Development of surface modified biodegradable polymeric nanoparticles to deliver GSE24.2 peptide to cells: A promising approach for the treatment of defective telomerase disorders

• Published in: European journal of pharmaceutics and biopharmaceutics Vol. 91; pp. 91 - 102
• Main Authors: Egusquiaguirre, Susana P., Manguán-García, Cristina, Pintado-Berninches, Laura, Iarriccio, Laura, Carbajo, Daniel, Albericio, Fernando, Royo, Miriam, Pedraz, José Luís, Hernández, Rosa M., Perona, Rosario, Igartua, Manuela
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2015
• Subjects: Animals; Biocompatible Materials - adverse effects; Biocompatible Materials - chemistry; Biodegradable nanoparticles; Biological Transport; Cell Cycle Proteins - administration & dosage; Cell Cycle Proteins - adverse effects; Cell Cycle Proteins - chemistry; Cell Cycle Proteins - genetics; Cell Line; Cell Survival - drug effects; Cell-penetrating peptides; Cell-Penetrating Peptides - adverse effects; Cell-Penetrating Peptides - chemistry; Cells, Cultured; Chemical Phenomena; Delayed-Action Preparations - administration & dosage; Delayed-Action Preparations - adverse effects; Delayed-Action Preparations - chemistry; Delayed-Action Preparations - therapeutic use; Drug Compounding; Drug Delivery Systems - adverse effects; Drug Liberation; Drug Stability; Dyskeratosis congenita; Dyskeratosis Congenita - drug therapy; Enzyme Reactivators - administration & dosage; Enzyme Reactivators - adverse effects; Enzyme Reactivators - chemistry; Enzyme Reactivators - therapeutic use; GSE24.2 peptide; Humans; Lactic Acid - adverse effects; Lactic Acid - chemistry; Mice; Nanoparticles - adverse effects; Nanoparticles - chemistry; Nuclear Proteins - administration & dosage; Nuclear Proteins - adverse effects; Nuclear Proteins - chemistry; Nuclear Proteins - genetics; Peptide Fragments - administration & dosage; Peptide Fragments - adverse effects; Peptide Fragments - chemistry; Peptide Fragments - genetics; Polyamines - adverse effects; Polyamines - chemistry; Polycations; Polyethylene Glycols - adverse effects; Polyethylene Glycols - chemistry; Polyglactin 910 - adverse effects; Polyglactin 910 - chemistry; Polyglycolic Acid - adverse effects; Polyglycolic Acid - chemistry; Protein Stability; Recombinant Proteins - administration & dosage; Recombinant Proteins - adverse effects; Recombinant Proteins - chemistry; Recombinant Proteins - therapeutic use; Telomerase - chemistry; Telomerase - deficiency; Telomerase - metabolism; Telomerase reactivation; Dyskeratosis congenita; GSE24.2 peptide; Cell-penetrating peptides; Biodegradable nanoparticles; Polycations; Telomerase reactivation
• ISSN: 0939-6411; 1873-3441
• DOI: 10.1016/j.ejpb.2015.01.028

[Display omitted] •Cationic and CPP-conjugated PLGA NPs encapsulating GSE24.2 peptide were developed.•PEI-PLGA, CPP5-PLGA and CPP6-PLGA-PEG NPs rescued the DNA damage in F9A353V cells.•These three formulations were able to recover the telomerase activity in VA13 cells.•These formulations are a promising approach to treat defective telomerase disorders. The aim of the present study was to develop a novel strategy to deliver intracellularly the peptide GSE24.2 for the treatment of Dyskeratosis congenita (DC) and other defective telomerase disorders. For this purpose, biodegradable polymeric nanoparticles using poly(lactic-co-glycolic acid) (PLGA NPs) or poly(lactic-co-glycolic acid)-poly ethylene glycol (PLGA-PEG NPs) attached to either polycations or cell-penetrating peptides (CPPs) were prepared in order to increase their cellular uptake. The particles exhibited an adequate size and zeta potential, with good peptide loading and a biphasic pattern obtained in the in vitro release assay, showing an initial burst release and a later sustained release. GSE24.2 structural integrity after encapsulation was assessed using SDS–PAGE, revealing an unaltered peptide after the NPs elaboration. According to the cytotoxicity results, cell viability was not affected by uncoated polymeric NPs, but the incorporation of surface modifiers slightly decreased the viability of cells. The intracellular uptake exhibited a remarkable improvement of the internalization, when the NPs were conjugated to the CPPs. Finally, the bioactivity, addressed by measuring DNA damage rescue and telomerase reactivation, showed that some formulations had the lowest cytotoxicity and highest biological activity. These results proved that GSE24.2-loaded NPs could be delivered to cells, and therefore, become an effective approach for the treatment of DC and other defective telomerase syndromes.