Enhanced Cellular Uptake of Albumin-Based Lyophilisomes when Functionalized with Cell-Penetrating Peptide TAT in HeLa Cells

Etienne van Bracht, Luuk R. M. Versteegden, Sarah Stolle, Wouter P. R. Verdurmen, Rob Woestenenk, Rene Raave, Theo Hafmans, Egbert Oosterwijk, Roland Brock, Toin H. van Kuppevelt, Willeke F. Daamen*

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

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    Abstract

    Lyophilisomes are a novel class of biodegradable proteinaceous nano/micrometer capsules with potential use as drug delivery carrier. Cell-penetrating peptides (CPPs) including the TAT peptide have been successfully implemented for intracellular delivery of a broad variety of cargos including various nanoparticulate pharmaceutical carriers. In the present study, lyophilisomes were modified using CPPs in order to achieve enhanced cellular uptake. Lyophilisomes were prepared by a freezing, annealing, and lyophilization method and a cystein-elongated TAT peptide was conjugated to the lyophilisomes using a heterobifunctional linker. Fluorescent-activated cell sorting (FACS) was utilized to acquire a lyophilisome population with a particle diameter smaller than 1000 nm. Cultured HeLa, OVCAR-3, Caco-2 and SKOV-3 cells were exposed to unmodified lyophilisomes and TAT-conjugated lyophilisomes and examined with FACS. HeLa cells were investigated in more detail using a trypan blue quenching assay, confocal microscopy, and transmission electron microscopy. TAT-conjugation strongly increased binding and cellular uptake of lyophilisomes in a time-dependent manner in vitro, as assessed by FACS. These results were confirmed by confocal microscopy. Transmission electron microscopy indicated rapid cellular uptake of TAT-conjugated lyophilisomes via phagocytosis and/or macropinocytosis. In conclusion, TAT-peptides conjugated to albumin-based lyophilisomes are able to enhance cellular uptake of lyophilisomes in HeLa cells.

    Original languageEnglish
    Article numbere110813
    Number of pages9
    JournalPLoS ONE
    Volume9
    Issue number11
    DOIs
    Publication statusPublished - 4-Nov-2014

    Keywords

    • DRUG-DELIVERY
    • CANCER THERAPEUTICS
    • CONTRAST AGENT
    • IN-VIVO
    • NANOPARTICLES
    • EFFICACY
    • PACLITAXEL
    • PROTEIN
    • INTERNALIZATION
    • CYTOTOXICITY

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