LentiRILES, a miRNA-ON sensor system for monitoring the functionality of miRNA in cancer biology and therapy

• Published in: RNA biology Vol. 18; no. sup1; pp. 198 - 214
• Main Authors: Simion, Viorel, Loussouarn, Claire, Laurent, Yoan, Roncali, Loris, Gosset, David, Reverchon, Flora, Rousseau, Audrey, Martin, Francisco, Midoux, Patrick, Pichon, Chantal, Garcion, Emmanuel, Baril, Patrick
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 15.10.2021
• Subjects: Animals; Apoptosis; Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Biosensing Techniques - methods; Cell Cycle; Cell Movement; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Genes, Reporter; Glioblastoma - genetics; Glioblastoma - metabolism; Glioblastoma - pathology; Humans; Lentivirus - genetics; Life Sciences; Luminescent Measurements; Male; Mice; Mice, SCID; MicroRNAs - analysis; MicroRNAs - genetics; MicroRNAs - metabolism; Middle Aged; Technical Paper; Tumor Cells, Cultured; miRNA; glioblastoma; RNA therapeutics; molecular imaging; RILES
• ISSN: 1547-6286; 1555-8584
• DOI: 10.1080/15476286.2021.1978202

A major unresolved challenge in miRNA biology is the capacity to monitor the spatiotemporal activity of miRNAs expressed in animal disease models. We recently reported that the miRNA-ON monitoring system called RILES (RNAi-inducible expression Luciferase system) implanted in lentivirus expression system (LentiRILES) offers unique opportunity to decipher the kinetics of miRNA activity , in relation with their intracellular trafficking in glioblastoma cells. In this study, we describe in detail the method for the production of LentiRILES stable cell lines and employed it in several applications in the field of miRNA biology and therapy. We show that LentiRILES is a robust, highly specific and sensitive miRNA sensor system that can be used as a single-cell miRNA monitoring method, cell-based screening platform for miRNA therapeutics and as a tool to analyse the structure-function relationship of the miRNA duplex. Furthermore, we report the kinetics of miRNA activity upon the intracranial delivery of miRNA mimics in an orthotopic animal model of glioblastoma. This information is exploited to evaluate the tumour suppressive function of miRNA-200c as locoregional therapeutic modality to treat glioblastoma. Our data provide evidence that LentiRILES is a robust system, well suited to resolve the activity of endogenous and exogenously expressed miRNAs from basic research to gene and cell therapy.