Abstract
Delivery of pharmaceutical agents across a blood–brain barrier (BBB) is a challenge for brain cancer therapy. In this study, an in vitro BBB model was utilized to study the delivery of oligonucleotides across brain endothelial cells targeting to glioma cells in a Transwell™ setup. A series of novel peptides were synthesized by covalent conjugation of cell-penetrating peptides with targeting peptides for delivery of gene-based therapeutics. These peptides were screened for passage across the Transwell™ and we found the most efficient peptide PepFect32 from originating PepFect 14 coupled with the targeting peptide angiopep-2. PepFect32/pDNA nanocomplexes exhibited high transcytosis across the BBB in vitro model and the highest transfection efficiency to glioma cells. In conclusion, PepFect32 revealed the most efficient peptide-based vector for pDNA delivery across in vitro BBB model.
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Abbreviations
- CPP:
-
Cell-penetrating peptide
- BBB:
-
Blood–brain barrier
- ANG:
-
Angiopep-2
- gHo:
-
Glioma-homing peptide
- CR:
-
Charge ratio
- TEER:
-
Transepithelial electrical resistance
- DIPEA:
-
Diisopropylethylamine
- TFA:
-
Trifluoroacetic acid
- TIS:
-
Triisopropylsilane
- pGL3:
-
Luciferase-encoding plasmid
- DLS:
-
Dynamic light scattering
- TAE:
-
Tris–acetate–EDTA
- EtBr:
-
Ethidium bromide
- RLU:
-
Relative light unit
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Acknowledgments
This work was supported by the Office of the High Education Commission and Mahidol University under the National Research Universities Initiative, Thailand; by the EU through the European Development Fund through the Center of Excellence in Chemical Biology, Estonia; by the targeted financing SF0180027s08 from the Estonian Government; by the Innovative Medicines Initiative Joint Undertaking under grant agreement n° 115363 resources of which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/2007-2013) (IMI), Swedish Cancer Foundation, Sweden. We kindly thank Carmine Pasquale Cerrato for figure modification of the in vitro BBB model.
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Srimanee, A., Regberg, J., Hallbrink, M. et al. Peptide-Based Delivery of Oligonucleotides Across Blood–Brain Barrier Model. Int J Pept Res Ther 20, 169–178 (2014). https://doi.org/10.1007/s10989-013-9378-4
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DOI: https://doi.org/10.1007/s10989-013-9378-4