Abstract 1912: ADAR1-mediated microRNA regulation and blast crisis leukemia stem cell generation in chronic myeloid leukemia
Authors: Zipeto Maria , Jiang Qingfei , Robertson Leslie Crews , Jamieson Catriona HM 
Published: May 2015

Introduction

Chronic myeloid leukemia (CML) was one of the first malignancies shown to be initiated in hematopoietic stem cells by the BCR-ABL1 oncogene and sustained in blast crisis (BC) by progenitor cells that co-opt stem cell properties and behave like leukemia stem cells (BC LSC). The BCR-ABL fusion oncogene encodes a constitutively active tyrosine kinase BCR-ABL. Although tyrosine kinase inhibitor (TKI) therapy targeting BCR-ABL suppresses CML during the chronic phase (CP), progenitors undergo expansion as a consequence of subsequent genetic and epigenetic alterations that fuel blast crisis (BC) transformation, BC LSC generation and TKI resistance. We have recently shown that adenosine to insoine (A-to-I) RNA editing activity, mediated by the adenosine deaminase acting on dsRNAs ADAR1, is enhanced in BC CML and drives leukemic progression by inducing myeloid progenitor reprogramming and enhancing self-renewal. ADAR1 targets double-stranded RNA hairpin-containing loop structures, such as microRNAs (miRNAs). MiRNAs, by operating through base-pairing with complementary sequences within a mRNA molecule lead to mRNA degradation and gene silencing. Emerging evidence suggests miRNAs alterations introduced at the pri-miRNA or pre-miRNA level by A-to-I editing by ADAR1 may be responsible for broad changes in the transcriptome that drives LSC maintenance. However, the overall change in edited miRNA profiles in CP CML compared with BC progenitors is unclear. In this study we aimed to investigate the role of ADAR1 in driving malignant progression by focusing on its regulation of miRNA expression in CML.

Methods and Results

MicroRNA expression profiles were evaluated in CD34+ cells derived from CP and BC CML patient samples as well as cord blood. MiRNA PCR array (Qiagen) allowed us to identify differentially expressed miRNAs that were subsequently validated by q-PCR with specific primers targeting individual miRNA. We observed the downregulation of 13 miRNA in CML BC compared to CML CP progenitors. Lentiviral overexpression of ADAR1 in CD34+ cord blood cells led to significant downregulation of 17 miRNAs, including members of the let-7 family, and other miRNAs involved in the regulation of self-renewal. Interestingly, BC CD34+ cells showed a miRNA profile similar to that of lentiviral ADAR1 transduced cord blood progenitors. Lentiviral overexpression of ADAR1 in CD34+ chronic phase CML cells, led to a significant downregulation of 14 miRNAs compared with their lentiviral backbone transduced counterparts.

Conclusion

Our data suggest that ADAR1 contributes to BC LSC generation by downregulating miRNAs that target stem cell regulatory gene products. A further characterization of ADAR-editing related miRNA signatures may enable BC LSC detection and early intervention with combined BCR-ABL1 and ADAR1-targeting strategies that may obviate CML relapse and progression.

Citation Format: Maria Zipeto, Qingfei Jiang, Leslie Crews Robertson, Catriona HM Jamieson. ADAR1-mediated microRNA regulation and blast crisis leukemia stem cell generation in chronic myeloid leukemia. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1912. doi:10.1158/1538-7445.AM2014-1912