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.

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.