Jacchetti E

Raimondi MT.

English

Archivio istituzionale della ricerca - Politecnico di Milano

Open camera or QR reader andscan code to access this articleand other resources online.TERMIS 2023 – European ChapterManchester Central Conference CentreManchester, UKMarch 28–31, 2023TISSUE ENGINEERING: Part AVolume 29, Numbers 13 and 14, 2023ª Mary Ann Liebert, Inc.DOI: 10.1089/ten.tea.2023.29043.abstractsDownloaded by LI CO SA 8180401 MI from www.liebertpub.com at 03/17/25. For personal use only.  OP-312 3D micro scaffolds as a frontier tool to model the EMT phenotype in cancer cells Emanuela Jacchetti, Manuela Teresa Raimondi Department of Chemistry, Materials and Chemical Engineering "G.Natta", Politecnico di Milano.   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)[1, 2]. Markers of EMT are loss of cellular contacts, β-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/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 conventional 3D 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, and β-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 spheroids influence nuclear β-catenin translocation. DISCUSSION & CONCLUSIONS: Our 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   Keywords: Microenvironment and niche engineering, In vitro microenvironments    Downloaded by LI CO SA 8180401 MI from www.liebertpub.com at 03/17/25. For personal use only. 

3D micro scaffolds as a frontier tool to model the EMT phenotype in cancer cells.

https://re.public.polimi.it/bitstream/11311/1285915/1/Jacchetti_termis_2023.pdf

3D micro scaffolds as a frontier tool to model the EMT phenotype in cancer cells.

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