Two peptides targeting endothelial receptors are internalized into murine brain endothelial cells
| Autor: | Andrew J. Urquhart, Sara Björk Sigurdardóttir, Sarah Christine Christensen, Thomas Lars Andresen, Morten Nielsen, Diána Hudecz, Casper Hempel |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Central Nervous System
Endocytic cycle Peptide Nervous System Epithelium Mass Spectrometry Analytical Chemistry Mice Spectrum Analysis Techniques Drug Delivery Systems Animal Cells Medicine and Health Sciences Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry Receptor Cells Cultured chemistry.chemical_classification Membrane Glycoproteins Multidisciplinary Tight junction Chemistry Chemical Synthesis Brain Cell biology Signal Filtering Vesicular transport protein Transcytosis Blood-Brain Barrier Physical Sciences Melanotransferrin Medicine Engineering and Technology Cellular Structures and Organelles Cellular Types Anatomy Low Density Lipoprotein Receptor-Related Protein-1 Research Article Biosynthetic Techniques Science Materials Science Material Properties Transferrin receptor Research and Analysis Methods Permeability SDG 3 - Good Health and Well-being Receptors Transferrin Animals Humans Peptide Synthesis Fluorescent Dyes Biology and Life Sciences Endothelial Cells Epithelial Cells Cell Biology Bandpass Filters Biological Tissue Signal Processing Lysosomes Peptides |
| Zdroj: | Hudecz, D, Sigurdardóttir, S B, Christensen, S C, Hempel, C, Urquhart, A J, Andresen, T L & Nielsen, M S 2021, ' Two peptides targeting endothelial receptors are internalized into murine brain endothelial cells ', PLOS ONE, vol. 16, no. 4, e0249686 . https://doi.org/10.1371/journal.pone.0249686 Hudecz, D, Sigurdardóttir, S B, Christensen, S C, Hempel, C, Urquhart, A J, Andresen, T L & Nielsen, M S 2021, ' Two peptides targeting endothelial receptors are internalized into murine brain endothelial cells ', PLOS ONE, vol. 16, e0249686 . https://doi.org/10.1371/journal.pone.0249686 PLoS ONE PLoS ONE, Vol 16, Iss 4, p e0249686 (2021) |
| Popis: | The blood-brain barrier (BBB) is one of the main obstacles for therapies targeting brain diseases. Most macromolecules fail to pass the tight BBB, formed by brain endothelial cells interlinked by tight junctions. A wide range of small, lipid-soluble molecules can enter the brain parenchyma via diffusion, whereas macromolecules have to transcytose via vesicular transport. Vesicular transport can thus be utilized as a strategy to deliver brain therapies. By conjugating BBB targeting antibodies and peptides to therapeutic molecules or nanoparticles, it is possible to increase uptake into the brain. Previously, the synthetic peptide GYR and a peptide derived from melanotransferrin (MTfp) have been suggested as candidates for mediating transcytosis in brain endothelial cells (BECs). Here we study uptake, intracellular trafficking, and translocation of these two peptides in BECs. The peptides were synthesized, and binding studies to purified endocytic receptors were performed using surface plasmon resonance. Furthermore, the peptides were conjugated to a fluorophore allowing for live-cell imaging studies of their uptake into murine brain endothelial cells. Both peptides bound to low-density lipoprotein receptor-related protein 1 (LRP-1) and the human transferrin receptor, while lower affinity was observed against the murine transferrin receptor. The MTfp showed a higher binding affinity to all receptors when compared to the GYR peptide. The peptides were internalized by the bEnd.3 mouse endothelial cells within 30 min of incubation and frequently co-localized with endo-lysosomal vesicles. Moreover, our in vitro Transwell translocation experiments confirmed that GYR was able to cross the murine barrier and indicated the successful translocation of MTfp. Thus, despite binding to endocytic receptors with different affinities, both peptides are able to transcytose across the murine BECs. |
| Databáze: | OpenAIRE |
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