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Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo

Urinary bladder cancer can be treated by intravesical application of therapeutic agents, but the specific targeting of cancer urothelial cells and the endocytotic pathways of the agents are not known. During carcinogenesis, the superficial urothelial cells exhibit changes in sugar residues on the ap...

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Autores principales: Resnik, Nataša, Višnjar, Tanja, Smrkolj, Tomaž, Kreft, Mateja Erdani, Romih, Rok, Zupančič, Daša
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10624759/
https://www.ncbi.nlm.nih.gov/pubmed/37535087
http://dx.doi.org/10.1007/s00418-023-02224-2
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author Resnik, Nataša
Višnjar, Tanja
Smrkolj, Tomaž
Kreft, Mateja Erdani
Romih, Rok
Zupančič, Daša
author_facet Resnik, Nataša
Višnjar, Tanja
Smrkolj, Tomaž
Kreft, Mateja Erdani
Romih, Rok
Zupančič, Daša
author_sort Resnik, Nataša
collection PubMed
description Urinary bladder cancer can be treated by intravesical application of therapeutic agents, but the specific targeting of cancer urothelial cells and the endocytotic pathways of the agents are not known. During carcinogenesis, the superficial urothelial cells exhibit changes in sugar residues on the apical plasma membranes. This can be exploited for selective targeting from the luminal side of the bladder. Here we show that the plant lectins Jacalin (from Artocarpus integrifolia), ACA (from Amaranthus caudatus) and DSA (from Datura stramonium) selectively bind to the apical plasma membrane of low- (RT4) and high-grade (T24) cancer urothelial cells in vitro and urothelial tumours ex vivo. The amount of lectin binding was significantly different between RT4 and T24 cells. Endocytosis of lectins was observed only in cancer urothelial cells and not in normal urothelial cells. Transmission electron microscopy analysis showed macropinosomes, endosome-like vesicles and multivesicular bodies filled with lectins in RT4 and T24 cells and also in cells of urothelial tumours ex vivo. Endocytosis of Jacalin and ACA in cancer cells was decreased in vitro after addition of inhibitor of macropinocytosis 5-(N-ethyl-N-isopropyl) amiloride (EIPA) and increased after stimulation of macropinocytosis with epidermal growth factor (EGF). Clathrin, caveolin and flotillin did not colocalise with lectins. These results confirm that the predominant mechanism of lectin endocytosis in cancer urothelial cells is macropinocytosis. Therefore, we propose that lectins in combination with conjugated therapeutic agents are promising tools for improved intravesical therapy by targeting cancer cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00418-023-02224-2.
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spelling pubmed-106247592023-11-05 Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo Resnik, Nataša Višnjar, Tanja Smrkolj, Tomaž Kreft, Mateja Erdani Romih, Rok Zupančič, Daša Histochem Cell Biol Original Paper Urinary bladder cancer can be treated by intravesical application of therapeutic agents, but the specific targeting of cancer urothelial cells and the endocytotic pathways of the agents are not known. During carcinogenesis, the superficial urothelial cells exhibit changes in sugar residues on the apical plasma membranes. This can be exploited for selective targeting from the luminal side of the bladder. Here we show that the plant lectins Jacalin (from Artocarpus integrifolia), ACA (from Amaranthus caudatus) and DSA (from Datura stramonium) selectively bind to the apical plasma membrane of low- (RT4) and high-grade (T24) cancer urothelial cells in vitro and urothelial tumours ex vivo. The amount of lectin binding was significantly different between RT4 and T24 cells. Endocytosis of lectins was observed only in cancer urothelial cells and not in normal urothelial cells. Transmission electron microscopy analysis showed macropinosomes, endosome-like vesicles and multivesicular bodies filled with lectins in RT4 and T24 cells and also in cells of urothelial tumours ex vivo. Endocytosis of Jacalin and ACA in cancer cells was decreased in vitro after addition of inhibitor of macropinocytosis 5-(N-ethyl-N-isopropyl) amiloride (EIPA) and increased after stimulation of macropinocytosis with epidermal growth factor (EGF). Clathrin, caveolin and flotillin did not colocalise with lectins. These results confirm that the predominant mechanism of lectin endocytosis in cancer urothelial cells is macropinocytosis. Therefore, we propose that lectins in combination with conjugated therapeutic agents are promising tools for improved intravesical therapy by targeting cancer cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00418-023-02224-2. Springer Berlin Heidelberg 2023-08-03 2023 /pmc/articles/PMC10624759/ /pubmed/37535087 http://dx.doi.org/10.1007/s00418-023-02224-2 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Original Paper
Resnik, Nataša
Višnjar, Tanja
Smrkolj, Tomaž
Kreft, Mateja Erdani
Romih, Rok
Zupančič, Daša
Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo
title Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo
title_full Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo
title_fullStr Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo
title_full_unstemmed Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo
title_short Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo
title_sort selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10624759/
https://www.ncbi.nlm.nih.gov/pubmed/37535087
http://dx.doi.org/10.1007/s00418-023-02224-2
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