Nano-mediated delivery of double-stranded RNA for gene therapy of glioblastoma multiforme.

Małgorzata Grabowska,Dariusz Wawrzyniak,Katarzyna Rolle,Bartosz F Grześkowiak,Jan Barciszewski,Paweł Głodowicz,Stefan Jurga,Radosław Mrówczyński,Kosma Szutkowski,Ilya Ulasov

doi:10.1371/journal.pone.0213852

Abstract

Glioblastoma multiforme (GBM) is the most common type of malignant gliomas, characterized by genetic instability, intratumoral histopathological variability and unpredictable clinical behavior. Disappointing results in the treatment of gliomas with surgery, radiation and chemotherapy have fueled a search for new therapeutic targets and treatment modalities. Here we report new approach towards RNA interference therapy of glioblastoma multiforme based on the magnetic nanoparticles delivery of the double-stranded RNA (dsRNA) with homological sequences to mRNA of tenascin-C (TN-C), named ATN-RNA. The obtained nanocomposite consisted of polyethyleneimine (PEI) coated magnetic nanoparticles conjugated to the dsRNA show high efficiency in ATN-RNA delivery, resulting not only in significant TN-C expression level suppressesion, but also impairing the tumor cells migration. Moreover, synthesized nanomaterials show high contrast properties in magnetic resonance imaging (MRI) and low cytotoxicity combining with lack of induction of interferon response. We believe that the present work is a successful combination of effective, functional, non-immunostimulatory dsRNA delivery system based on magnetic nanoparticles with high potential for further application in GBM therapy.

Highlights

Encouraged by the results obtained from zeta potential measurement we investigated the colloidal stability of Magnetic nanoparticles coated with PEI (Mag@PEI) NPs in real-time by Multiple Light Scattering (MLS) using Turbiscan apparatus
We have successfully demonstrated a new double-stranded RNA (dsRNA) delivery system that harnesses well- characterized magnetic nanoparticles coated with PEI to effectively silence expression of tenascin-C in glioblastoma multiforme cell line
The most important thing is that synthesized nanocarrier was a more efficient tool in delivery ATN-RNA than routinely used Lipofectamine In consequence, the gene therapy employing ATN was improved resulting in stronger silencing of TN-C, followed by the further diminishing of migration of glioblastoma cells

Summary

Introduction

The part of MRI studies was performed in the frame of the project number 2014/13/D/ST5/02793 supported by the National Science Centre, Poland to RM. Accounting for less than 2% of adult cancers, gliomas are the most common form of malignant primary brain tumor in adults.[1] Glioblastoma multiforme constitutes 25% of all malignant nervous system tumors and the median overall survival remains around 12–15 months, even after combination treatments of cytoreductive surgical resection, radiotherapy, and adjuvant oral chemotherapy with temozolomide[2,3,4,5] The recent medical treatment strategies have been progressing toward individualized therapy and many targeted drugs have. Nano-mediated delivery of double-stranded RNA to glioblastoma cells to the Foundation for Polish Science (FNP) for its support by Stypendium START scholarship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Methods

Results

Conclusion