Identification and functional analysis of acute myeloid leukemia susceptibility associated single nucleotide polymorphisms at non-protein coding regions ofRUNX1

Abstract

Little is known about the susceptibility to acute myeloid leukemia. We aim to search non-protein coding regions of key hematopoiesis transcription factors for genetic variations associated with acute myeloid leukemia susceptibility. We genotyped SNPs of RUNX1 P1 promoter, P2 promoter, +23 enhancer, intron 5.2 enhancer, PU.1 promoter, CEBPA promoter, and CEBPE promoter from acute myeloid leukemia patients and healthy controls. Rs2249650 and rs2268276 at RUNX1 intron 5.2 enhancer were found to be associated with acute myeloid leukemia susceptibility. Artificial reporters containing different rs2249650 and rs2268276 alleles showed differential activities in the K562 cell line, a human immortalized myeloid leukemia line. Rs2249650 contributes to reporter activities more than rs2268276. Gel shift assay is consistent with the luciferase assay. Supershift assay indicated that one potential binding protein was PU.1. To sum up, rs2268276 and especially rs2249650 may be qualified as new acute myeloid leukemia susceptibility-associated SNPs.

Transcriptional Regulation of RUNX1: An Informatics Analysis

“…Two SNPs in R1RE15 (rs933131 and rs2834716) were also found in patients with AML. Interestingly, the patient with R1RE15 rs2834716 also had the R1RE21 SNPs (rs2268276 and rs2249650) which were previously associated with AML susceptibility [92]. Five patients with malignant lymphoma had mutations within R1REs 1 and 10 that are not reported as SNPs with a minor allele frequency (MAF) of >1%.…”

“…GA has the least enhancer capability. The SNPs also affect SPI1 binding capability; AG has strong SPI1 binding, GA has weak SPI1 binding, whereas AA and GG have medium SPI1 binding capability [92].…”