Affordable Access

deepdyve-link
Publisher Website

Profiling cytotoxic microRNAs in pediatric and adult glioblastoma cells by high-content screening, identification, and validation of miR-1300

Authors
  • Boissinot, M.1
  • King, H.1
  • Adams, M.2
  • Higgins, J.2
  • Shaw, G.1
  • Ward, T. A.1
  • Steele, L. P.1
  • Tams, D.1, 3
  • Morton, R.1
  • Polson, E.4
  • da Silva, B.4
  • Droop, A.5
  • Hayes, J. L.1, 6
  • Martin, H.5
  • Laslo, P.7
  • Morrison, E.8
  • Tomlinson, D. C.2, 5
  • Wurdak, H.4
  • Bond, J.2, 9
  • Lawler, S. E.1, 10
  • And 1 more
  • 1 Radiation Biology and Therapy Group, Leeds Institute of Medical Research, University of Leeds, St James’s Hospital, Leeds, LS9 7TF UK
  • 2 BioScreening Technology Group, University of Leeds, St James’s Hospital, Leeds, LS9 7TF UK
  • 3 Roslin Cell Sciences, Babraham, Cambridge, CB22 3AT UK
  • 4 Stem Cells and Brain Tumour Research Group, Leeds Institute of Medical Research, University of Leeds, St James’s Hospital, Leeds, LS9 7TF UK
  • 5 University of Leeds,
  • 6 University of California Berkeley,
  • 7 Myeloid Differentiation Group, Leeds Institute of Medical Research, University of Leeds, St James’s Hospital, Leeds, LS9 7TF UK
  • 8 Cell Biology Research Group, Leeds Institute of Medical Research, University of Leeds, St James’s Hospital, Leeds, LS9 7TF UK
  • 9 Microcephaly and Neurogenesis Research Group, Leeds Institute of Medical Research, University of Leeds, St James’s Hospital, Leeds, LS9 7TF UK
  • 10 Harvey Cushing Neurooncology Laboratories, Department of Neurosurgery, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115 USA
  • 11 St James’s Institute of Oncology and Leeds Institute of Medical Research, University of Leeds, St James’s Hospital, Leeds, LS9 7TF UK
Type
Published Article
Journal
Oncogene
Publisher
Nature Publishing Group UK
Publication Date
Jun 17, 2020
Volume
39
Issue
30
Pages
5292–5306
Identifiers
DOI: 10.1038/s41388-020-1360-y
PMID: 32555332
PMCID: PMC7378045
Source
PubMed Central
Keywords
License
Unknown

Abstract

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.

Report this publication

Statistics

Seen <100 times