Abstract
Cluster of differentiation 73 (CD73), a cell surface enzyme that catalyzes adenosine monophosphate (AMP) breakdown to adenosine, is differentially expressed in cancers and has prognostic significance. We investigated its expression profile in neuroblastoma (NB), its association with NB clinical outcomes, and its influence in the regulation of cancer stem cells’ (CSCs) stemness maintenance. RNA-Seq data mining (22 independent study cohorts, total n = 3836) indicated that high CD73 can predict good NB prognosis. CD73 expression (immunohistochemistry) gauged in an NB patient cohort (n = 87) showed a positive correlation with longer overall survival (OS, P = 0.0239) and relapse-free survival (RFS, P = 0.0242). Similarly, high CD73 correlated with longer OS and RFS in advanced disease stages, MYCN non-amplified (MYCN-na), and Stage-4-MYCN-na subsets. Despite no definite association in children < 2 years old (2Y), high CD73 correlated with longer OS (P = 0.0294) and RFS (P = 0.0315) in children > 2Y. Consistently, high CD73 was associated with better OS in MYCN-na, high-risk, and stage-4 subsets of children > 2Y. Multivariate analysis identified CD73 as an independent (P = 0.001) prognostic factor for NB. Silencing CD73 in patient-derived (stage 4, progressive disease) CHLA-171 and CHLA-172 cells revealed cell-line-independent activation of 58 CSC stemness maintenance molecules (QPCR profiling). Overexpressing CD73 in CHLA-20 and CHLA-90 cells with low CD73 and silencing in CHLA-171 and CHLA-172 cells with high CD73 showed that CD73 regulates epithelial to mesenchymal transition (E-Cadherin, N-Cadherin, Vimentin), stemness maintenance (Sox2, Nanog, Oct3/4), self-renewal capacity (Notch), and differentiation inhibition (leukemia inhibitory factor, LIF) proteins (confocal-immunofluorescence). These results demonstrate that high CD73 can predict good prognosis in NB, and further suggest that CD73 regulates stemness maintenance in cells that defy clinical therapy.
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Acknowledgements
The authors acknowledge the Stephenson Cancer Center (SCC) Cancer Tissue Pathology Core for all TMA and IHC services and the SCC Cancer Functional Genomics Core for the help with the high-content confocal immunofluorescence. The authors also acknowledge the OUHSC staff editor (Ms. Kathy Kyler) for the help in critically reviewing this manuscript.
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The National Institutes of Health, NIH-P20GM103639, and Oklahoma Center for the Advancement of Science and Technology, OCAST-HR19-045.
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NA contributed to the conception and design of the experiments. DJ, DS, SA, and NA performed the experiments and contributed to the acquisition of the data. NA, SA, and DS contributed to data analysis and interpretation of the data. NA and DJ drafted the manuscript, and SA helped in revising it critically. ZY, AE, and AB contributed to biospecimen and clinical data collection, histopathological grading, and data interpretation. All of the authors read and approved the final manuscript.
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Highlights
• CD73 abundance associated with better survival in subsets of children with high-risk (MYCN non-amplified, aged >2 years) neuroblastoma.
• Loss of CD73 in neuroblastoma contributes to the activation of EMT and stemness maintenance, and in the regulation of differentiation, the hallmarks of disease progression.
• CD73 loss independently predicts poor survival and serves as a novel prognostic tool for neuroblastoma.
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Jain, D., Somasundaram, D.B., Aravindan, S. et al. Prognostic significance of NT5E/CD73 in neuroblastoma and its function in CSC stemness maintenance. Cell Biol Toxicol 39, 967–989 (2023). https://doi.org/10.1007/s10565-021-09658-1
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DOI: https://doi.org/10.1007/s10565-021-09658-1