Transcriptome-Wide Analysis of RNA N6-Methyladenosine Modification in Adriamycin-Resistant Acute Myeloid Leukemia Cells

Abstract

Acute myeloid leukemia (AML) is one of the most aggressive hematopoietic malignancies. Patients still suffer from refractory/relapsed disease after anthracycline-based therapy, which leads to a poor prognosis. N6-Methyladenosine (m6A) is the most abundant post-transcriptional modification in eukaryotes, the imbalance of which is reported to be associated with various pathological processes, including drug resistance. However, the relationship between m6A modification and drug resistance has not been well defined in AML. In this study, we analyzed the sequencing data of HL60 and its Adriamycin-resistant cell line HL60/ADR. We found a total of 40,550 m6A-methylated peaks, representing 15,640 genes in HL60, and 38,834 m6A-methylated peaks, representing 15,285 genes in HL60/ADR. KEGG pathway analysis showed that pathways were enriched in the FoxO signaling pathway, p53 signaling pathway, and Notch signaling pathway. MeRIP-seq results showed that the fold enrichment of the global m6A level in HL60/ADR was higher than that in HL60, and dot blot assay results indicated that the global m6A level was elevated in HL60/ADR cells compared with that in HL60 cells. Further analysis revealed that the expression level of METTL3 was elevated in HL60/ADR cells compared with that in HL60 cells. After a combined treatment of STM2457 (an inhibitor of METTL3) and Adriamycin, the proliferation of HL60/ADR was inhibited. Thus, we hypothesized that the abnormality of m6A modification played an important role in Adriamycin-resistant AML.

Genome-wide investigation of differentially expressed alternative splicing and RNA-binding protein genes association with AML drug resistance

“…However, a majority of relapsed or drug-resistant AML patients have mutations in the deoxycytidine kinase gene, resulting in increased resistance to pharmacotherapy and eventual treatment failure [28]. Recently, studies have shown that mRNA methylation under the regulation of methyltransferase 3 (METTL3) may be involved in the regulation of drug resistance in AML [29,30]. Here, we report the signi cant alteration in global gene expression pro les in mononuclear cells from bone marrow blood in relapsed or drug-resistant AML patients compared with AML remission and Healthy controls.…”

Small molecule inhibitors targeting m6A regulators

“…While METTL3 and FTO have opposing roles in the regulation of m 6 A modification, it's noteworthy that due to differences in the signaling pathways involved, genetic knockdown of either of these genes enhances the sensitivity of BC cells to doxorubicin. METTL3 has also been found to be involved in doxorubicin resistance of AML cells, and inhibition of METTL3 increased the sensitivity of drug-resistant cells and inhibited the proliferation of doxorubicin resistant HL60/ADR cells [ 157 ]. Moreover, WTAP has also been implicated in the resistance of AML cells to doxorubicin [ 158 ].…”