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

RNA Biol. 2021 Oct 15;18(sup1):198-214. doi: 10.1080/15476286.2021.1978202. Epub 2021 Sep 27.

Abstract

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 vitro, 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 in vitro 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.

Keywords: RILES; RNA therapeutics; glioblastoma; miRNA; molecular imaging.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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*
  • Luminescent Measurements
  • Male
  • Mice
  • Mice, SCID
  • MicroRNAs / analysis*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism
  • Middle Aged
  • Tumor Cells, Cultured

Substances

  • Biomarkers, Tumor
  • MIRN200 microRNA, human
  • MicroRNAs

Grants and funding

This work was funded by the Institut National du Cancer (INCa) [PL-BIO 2014–2020, consortium MARENGO: MicroRNA agonist and antagonist Nanomedicines for GliOblastoma treatment: from molecular programming to preclinical validation to P.B and E.G]; La Ligue Contre le Cancer [Région du Loiret 2017-2019 to P.B]; ARD 2020 Biopharmaceuticals [RNA cell factory 2017-2020 to C.P]. The work was also related to the French National Research Agency (ANR) through the LabEx IRON (Innovative Radiopharmaceuticals in Oncology and Neurology to E.G) as part of the French government Investissements d’Avenir program (ANR-11-LABX-0018 to E.G) and to the ANR under the frame of EuroNanoMed III (project GLIOSILK to E.G). LR was a PhD fellow funded by the LabEx IRON-2 and the University of Angers. h[;