A-null phenotype is readily detected by flow cytometry. Using PIG-A, we have now provided the first in vitro measurement of μ in myeloid cells, using cultures of CD34+ cells that are transduced with either the AML-ETO or the MLL-AF9 fusion genes and expanded with cytokines. For the AML-ETO cultures, the median μ value was ∼9.4×10(-7) (range ∼3.6-23×10(-7)) per cell division. In contrast, few spontaneous mutations were observed in the MLL-AF9 cultures. Knockdown of p53 or introduction of mutant NRAS or FLT3 alleles did not have much of an effect on μ. Based on these data, we provide a model to predict whether hypermutability must occur in the process of leukemogenesis.
authors.affiliation: Division of Hematology, Department of Veterans Affairs New York Harbor Healthcare System, United States; Division of Hematology, Department of Medicine, NYU School of Medicine and the NYU Langone Cancer Center, United States. Electronic address: david.araten@nyumc.org.
authors.forename: David J
authors.initials: DJ
authors.lastname: Araten
authors.forename: Ondrej
authors.initials: O
authors.lastname: Krejci
authors.forename: Kimberly
authors.initials: K
authors.lastname: Ditata
authors.forename: Mark
authors.initials: M
authors.lastname: Wunderlich
authors.forename: Katie J
authors.initials: KJ
authors.lastname: Sanders
authors.forename: Leah
authors.initials: L
authors.lastname: Zamechek
authors.forename: James C
authors.initials: JC
authors.lastname: Mulloy
mesh_terms: D002471:Cell Transformation, Neoplastic / Q000235:genetics; D002478:Cells, Cultured; D050676:Core Binding Factor Alpha 2 Subunit / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques; D016158:Genes, p53; D017261:Glycosylphosphatidylinositols / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000502:physiology; D059645:Mutation Rate; D022423:Myeloid Cells / Q000166:cytology / Q000378:metabolism*; D051788:Myeloid-Lymphoid Leukemia Protein / Q000502:physiology; D015514:Oncogene Proteins, Fusion / Q000502:physiology; D000075142:RUNX1 Translocation Partner 1 Protein
chemical_list: C107844:AML1-ETO fusion protein, human; D050676:Core Binding Factor Alpha 2 Subunit; D017261:Glycosylphosphatidylinositols; C474558:MLL-AF9 fusion protein, human; D008565:Membrane Proteins; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein; C080086:phosphatidylinositol glycan-class A protein; D051788:Myeloid-Lymphoid Leukemia Protein
keywords: AML; B-lymphoblastoid cell lines; BLCLs; GPI; GPI-linked proteins; Human myeloid cultures; Mutation rate; Myeloid leukemia.; PIG-A gene; PNH; Spontaneous somatic mutations; acute myelogenous leukemia; glycosylphosphatidylinositol; paroxysmal nocturnal hemoglobinuria