E. Jacchetti

G. Cerullo

M. T. Raimondi

R. Osellame

English

Archivio istituzionale della ricerca - Politecnico di Milano

 Nanoengineering for Mechanobiology – n4m.mechanobiology.eu N4MNanoengineering for Mechanobiology 2024 Mechanobiology of Molecular  and Cellular Systems  3-7 March 2024 Camogli, Genova, Italy  Abstracts    Sponsors and Supporters                 Nanoengineering for Mechanobiology 2024 – n4m.mechanobiology.eu-2024 N4M Epithelial to Mesenchymal Transition modulation in breast cancer cells: a frontier for new investigation approaches   Manuela Raimondi1*, Giulio Cerullo2, Roberto Osellame2, Emanuela Jacchetti1* 1 Department of Chemistry, Materials and Chemical Engineering "G.Natta" Politecnico di Milano. 2Istituto di Fotonica e Nanotecnologie (IFN)-CNR and Department of Physics, Politecnico di Milano, 20133 Milan, Italy; emanuela.jacchetti@polimi.it Headline Over 90% of cancer deaths are due to metastases, however the effectiveness of therapy in reducing them has reached a plateau. To increase our arsenal for one more effective treatment, we need new targets and therefore our knowledge and the investigation approaches. Here, we analyse the ability of a 3D photopolymerized substrate to investigate breast cancer mechanotransduction in EMT. Abstract INTRODUCTION  Breast cancer is the most frequently diagnosed cancer, and the leading cause of death, among women. Its onset is associated with phenotype transition from epithelial to mesenchymal (EMT), a process generating cancer stem cells (CSCs) -catenin nuclear localization, cytoskeleton reorganisation, vimentin expression and increased motility [3]. There are several factors of the tumour microenvironment known to influence EMT, such as hypoxia, immune response and stiffening of the extracellular matrix [4]. We developed a culture substrate, called Niche, which is a custom-made, microporous, optically accessible 3D scaffold fabricated by 2-photon laser polymerization [5], in which cells adhere and grow in a spatial configuration different from the spheroid standard culture condition. Here, we analyse the ability of this substrate to sustain the EMT phenotype in CSCs. METHODS Malignant breast cancer cell lines (MCF7 and MDA-MB-232) were grown in different cell culture conditions: on 2D flat glass coverslips, on 2D soft collagen based-hydrogels (with Young’s modulus of 10Pa), in the 3D Niche, and in conventional3D spheroids. We used a pharmacological control to destroy actin cytoskeleton and reduce intracellular tension. Immunofluorescence assays were used to characterize the organization and localization of EMT-key markers: actin, vimentin, -catenin. The role of the various substrates in modulating cell migration was investigated by time-lapse microscopy. RESULTS Results show that the Niche and the pharmacological control limit the cell capability to migrate. The actin fibers number, related to intracellular tension, is highest on flat, and reduced in the Niche and spheroids. Vimentin is less expressed in the Niche than 2D glass and protein localization in the Niche is similar to spheroid samples. Only the Niche and the -catenin translocation. DISCUSSION & CONCLUSIONS Nanoengineering for Mechanobiology 2024 – n4m.mechanobiology.eu-2024 N4MOur results suggest that the Niche, influencing cellular adhesion properties, can modify key parameters related to cancer phenotype, primarily cell motility. This substrate also guarantees optical access to the samples to monitor in real-time the phenomenon evolution. Therefore, the Niche is a valuable tool for 3D-cancer-models and pharmacological research in vitro. We are now using this substrate to evaluate how mechanobiology can improve knowledge and research approaches related to cancer biology. ACKNOWLEDGEMENTS: ERC-project BEACONSANDEGG G.A.10105312 References [1] Abraham et al. Clin Cancer Res. 2005 11(3):1154-9 [2] Sheridan et al. Breast Cancer Res. 2006;8(5):R59 [3] Singh et al. Pharmacol Ther. 2018 182:80-94 [4] Gonzalez et al. Sci Signal. 2014 7(344):re8 [5] Jacchetti et al. Sci Rep. 2021 Feb 4;11(1):3021  Figure 1. Mechanotransduction investigation of EMT: concept of the work  

Epithelial to Mesenchymal Transition modulation in breast cancer cells: a frontier for new investigation approaches.

https://re.public.polimi.it/bitstream/11311/1285987/1/Jacchetti_N4M_2024.pdf

Epithelial to Mesenchymal Transition modulation in breast cancer cells: a frontier for new investigation approaches.

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