Profiling cytotoxic microRNAs in pediatric and adult glioblastoma cells by high-content screening, identification, and validation of miR-1300
| Autor: | Henry King, Alastair Droop, Ewan E. Morrison, Ruth Morton, Susan C Short, Julie Higgins, Gary Shaw, Peter Laslo, B. da Silva, Euan S. Polson, Lynette Steele, Daniel Tams, Heather L. Martin, Matthew Adams, Heiko Wurdak, Darren C. Tomlinson, Sean E. Lawler, Marjorie Boissinot, Josie Hayes, Jacquelyn Bond, Thomas Ward |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Adult Cancer Research Cell Survival Megakaryocyte differentiation Mitosis Biology Article Non-coding RNAs 03 medical and health sciences 0302 clinical medicine Cell Line Tumor Proto-Oncogene Proteins microRNA Genetics Cytotoxic T cell Humans Child Molecular Biology 3' Untranslated Regions Oncogene Brain Neoplasms Gene Expression Profiling Cell Differentiation Transfection Oncogenes High-Throughput Screening Assays Gene Expression Regulation Neoplastic MicroRNAs 030104 developmental biology 030220 oncology & carcinogenesis High-content screening Cancer research Ectopic expression Glioblastoma Megakaryocytes Cytokinesis |
| Zdroj: | Oncogene |
| ISSN: | 1476-5594 0950-9232 |
| Popis: | MicroRNAs play an important role in the regulation of mRNA translation and have therapeutic potential in cancer and other diseases. To profile the landscape of microRNAs with significant cytotoxicity in the context of glioblastoma (GBM), we performed a high-throughput screen in adult and pediatric GBM cells using a synthetic oligonucleotide library representing all known human microRNAs. Bioinformatics analysis was used to refine this list and the top seven microRNAs were validated in a larger panel of GBM cells using state-of-the-art in vitro assays. The cytotoxic effect of our most relevant candidate was assessed in a preclinical model. Our screen identified ~100 significantly cytotoxic microRNAs with 70% concordance between cell lines. MicroRNA-1300 (miR-1300) was the most potent and robust candidate. We observed a striking binucleated phenotype in miR-1300 transfected cells due to cytokinesis failure followed by apoptosis. This was also observed in two stem-like patient-derived cultures. We identified the physiological role of miR-1300 as a regulator of endomitosis in megakaryocyte differentiation where blockade of cytokinesis is an essential step. In GBM cells, where miR-1300 is normally not expressed, the oncogene Epithelial Cell Transforming 2 (ECT2) was validated as a direct key target. ECT2 siRNA phenocopied the effects of miR-1300, and ECT2 overexpression led to rescue of miR-1300 induced binucleation. We showed that ectopic expression of miR-1300 led to decreased tumor growth in an orthotopic GBM model. Our screen provides a resource for the neuro-oncology community and identified miR-1300 as a novel regulator of endomitosis with translatable potential for therapeutic application. |
| Databáze: | OpenAIRE |
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