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Spliceosomal vulnerability of MYCN-amplified neuroblastoma is contingent on PRMT5-mediated regulation of epitranscriptomic and metabolomic pathways
Jodie Bojko 
,
Madhu Kollareddy ,
Marianna Szemes,
Jacob Bellamy ,
Evon Poon ,
Ahmad Moukachar,
Danny Legge,
Emma E. Vincent ,
Nick Jones ,
Sally Malik ,
Alexander Greenhough ,
Alex Paterson ,
Ji Hyun Park,
Kelli Gallacher ,
Louis Chesler,
Karim Malik
,
Madhu Kollareddy ,
Marianna Szemes,
Jacob Bellamy ,
Evon Poon ,
Ahmad Moukachar,
Danny Legge,
Emma E. Vincent ,
Nick Jones ,
Sally Malik ,
Alexander Greenhough ,
Alex Paterson ,
Ji Hyun Park,
Kelli Gallacher ,
Louis Chesler,
Karim Malik
Cancer Letters, Volume: 604, Start page: 217263
Swansea University Author:
Nick Jones
Full text not available from this repository: check for access using links below.
DOI (Published version): 10.1016/j.canlet.2024.217263
Abstract
Approximately 50 % of poor prognosis neuroblastomas arise due to MYCN over-expression. We previously demonstrated that MYCN and PRMT5 proteins interact and PRMT5 knockdown led to apoptosis of MYCN-amplified (MNA) neuroblastoma. Here we evaluate the highly selective first-in-class PRMT5 inhibitor GSK...
| Published in: | Cancer Letters |
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| ISSN: | 0304-3835 |
| Published: |
Elsevier BV
2024
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| Online Access: |
Check full text
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| URI: | https://cronfa.swan.ac.uk/Record/cronfa67770 |
| Abstract: |
Approximately 50 % of poor prognosis neuroblastomas arise due to MYCN over-expression. We previously demonstrated that MYCN and PRMT5 proteins interact and PRMT5 knockdown led to apoptosis of MYCN-amplified (MNA) neuroblastoma. Here we evaluate the highly selective first-in-class PRMT5 inhibitor GSK3203591 and its in vivo analogue GSK3326593 as targeted therapeutics for MNA neuroblastoma. Cell-line analyses show MYCN-dependent growth inhibition and apoptosis, with approximately 200-fold greater sensitivity of MNA neuroblastoma lines. RNA sequencing of three MNA neuroblastoma lines treated with GSK3203591 reveal deregulated MYCN transcriptional programmes and altered mRNA splicing, converging on key regulatory pathways such as DNA damage response, epitranscriptomics and cellular metabolism. Stable isotope labelling experiments in the same cell lines demonstrate that glutamine metabolism is impeded following GSK3203591 treatment, linking with disruption of the MLX/Mondo nutrient sensors via intron retention of MLX mRNA. Interestingly, glutaminase (GLS) protein decreases after GSK3203591 treatment despite unchanged transcript levels. We demonstrate that the RNA methyltransferase METTL3 and cognate reader YTHDF3 proteins are lowered following their mRNAs undergoing GSK3203591-induced splicing alterations, indicating epitranscriptomic regulation of GLS; accordingly, we observe decreases of GLS mRNA m6A methylation following GSK3203591 treatment, and decreased GLS protein following YTHDF3 knockdown. In vivo efficacy of GSK3326593 is confirmed by increased survival of Th-MYCN mice, with drug treatment triggering splicing events and protein decreases consistent with in vitro data. Together our study demonstrates the PRMT5-dependent spliceosomal vulnerability of MNA neuroblastoma and identifies the epitranscriptome and glutamine metabolism as critical determinants of this sensitivity. |
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| College: |
Faculty of Medicine, Health and Life Sciences |
| Funders: |
he authors would like to thank Cancer Research UK (A12743/A21046), Neuroblastoma UK, the Biotechnology and Biological Sciences Research Council (BB/P008232/1), and Children's Cancer and Leukaemia Group (CCLG) and The Little Princess Trust (LPT) for funding this study. E.P. and L.C. were supported by Cancer Research UK Programme Award A28278 and ICR institutional funding. A.G. is funded by the Wallscourt Foundation. |
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