原创力文档reduced expression of brain-enriched micrornas in glioblastomas permits targeted regulation of a cell death gene减少表达brain-enriched microrna在恶性胶质瘤允许有针对性的调控的细胞死亡的基因.pdfVIP
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reduced expression of brain-enriched micrornas in glioblastomas permits targeted regulation of a cell death gene减少表达brain-enriched microrna在恶性胶质瘤允许有针对性的调控的细胞死亡的基因
Reduced Expression of Brain-Enriched microRNAs in
Glioblastomas Permits Targeted Regulation of a Cell
Death Gene
Rebecca L. Skalsky, Bryan R. Cullen*
Department of Molecular Genetics and Microbiology and Center for Virology, Duke University Medical Center, Durham, North Carolina, United States of America
Abstract
Glioblastoma is a highly aggressive malignant tumor involving glial cells in the human brain. We used high-throughput
sequencing to comprehensively profile the small RNAs expressed in glioblastoma and non-tumor brain tissues. MicroRNAs
(miRNAs) made up the large majority of small RNAs, and we identified over 400 different cellular pre-miRNAs. No known
viral miRNAs were detected in any of the samples analyzed. Cluster analysis revealed several miRNAs that were significantly
down-regulated in glioblastomas, including miR-128, miR-124, miR-7, miR-139, miR-95, and miR-873 . Post-transcriptional
editing was observed for several miRNAs, including the miR-376 family, miR-411, miR-381, and miR-379. Using the deep
sequencing information, we designed a lentiviral vector expressing a cell suicide gene, the herpes simplex virus thymidine
kinase (HSV-TK) gene, under the regulation of a miRNA, miR-128, that was found to be enriched in non-tumor brain tissue
yet down-regulated in glioblastomas, Glioblastoma cells transduced with this vector were selectively killed when cultured in
the presence of ganciclovir. Using an in vitro model to recapitulate expression of brain-enriched miRNAs, we demonstrated
that neuronally differentiated SH-SY5Y cells transduced with the miRNA-regulated HSV-TK vector are protected from killing
by expression of endogenous miR-128. Together, these results provide an in-depth analysis of miRNA dysregulation in
glioblastoma and demonstrate the potential utility of these data in the design of miRNA-regulated therapies for
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