Abstract
Non-viral transfection vectors are commonly used for oligonucleotide (ON) delivery but face many challenges before reaching the desired compartments inside cells. With the support of additional compounds, it might be more feasible for a vector to endure the barriers and achieve efficient delivery. In this report, we screened 18 different excipients and evaluated their effect on the performance of peptide dendrimer/lipid vector to deliver single-stranded, splice-switching ONs under serum conditions. Transfection efficiency was monitored in four different reporter cell lines by measuring splice-switching activity on RNA and protein levels. All reporter cell lines used had a mutated human β-globin intron 2 sequence interrupting the luciferase gene, which led to an aberrant splicing of luciferase pre-mRNA and subsidence of luciferase protein translation. In the HeLa Luc/705 reporter cell line (a cervical cancer cell line), the lead excipients (Polyvinyl derivatives) potentiated the splice-switching activity up to 95-fold, compared to untreated cells with no detected cytotoxicity. Physical characterization revealed that lead excipients decreased the particle size and the zeta potential of the formulations. In vivo biodistribution studies emphasized the influence of formulations as well as the type of excipients on biodistribution profiles of the ON. Subsequently, we suggest that the highlighted impact of tested excipients would potentially assist in formulation development to deliver ON therapeutics in pre-clinical and clinical settings.
Highlights
Peptide dendrimers were first reported in early 1980s by Denkewalter et al [1] and Aharoni et al [2] and while they are nowadays categorized into different classes, full peptide dendrimers are the most interesting, from the delivery point of view
We previously reported the use of sucrose excipient that enhanced serum transfection efficacy of Peptide dendrimer/lipid/ON (PDLO)-complexes to a level comparable to Lipofectamine 2000 (L2000) [17]
Splice-correction levels were comparable to L2000 upon transfection with formulations supplemented with lactose, galactosamine, or N-acetyl galactosamine with 27, 29, 26-fold higher luciferase activity than untreated cells, respectively
Summary
Peptide dendrimers were first reported in early 1980s by Denkewalter et al [1] and Aharoni et al [2] and while they are nowadays categorized into different classes, full peptide dendrimers are the most interesting, from the delivery point of view. This class is composed of only amino acids, from the core to the outer shell [3]. The replacement of polymeric dendrimers with peptide dendrimers result in reduced cytotoxicity, they still face many biological barriers for effective delivery. The vector combination strategy has been successfully applied in a number of studies for the delivery of different cargos [8,9,10,11,12,13]
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call