PLA-PEG nanospheres decorated with phage display selected peptides as biomarkers for detection of human colorectal adeno
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PLA-PEG nanospheres decorated with phage display selected peptides as biomarkers for detection of human colorectal adenocarcinoma Aline de Souza 1 & William Castro-Borges 2 & Milton Hércules Guerra de Andrade 2 & Yara Cristina de Paiva Maia 3 & Luiz Ricardo Goulart 3 & Elisa Gomes Lanna 1 & Ana Carolina Ferreira de Brito 4 & Ana Paula Moreira Barboza 4 & Vanessa Carla Furtado Mosqueira 1 & Karina Taciana Santos Rubio 1
# Controlled Release Society 2020
Abstract Peptide-mediated targeting to colorectal cancer can increase selectivity and specificity of this cancer diagnosis acting as biomarkers. The present work aimed to select peptides using the phage display technique and associate the peptides with polymeric nanospheres in order to evaluate their cytotoxicity and selectivity during cell interaction with Caco-2 human colon tumor cell line. Two peptides identified by phage display (peptide-1 and peptide-2) were synthesized and exhibited purity higher than 84%. Poly(lactic acid)-block-polyethylene glycol nanospheres were prepared by nanoprecipitation and double emulsion methods in order to load the two peptides. Nanoparticles ranged in size from 114 to 150 nm and peptide encapsulation efficiency varied from 16 to 32%, depending on the methodology. No cytotoxic activity was observed towards Caco-2 tumor cell line, either free or loaded peptides in concentrations up to 3 μM at incubation times of 6 and 24 h, indicating safety as biomarkers. Fluorescein isothiocyanate–labeled peptides allowed evaluating selective interactions with Caco-2 cells, where peptide-1 entrapped in nanospheres showed greater intensity of co-localized cell fluorescence, in comparison to peptide-2. Peptide-1 loaded in nanospheres revealed promising to be investigated in further studies of selectivity with other human colon rectal cells as a potential biomarker. Keywords Colorectal cancer . Phage display . Peptides . Caco-2 . Nanoparticles . Biomarker . Image-guided diagnosis
Introduction Colorectal cancer (CRC) is the world’s fourth most deadly cancer. It represents a global health problem with an annual incidence reaching almost 900,000 deaths. Among these
* Vanessa Carla Furtado Mosqueira [email protected]; [email protected] 1
Laboratory of Pharmaceutics and Nanobiotechnology (LDGNano), School of Pharmacy, Universidade Federal de Ouro Preto, Minas Gerais, Brazil
2
Laboratory of Enzymology and Proteomics, NUPEB, Universidade Federal de Ouro Preto, Minas Gerais, Brazil
3
Laboratory of Nanobiotechnology, Institute of Biotechnology, Universidade Federal de Uberlandia, Minas Gerais, Brazil
4
Department of Physics, Exact and Biological Sciences Institute, Universidade Federal de Ouro Preto, Minas Gerais, Brazil
cases, the majority arises from adenomas, polyposis, genetic syndromes, or inflammatory bowel disease [1, 2]. Specific diagnosis is a global challenge. Currently, identification of occult blood in the stool is used as a screening test for CRC, although it lacks specificity [3]. Thereby, the use of biomarkers could help to i
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