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Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins

Development of targeted treatment for colorectal cancer is crucial to avoid side effects. To harness the possibilities offered by microbiome engineering, we prepared safe multifunctional cancer cell‐targeting bacteria Lactococcus lactis. They displayed, on their surface, binding proteins for cancer‐...

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Autores principales: Plavec, Tina Vida, Mitrović, Ana, Perišić Nanut, Milica, Štrukelj, Borut, Kos, Janko, Berlec, Aleš
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449671/
https://www.ncbi.nlm.nih.gov/pubmed/34347360
http://dx.doi.org/10.1111/1751-7915.13907
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author Plavec, Tina Vida
Mitrović, Ana
Perišić Nanut, Milica
Štrukelj, Borut
Kos, Janko
Berlec, Aleš
author_facet Plavec, Tina Vida
Mitrović, Ana
Perišić Nanut, Milica
Štrukelj, Borut
Kos, Janko
Berlec, Aleš
author_sort Plavec, Tina Vida
collection PubMed
description Development of targeted treatment for colorectal cancer is crucial to avoid side effects. To harness the possibilities offered by microbiome engineering, we prepared safe multifunctional cancer cell‐targeting bacteria Lactococcus lactis. They displayed, on their surface, binding proteins for cancer‐associated transmembrane receptors epithelial cell adhesion molecule (EpCAM) and human epidermal growth factor receptor 2 (HER2) and co‐expressed an infrared fluorescent protein for imaging. Binding of engineered L. lactis to tumour antigens EpCAM and HER2 was confirmed and characterised in vitro using soluble receptors. The proof‐of‐principle of targeting was demonstrated on human cell lines HEK293, HT‐29 and Caco‐2 with fluorescent microscopy and flow cytometry. The highest L. lactis adhesion was seen for the HEK293 cells with the overexpressed tumour antigens, where colocalisation with their tumour antigens was seen for 39% and 67% of EpCAM‐targeting and HER2‐targeting bacteria, respectively. On the other hand, no binding was observed to HEK293 cells without tumour antigens, confirming the selectivity of the engineered L. lactis. Apart from cell targeting in static conditions, targeting ability of engineered L. lactis was also shown in conditions of constant flow of bacterial suspension over the HEK293 cells. Successful targeting by engineered L. lactis support the future use of these bacteria in biopharmaceutical delivery for the treatment of colorectal cancer.
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spelling pubmed-84496712021-09-24 Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins Plavec, Tina Vida Mitrović, Ana Perišić Nanut, Milica Štrukelj, Borut Kos, Janko Berlec, Aleš Microb Biotechnol Research Articles Development of targeted treatment for colorectal cancer is crucial to avoid side effects. To harness the possibilities offered by microbiome engineering, we prepared safe multifunctional cancer cell‐targeting bacteria Lactococcus lactis. They displayed, on their surface, binding proteins for cancer‐associated transmembrane receptors epithelial cell adhesion molecule (EpCAM) and human epidermal growth factor receptor 2 (HER2) and co‐expressed an infrared fluorescent protein for imaging. Binding of engineered L. lactis to tumour antigens EpCAM and HER2 was confirmed and characterised in vitro using soluble receptors. The proof‐of‐principle of targeting was demonstrated on human cell lines HEK293, HT‐29 and Caco‐2 with fluorescent microscopy and flow cytometry. The highest L. lactis adhesion was seen for the HEK293 cells with the overexpressed tumour antigens, where colocalisation with their tumour antigens was seen for 39% and 67% of EpCAM‐targeting and HER2‐targeting bacteria, respectively. On the other hand, no binding was observed to HEK293 cells without tumour antigens, confirming the selectivity of the engineered L. lactis. Apart from cell targeting in static conditions, targeting ability of engineered L. lactis was also shown in conditions of constant flow of bacterial suspension over the HEK293 cells. Successful targeting by engineered L. lactis support the future use of these bacteria in biopharmaceutical delivery for the treatment of colorectal cancer. John Wiley and Sons Inc. 2021-08-04 /pmc/articles/PMC8449671/ /pubmed/34347360 http://dx.doi.org/10.1111/1751-7915.13907 Text en © 2021 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley & Sons Ltd. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Plavec, Tina Vida
Mitrović, Ana
Perišić Nanut, Milica
Štrukelj, Borut
Kos, Janko
Berlec, Aleš
Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins
title Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins
title_full Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins
title_fullStr Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins
title_full_unstemmed Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins
title_short Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins
title_sort targeting of fluorescent lactococcus lactis to colorectal cancer cells through surface display of tumour‐antigen binding proteins
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449671/
https://www.ncbi.nlm.nih.gov/pubmed/34347360
http://dx.doi.org/10.1111/1751-7915.13907
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