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

被引：5

作者：

Boissinot, M. ^{[1
]}

King, H. ^{[2
]}

Adams, M. ^{[2
]}

Higgins, J. ^{[1
]}

Shaw, G. ^{[1
]}

Ward, T. A. ^{[1
]}

Steele, L. P. ^{[1
]}

Tams, D. ^{[1
,3
]}

Morton, R. ^{[4
]}

Polson, E. ^{[4
]}

da Silva, B. ^{[4
]}

Droop, A. ^{[5
]}

Hayes, J. L. ^{[1
,6
]}

Martin, H. ^{[7
]}

Laslo, P. ^{[8
]}

Morrison, E. ^{[9
]}

Tomlinson, D. C. ^{[2
,7
]}

Wurdak, H. ^{[4
]}

Bond, J. ^{[2
,10
]}

Lawler, S. E. ^{[1
,11
]}

Short, S. C. ^{[1
,12
,13
]}

机构：

[1] Univ Leeds, St Jamess Hosp, Leeds Inst Med Res, Radiat Biol & Therapy Grp, Leeds LS9 7TF, W Yorkshire, England

[2] Univ Leeds, St Jamess Hosp, BioScreening Technol Grp, Leeds LS9 7TF, W Yorkshire, England

[3] Babraham, Roslin Cell Sci, Cambridge CB22 3AT, England

[4] Univ Leeds, St Jamess Hosp, Leeds Inst Med Res, Stem Cells & Brain Tumour Res Grp, Leeds LS9 7TF, W Yorkshire, England

[5] Univ Leeds, MRC Med Bioinformat Ctr, Clarendon Way, Leeds LS2 9NL, W Yorkshire, England

[6] Univ Calif Berkeley, Sch Publ Hlth, Berkeley, CA 94720 USA

[7] Univ Leeds, Fac Biol Sci, Sch Mol & Cellular Biol, Leeds LS2 9JT, W Yorkshire, England

[8] Univ Leeds, St Jamess Hosp, Leeds Inst Med Res, Myeloid Differentiat Grp, Leeds LS9 7TF, W Yorkshire, England

[9] Univ Leeds, St Jamess Hosp, Leeds Inst Med Res, Cell Biol Res Grp, Leeds LS9 7TF, W Yorkshire, England

[10] Univ Leeds, St Jamess Hosp, Leeds Inst Med Res, Microcephaly & Neurogenesis Res Grp, Leeds LS9 7TF, W Yorkshire, England

[11] Harvard Med Sch, Brigham & Womens Hosp, Dept Neurosurg, Harvey Cushing Neurooncol Labs, Boston, MA 02115 USA

[12] Univ Leeds, St Jamess Hosp, St Jamess Inst Oncol, Leeds LS9 7TF, W Yorkshire, England

[13] Univ Leeds, St Jamess Hosp, Leeds Inst Med Res, Leeds LS9 7TF, W Yorkshire, England

来源：

ONCOGENE | 2020年 / 39卷 / 30期

基金：

英国医学研究理事会;

关键词：

CLEAVAGE FURROW FORMATION; EXCHANGE FACTORS; ECT2; TARGETS; POLYPLOIDIZATION; PATHWAYS; MIRBASE; MEDIATE; TRIO;

D O I：

10.1038/s41388-020-1360-y

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 similar to 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.

引用

页码：5292 / 5306

页数：15

共 48 条

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