Intracellular delivery of functionally active proteins using self-assembling pyridylthiourea-polyethylenimine

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• 作者：Viktoriia Postupalenko^a ; Annie-Paule Sibler^b ; Dominique Desplancq^b ; Yves Nominé ; ^b ; Daniè ; le Spehner^c ; Patrick Schultz^c ; Etienne Weiss^b ; Guy Zuber^a ; ^{zuber@unistra.fr" class="auth_mail}
• 关键词：Intracellular protein delivery ; Antibodies ; Polymer ; Delivery vehicle ; Polyethylenimine ; Transfection
• 刊名：Journal of Controlled Release
• 出版年：28 March, 2014
• 年：2014
• 卷：178
• 期：Complete
• 页码：86-94
• 全文大小：1586 K

文摘

Intracellular delivery of functionally active proteins into cells is emerging as a novel strategy for research and therapeutic applications. Here, we present the properties of a self-assembling pyridylthiourea-modified polyethylenimine (蟺PEI), which interacts with proteins and promotes their delivery into the cytosol of mammalian cells. In aqueous medium at pH 7.4, self-association of 蟺PEI in the presence of green fluorescent proteins (GFP) leads to supramolecular protein-entrapped assemblies. These assemblies protect GFP from losing its fluorescence upon pH variation and assist delivery/translocation into the cytosol of mammalian cells via the endocytic pathway. The scope of application of this delivery system was extended to antibodies against intracellular targets as illustrated using a monoclonal antibody directed against the HPV-16 viral E6 oncoprotein and an antibody directed against the threonine-927 phosporylation site of the EG5 kinesin spindle protein. The 蟺PEI-mediated delivery of native anti-E6 antibodies or anti-E6 antibodies equipped with a nuclear localization signal (NLS), led to regeneration of the p53 tumor suppression protein in E6-transformed CaSki cells. Delivery of functionally active anti-EG5 antibodies, with the same polymer, reduced HeLa cell viability and appeared to perturb, as expected, chromosome segregation during mitosis. Altogether, these results provide an easy to use delivery system for extending the scope of application of antibodies for epitope recognition within living cells and may provide novel opportunities for selective interference of cell function by a steric hindrance modality.