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Targeting O-GlcNAcylated METTL3 impedes MDS/AML progression via diminishing SRSF1 m6A modification
Junjie Gou1, Yi Wang2, Jingjing Feng3
1Xi'an No. 1 Hospital, First Affiliated Hospital of Northwest University, School of Medicine, Xi'an, China; Key Laboratory of Resource Biology and Biotechnology of Western China, Ministry of Education, Provincial Key Laboratory of Biotechnology, College of Life Sciences, Northwest University, Xi'an, China.
Abstract:
N6-methyladenosine (m6A) modification, primarily regulated by methyltransferase-like protein 3 (METTL3), plays a pivotal role in RNA metabolism and leukemogenesis. However, the post-translational mechanisms governing METTL3 stability and function remain incompletely understood. Given the widespread occurrence of O-GlcNAcylation on nuclear and cytosolic proteins, we hypothesized that METTL3 might undergo O-GlcNAcylation, thereby influencing its stability and oncogenic function in myeloid malignancies. In this study, we found that METTL3 is O-GlcNAcylated in both myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML), and its expression positively correlates with O-GlcNAcylation levels. Functional assays demonstrated that O-GlcNAcylation enhances METTL3 protein stability and promotes leukemic cell survival. Mechanistically, O-GlcNAcylated METTL3 stabilizes mRNA of serine- and arginine-rich splicing factor 1 (SRSF1), leading to increased expression of the anti-apoptotic protein MCL-1. This, in turn, suppresses apoptosis and supports MDS/AML cell viability. Targeting the O-GlcNAcylated form of METTL3 using a competitive peptide significantly inhibited MDS/AML progression in preclinical models. In conclusion, our findings reveal a novel O-GlcNAcylation-dependent mechanism that regulates METTL3 stability and oncogenic activity through the m6A-SRSF1-MCL-1 axis, highlighting a potential therapeutic strategy for MDS and AML.
Insights
O-GlcNAcylation stabilizes methyltransferase-like protein 3 (METTL3), promoting myeloid leukemia survival. Targeting this modification offers a potential therapeutic strategy for myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML).
Area of Science:
- Molecular Biology
- Epigenetics
- Cancer Biology
Background:
- N6-methyladenosine (m6A) modification, regulated by methyltransferase-like protein 3 (METTL3), is crucial in RNA metabolism and leukemogenesis.
- The post-translational regulation of METTL3 stability and function, particularly in myeloid malignancies, is not fully understood.
- O-GlcNAcylation is a common post-translational modification on nuclear and cytosolic proteins.
Purpose of the Study:
- To investigate whether METTL3 undergoes O-GlcNAcylation.
- To determine the impact of METTL3 O-GlcNAcylation on its stability and oncogenic function in myeloid malignancies.
- To elucidate the molecular mechanisms by which O-GlcNAcylated METTL3 contributes to MDS and AML.
Main Methods:
- Detection of METTL3 O-GlcNAcylation in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) patient samples.
- Functional assays to assess the effect of O-GlcNAcylation on METTL3 stability and leukemic cell survival.
- Mechanistic studies involving mRNA stabilization of serine and arginine-rich splicing factor 1 (SRSF1) and expression of MCL-1.
- Preclinical models using a competitive peptide to target O-GlcNAcylated METTL3.
Main Results:
- METTL3 is O-GlcNAcylated in MDS and AML, with expression positively correlating with O-GlcNAcylation levels.
- O-GlcNAcylation enhances METTL3 protein stability and promotes leukemic cell survival.
- O-GlcNAcylated METTL3 stabilizes SRSF1 mRNA, increasing anti-apoptotic MCL-1 expression and supporting MDS/AML cell viability.
- Targeting O-GlcNAcylated METTL3 with a peptide inhibitor significantly reduced MDS/AML progression in preclinical models.
Conclusions:
- A novel O-GlcNAcylation-dependent mechanism regulates METTL3 stability and oncogenic activity in myeloid malignancies.
- The m6A-SRSF1-MCL1 axis is critical for METTL3-mediated leukemogenesis.
- Targeting O-GlcNAcylated METTL3 represents a promising therapeutic strategy for MDS and AML.
