Development and characterization of CD73-siRNA-loaded nanoemulsion: effect on C6 glioma cells and primary astrocytes

Abstract

Introduction: Glioblastoma (GB) is the most common malignant brain tumor and is characterized by high invasiveness, poor prognosis, and limited therapeutic options. Silencing gene expression, through the use of small interfering RNA (siRNA), has been proposed as an alternative to conventional cancer therapy. Here, we evaluated the potential of CD73 as a new therapeutic target, since it is overexpressed in solid tumors and has emerged as a promising target to control GB progression.

Methods: A cationic nanoemulsion (NE) as an intravenous siRNA-CD73 delivery system was developed and its effect on C6 glioma cell viability was determined.

Results: The nanostructured system was effective in complexing oligonucleotides for delivery to target cells. In addition, we observed that the NE-siRNA-CD73 complex was effective in reducing CD73 protein levels and AMPase activity, which were related to decreased C6 glioma cell viability.

Conclusions: These findings indicate the potential of siRNA-CD73-loaded cationic NE as a therapeutic alternative for glioma treatment.

Keywords:

• Glioblastoma
• ecto-5′-nucleotidase
• CD73
• siRNA
• cationic nanoemulsion
• drug delivery

Disclosure statement

No potential conflict of interest was reported by the authors.

Authors’ contribution

Fernanda C. Teixeira is responsible for experimental design, culture preparation and treatment, cell viability, discussion of results, and manuscript preparation. Fernanda Bruxel carried out nanoemulsion preparation and characterization. Juliana H. Azambuja is responsible for experimental design, culture preparation and treatment, astrocytes culture, discussion of results, and manuscript preparation. Alexandre M. Berenguer is responsible for siRNA design and discussion of the results. Marco A. Stefani is responsible for experimental design and discussion of the results. Jean Sévigny is responsible for experimental design and antibody preparation. Roselia M. Spanevello and Ana M. O. Battastini are responsible for experimental design and interpretation and discussion of the results. Helder F. Teixeira is responsible for experimental design, obtention of nanoemulsion samples, discussion of results, and manuscript preparation. Elizandra Braganhol supervised and concept the study; she is responsible for experimental design, statistical analysis, interpretation and discussion of results, and manuscript preparation.

Additional information

Funding

This work was supported by the Brazilian funding agencies: Fundo de Incentivo a Pesquisa e Eventos (FIPE-HCPA – Proc. 11-0167); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq: Processo 422298/2016-6; 312187/2018-1), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES: Processo 01), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS – Proc. 12/2011-7; 16/2551-0000265-7; PRONEX – 16/2551-0000473-0). F.C.T. was a recipient of a Master Scholarship from Capes; J.H.A. was recipient of PhD Scholarship from UFCSPA. J.S. received support from the Canadian Institutes of Health Research (CIHR) and was the recipient of a ‘Chercheur National’ Scholarship from the Fonds de Recherche du Québec – Santé (FRQS).