Endosomal escape of protein nanoparticles engineered through humanized histidine-rich peptides  

作　　者：Hector Lopez-Laguna Rafael Cubarsi Ugutz Unzueta Ramon Mangues Esther Vazquez Antonio Villaverde 

机构地区：[1]Institut de Biotecnologia i de Biomedicina,Universitat Aut6noma de Barcelona,Bellaterra,08193 Barcelona,Spain [2]Departament de Genetica i de Microbiologia,Universitat Aut6noma de Barcelona,Bellaterra,08193 Barcelona,Spain [3]CIBER de Bioingenieria,Biomateriales y Nanomedicina(CIBER-BBN),C/Monforte de Lemos 3-5,28029 Madrid,Spain [4]Departament de Matematiques,Universitat Politecnica de Catalunya,08034,Barcelona,Spain [5]Institut d'Investigacions Biomediques Sant Pau,Hospital de la Santa Creu i Sant Pau,08025 Barcelona,Spain

出　　处：《Science China Materials》2020年第4期644-653,共10页中国科学（材料科学（英文版）

基　　金：Agencia Estatal de Investigación (AEI) and to Fondo Europeo de Desarrollo Regional (FEDER) (BIO2016-76063-R, AEI/FEDER, UE) to Villaverde A, AGAUR (2017SGR-229) to Villaverde A and 2017SGR-865 GRC;ISCⅢ (PI15/00272 co-founding FEDER) to Vázquez E and ISCⅢ (Co-founding FEDER) PIE15//00028 and PI18/00650 to Mangues R, and to EU COST Action CA 17140;funded by the Ⅵ National R&D&I Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions;financed by the Instituto de Salud Carlos Ⅲ, with assistance from the European Regional Development Fund;supported by a predoctoral fellowship from AGAUR (2019 FI_B 00352);PERIS program from the Health Department of the Generalitat de Catalunya

摘　　要：Poly-histidine peptides such as H6(HHHHHH)are used in protein biotechnologies as purification tags,protein-assembling agents and endosomal-escape entities.The pleiotropic properties of such peptides make them appealing to design protein-based smart materials or nanoparticles for imaging or drug delivery to be produced in form of recombinant proteins.However,the clinical applicability of H6-tagged proteins is restricted by the potential immunogenicity of these segments.In this study,we have explored several humanized histidine-rich peptides in tumor-targeted modular proteins,which can specifically bind and be internalized by the target cells through the tumoral marker CXCR4.We were particularly interested in exploring how protein purification,self-assembling and endosomal escape perform in proteins containing the variant histidine-rich tags.Among the tested candidates,the peptide H5 E(HEHEHEHEH)is promising as a good promoter of endosomal escape of the associated fulllength protein upon endosomal internalization.The numerical modelling of cell penetration and endosomal escape of the tested proteins has revealed a negative relationship between the amount of protein internalized into target cells and the efficiency of cytoplasmic release.This fact demonstrates that the His-mediated,proton sponge-based endosomal escape saturates at moderate amounts of internalized protein,a fact that might be critical for the design of protein materials for cytosolic molecular delivery.Poly-histidine peptides such as H6(HHHHHH)are used in protein biotechnologies as purification tags, protein-assembling agents and endosomal-escape entities. The pleiotropic properties of such peptides make them appealing to design protein-based smart materials or nanoparticles for imaging or drug delivery to be produced in form of recombinant proteins. However, the clinical applicability of H6-tagged proteins is restricted by the potential immunogenicity of these segments. In this study, we have explored several humanized histidine-rich peptides in tumor-targeted modular proteins, which can specifically bind and be internalized by the target cells through the tumoral marker CXCR4. We were particularly interested in exploring how protein purification,self-assembling and endosomal escape perform in proteins containing the variant histidine-rich tags. Among the tested candidates, the peptide H5 E(HEHEHEHEH) is promising as a good promoter of endosomal escape of the associated fulllength protein upon endosomal internalization. The numerical modelling of cell penetration and endosomal escape of the tested proteins has revealed a negative relationship between the amount of protein internalized into target cells and the efficiency of cytoplasmic release. This fact demonstrates that the His-mediated, proton sponge-based endosomal escape saturates at moderate amounts of internalized protein, a fact that might be critical for the design of protein materials for cytosolic molecular delivery.

关 键 词：protein materials NANOPARTICLES genetic design endosomal escape poly-histidines 

分 类 号：R318.08[医药卫生—生物医学工程]