Since their discovery, liposomes have been widely employed in biomedical research. These nano-size spherical vesicles consisting one or few phospholipid bilayers surrounding an aqueous core are capable of carrying a wide variety of bioactive compounds, including drugs, peptides, nucleic acids, proteins and others. Despite considerable success achieved in synthesis of liposome constructs containing bioactive compounds, preparation of ligand-targeted liposomes comprising large quantities of encapsulated proteins that are capable of affecting pathological cells still remains a big challenge. Here we described a novel method for preparation of small (80–90 nm in diameter) unilamellar liposomes containing very large quantities (thousands of protein molecules per liposome) of heme-containing cytochrome c, highly fluorescent mCherry and highly toxic PE40 (Pseudomonas aeruginosa Exotoxin A domain). Efficient encapsulation of the proteins was achieved through electrostatic interaction between positively charged proteins (at pH lower than pI) and negatively charged liposome membrane. The proteoliposomes containing large quantities of mCherry or PE40 and functionalized with designed ankyrin repeat protein (DARPin)_9-29, which targets human epidermal growth factor receptor 2 (HER2) were shown to specifically stain and kill in sub-nanomolar concentrations HER2-positive cells, overexpressing HER2, respectively. Specific staining and eradication of the receptor-positive cells demonstrated here makes the DARPin-functionalized liposomes carrying large quantities of fluorescent and/or toxic proteins a promising candidate for tumor detection and therapy.

Selective staining and eradication of cancer cells by protein-carrying DARPin-functionalized liposomes / S. Deyev, G. Proshkina, O. Baryshnikova, A. Ryabova, G. Avishai, L. Katrivas, C. Giannini, Y. Levi-Kalisman, A. Kotlyar. - In: EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS. - ISSN 0939-6411. - 130(2018 Sep), pp. 296-305. [10.1016/j.ejpb.2018.06.026]

Selective staining and eradication of cancer cells by protein-carrying DARPin-functionalized liposomes

Giannini, Clelia;
2018-09

Abstract

Since their discovery, liposomes have been widely employed in biomedical research. These nano-size spherical vesicles consisting one or few phospholipid bilayers surrounding an aqueous core are capable of carrying a wide variety of bioactive compounds, including drugs, peptides, nucleic acids, proteins and others. Despite considerable success achieved in synthesis of liposome constructs containing bioactive compounds, preparation of ligand-targeted liposomes comprising large quantities of encapsulated proteins that are capable of affecting pathological cells still remains a big challenge. Here we described a novel method for preparation of small (80–90 nm in diameter) unilamellar liposomes containing very large quantities (thousands of protein molecules per liposome) of heme-containing cytochrome c, highly fluorescent mCherry and highly toxic PE40 (Pseudomonas aeruginosa Exotoxin A domain). Efficient encapsulation of the proteins was achieved through electrostatic interaction between positively charged proteins (at pH lower than pI) and negatively charged liposome membrane. The proteoliposomes containing large quantities of mCherry or PE40 and functionalized with designed ankyrin repeat protein (DARPin)_9-29, which targets human epidermal growth factor receptor 2 (HER2) were shown to specifically stain and kill in sub-nanomolar concentrations HER2-positive cells, overexpressing HER2, respectively. Specific staining and eradication of the receptor-positive cells demonstrated here makes the DARPin-functionalized liposomes carrying large quantities of fluorescent and/or toxic proteins a promising candidate for tumor detection and therapy.
Apoptosis; Cancer; Cryo-TEM; DARPin; Exotoxin A; Liposomes; mCherry; ADP Ribose Transferases; Adenocarcinoma; Animals; Ankyrin Repeat; Bacterial Toxins; Breast Neoplasms; CHO Cells; Cell Line, Tumor; Cricetinae; Cricetulus; Cytochromes c; Exotoxins; Female; Heme; Humans; Liposomes; Luminescent Proteins; Ovarian Neoplasms; Particle Size; Receptor, ErbB-2; Virulence Factors; Biotechnology
Settore BIO/10 - Biochimica
EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/599384
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