The splicing modulator CLK1 is a candidate oncogenic dependency in pediatric high-grade gliomas

Naqvi, A. S.; Corbett, R. J.; Seghal, P.; Conkrite, K. L.; Rathi, K. S.; Ennis, B. M.; Hayer, K. E.; Zhang, B.; Brown, M. A.; Miller, D. P.; Kraya, A.; Dybas, J. M.; Geng, Z.; Blackden, C.; Arif, S.; Chroni, A.; Lahiri, A.; Hollawell, M. L.; Storm, P. B.; Foster, J. B.; Koptyra, M.; Madsen, P. J.; Diskin, S. J.; Thomas-Tikhonenko, A.; Resnick, A. C.; Rokita, J. L.

doi:10.1101/2024.08.03.606419

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bioRxiv · 10.1101/2024.08.03.606419

The splicing modulator CLK1 is a candidate oncogenic dependency in pediatric high-grade gliomas

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Abstract

BackgroundPediatric brain tumors are the leading cause of disease-related mortality in children, yet many aggressive tumors lack effective therapies. RNA splicing is a hallmark of cancer, but it has not yet been systematically studied in pediatric brain tumors. MethodsWe analyzed 729 pediatric brain tumors spanning histologies and molecular subtypes to quantify differential tumor splicing. We developed the Splicing Burden Index (SBI) to enable cross-sample comparisons and performed hierarchical clustering of highly variable splice events to define splicing-informed tumor groups. These were integrated with clinical outcomes, pathway activity, and proteogenomic data. Recurrent splice events were prioritized for predicted functional impact, and in vitro perturbation studies were performed targeting the splicing kinase CDC-like kinase 1 (CLK1). ResultsSBI revealed substantial inter- and intra-histology heterogeneity. Clusters were enriched for histologies and molecular subtypes, several of which were independently associated with survival beyond histology and clinical covariates. Spliceosome pathway activity varied across clusters and was associated with worse survival, yet was not correlated with SBI, indicating distinct dimensions of splicing dysregulation. Functional prioritization identified a recurrent in CLK1 exon 4, required for canonical kinase activity. CLK1 exon 4 inclusion followed an oncofetal pattern and showed context-dependent associations with outcome distinct from total CLK1 expression. Pharmacologic inhibition and exon 4-specific perturbation of CLK1 reduced tumor cell viability and disrupted cancer-relevant splicing and transcriptional programs. ConclusionsThis study systematically characterizes splicing in pediatric brain tumors, identifies splicing-informed subgroups, and prioritizes CLK1 exon 4 as an oncofetal tumor-specific event, motivating further preclinical exploration. Key Points[bullet] Splicing analysis of 729 pediatric CNS tumors identifies splicing-defined clusters. [bullet]CLK1 exon 4 inclusion is widespread and developmentally regulated. [bullet]Exon-level CLK1 regulation shows context-dependent links to prognosis in aggressive CNS tumors.

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Naqvi, A. S., Corbett, R. J., Seghal, P., Conkrite, K. L., Rathi, K. S., Ennis, B. M., Hayer, K. E., Zhang, B., Brown, M. A., Miller, D. P., Kraya, A., Dybas, J. M., Geng, Z., Blackden, C., Arif, S., Chroni, A., Lahiri, A., Hollawell, M. L., Storm, P. B., Foster, J. B., Koptyra, M., Madsen, P. J., Diskin, S. J., Thomas-Tikhonenko, A., Resnick, A. C., Rokita, J. L.. 2024-08-06. The splicing modulator CLK1 is a candidate oncogenic dependency in pediatric high-grade gliomas. https://doi.org/10.1101/2024.08.03.606419

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