Optimization of peptide-plasmid DNA vectors formulation for gene delivery in cancer therapy exploring design of experiments

• Published in: Colloids and surfaces, B, Biointerfaces Vol. 183; p. 110417
• Main Authors: Sousa, Ângela, Almeida, Ana M., Faria, Rúben, Konate, Karidia, Boisguerin, Prisca, Queiroz, João A., Costa, Diana
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-11-2019; Elsevier
• Subjects: Amino Acid Sequence; Cancer gene therapy; Cellular Biology; Design of experiments; Factor Analysis, Statistical; Gene delivery; Gene Transfer Techniques; Genetic Therapy - methods; Humans; Hydrogen-Ion Concentration; Life Sciences; Neoplasms - genetics; Neoplasms - therapy; Optimal peptide/pDNA formulations; pDNA condensation; peptide/pDNA vectors; Peptides - chemical synthesis; Peptides - genetics; Peptides - metabolism; Plasmids - chemistry; Plasmids - metabolism; Protein Binding; ral GTP-Binding Proteins - genetics; ral GTP-Binding Proteins - metabolism; Static Electricity; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Gene delivery; Optimal peptide/pDNA formulations; Design of experiments; pDNA condensation; peptide/pDNA vectors; Cancer gene therapy
• ISSN: 0927-7765; 1873-4367
• DOI: 10.1016/j.colsurfb.2019.110417

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•The pDNA condensation ability of RALA and WRAP5 has been evaluated.•A design of experiments tool (DoE) was used to find the optimal formulations.•The buffer pH and N/P ratio were considered the DoE inputs.•Vector size, zeta potential and pDNA complexation capacity were DoE outputs.•The ideal vectors were revealed: a great contribution for progresses in cancer therapy.
The field of gene therapy still attracts great interest due to its potential therapeutic effect towards the most deadly diseases, such as cancer. For cancer gene therapy to be feasible and viable in a clinical setting, the design and development of a suitable gene delivery system is imperative. Peptide based vectors, in particular, reveal to be promising for therapeutic gene release. Following this, two different peptides, RALA and WRAP5, have been investigated mainly regarding their ability to form complexes with a p53 encoding plasmid (pDNA) with suitable properties for gene delivery. To address this issue, and after an initial screening study focused on the dependence of pDNA complexation capacity with the nitrogen to phosphate groups (N/P) ratio, a design of experiments (DoE) tool has been employed. For each peptide/pDNA system, parameters such as, the buffer pH and the N/P ratio were considered the DoE inputs and the vector size, zeta potential and pDNA complexation capacity (CC) were monitored as DoE outputs. The main goal was to find the optimal experimental conditions to minimize particle sizes, as well as, to maximize the positive surface charges of the formulated nanosystems and maximize the pDNA CC. Through the DoE method applied, the optimal RALA/pDNA and WRAP5/pDNA formulations were revealed and show interesting features related to peptide structure and pDNA complexation ability. This work illustrates the great utility of experimental design tools in optimizing the formulation of peptide/pDNA vectors in a minimum number of experiments providing relevant knowledge for the development of more suitable and efficient gene delivery systems. The new insights achieved on these carriers clearly instigate deeper research on gene therapy.