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Essential role of METTL3-mediated m6A modification in glioma stem-like cells maintenance and radioresistance

Oncogene volume 37, pages 522–533 (2018)Cite this article

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Abstract

Despite advances in biology and therapeutic modalities, existence of highly tumorigenic glioma stem-like cells (GSCs) makes glioblastomas (GBMs) invincible. N6-methyl adenosine (m6A), one of the abundant mRNA modifications catalyzed by methyltransferase-like 3 and 14 (METTL3/14), influences various events in RNA metabolism. Here, we report the crucial role of METTL3-mediated m6A modification in GSC (neurosphere) maintenance and dedifferentiation of glioma cells. METTL3 expression is elevated in GSC and attenuated during differentiation. RNA immunoprecipitation studies identified SOX2 as a bonafide m6A target of METTL3 and the m6A modification of SOX2 mRNA by METTL3 enhanced its stability. The exogenous overexpression of 3′UTR-less SOX2 significantly alleviated the inhibition of neurosphere formation observed in METTL3 silenced GSCs. METTL3 binding and m6A modification in vivo required intact three METTL3/m6A sites present in the SOX2-3′UTR. Further, we found that the recruitment of Human antigen R (HuR) to m6A-modified RNA is essential for SOX2 mRNA stabilization by METTL3. In addition, we found a preferential binding by HuR to the m6A-modified transcripts globally. METTL3 silenced GSCs showed enhanced sensitivity to γ-irradiation and reduced DNA repair as evidenced from the accumulation of γ-H2AX. Exogenous overexpression of 3′UTR-less SOX2 in METTL3 silenced GSCs showed efficient DNA repair and also resulted in the significant rescue of neurosphere formation from METTL3 silencing induced radiosensitivity. Silencing METTL3 inhibited RasV12 mediated transformation of mouse immortalized astrocytes. GBM tumors have elevated levels of METTL3 transcripts and silencing METTL3 in U87/TIC inhibited tumor growth in an intracranial orthotopic mouse model with prolonged mice survival. METTL3 transcript levels predicted poor survival in GBMs which are enriched for GSC-specific signature. Thus our study reports the importance of m6A modification in GSCs and uncovers METTL3 as a potential molecular target in GBM therapy.

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Acknowledgements

The results published here are in part based upon data generated by The Cancer Genome Atlas (TCGA) pilot project established by the NCI and NHGRI. Information about TCGA and the investigators and institutions, which constitute the TCGA research network, can be found at http://cancergenome.nih.gov/. The use of data sets from Institute NC, GSE76976 and GSE22866 is acknowledged. We thank Dr Hiroaki Wakimoto and Dr Samuel Rabkin for providing GSCs and Dr Kenneth Kousik for 3′UTR-SOX2-Luc construct. KS thanks DST and DBT, Government of India for financial support. KS is a JC Bose Fellow of the Department of Science and Technology. AV acknowledges IISc for the fellowship.

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Authors and Affiliations

  1. Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, India

    A Visvanathan, V Patil, A Arora & K Somasundaram

  2. Sri Satya Sai Institute of Higher Medical Sciences, Bangalore, India

    A S Hegde

  3. Department of Neurosurgery, National Institute of Mental Health and Neurosciences, Bangalore, India

    A Arivazhagan

  4. Department of Neuropathology, National Institute of Mental Health and Neurosciences, Bangalore, India

    V Santosh

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Correspondence to K Somasundaram.

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Visvanathan, A., Patil, V., Arora, A. et al. Essential role of METTL3-mediated m6A modification in glioma stem-like cells maintenance and radioresistance. Oncogene 37, 522–533 (2018). https://doi.org/10.1038/onc.2017.351

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