Breast cancer remains a primary health concern among women worldwide, and insight into its initiation, progression, and metastasis are of the utmost importance. The role of Extracellular Vesicles (EVs), cell-derived membrane- surrounded vesicles, in cell-to-cell communication has long been recognized as a crucial component in these processes. A wide range of studies based on EVs are carried out on in vitro models, based on bi-dimensional (2D) cultures; these studies led to a greater understanding not only of the different mechanisms by which EVs contribute to tumor progression but also of their potential clinical use in diagnostics as biomarkers. Thus, investigating the molecular composition of EVs is of considerable importance, since it reflects the molecular composition of the parental cells [1]. Among all the techniques already set up, Fourier Transform Infrared spectroscopy (FTIR) has recently been introduced to the study of EVs. This innovative approach, commonly used in the material sciences field, enables fast label-free profiling, obtaining a distinctive molecular ‘fingerprint’ of EVs from minimal sample quantities, allowing the discovery of potential differences in molecular composition [2]. For this reason, for our study, EVs were isolated from two breast cancer cell lines, with different aggressiveness levels, cultured in 2D; EVs were isolated by ultracentrifugation technique and, once resuspended in a saline solution, they were analyzed for their FTIR spectra, to evaluate whether there could be differences that could reflect the different tumor grades. The results showed that there are some differences in several spectral regions of the two EVs samples, suggesting that the FTIR could allow the recognition of EVs, based on cell origin. However, the 2D models aren’t able to truthfully mimic the true complexity of the in vivo tumor microenvironment and the interactions mediated by EVs in it. Thus, more recent studies concerning EVs were focused on in vitro three-dimensional (3D) cell cultures, using tumor spheroids models [3]. To evaluate if there could be a difference in the molecular fingerprint of EVs derived from cells cultured in 2D or 3D, we isolated the EVs from breast tumor cells cultured by these two models and FTIR was performed on them. The results showed that the EVs isolated from 3D culture have different content in some molecular components compared to the EVs isolated from the respective cell lines cultured on 2D, suggesting that the composition of EVs is profoundly influenced by the cell culture methods and leading us to reflect on a careful evaluation of the most suitable cellular models in the in vitro studies. In conclusion, our preliminary data suggest that the FTIR offers new directions for EVs molecular fingerprints, making this tool potentially useful for biomarker discovery in liquid biopsy based on EVs.
Unleash the potential of Fourier-Transform Infrared (FT-IR) spectroscopy to study extracellular vesicles cancer
Poppa, Giuseppina;di Fazio, Giulia;D’Ascenzo, Sandra;Dolo, Vincenza;Giusti Ilaria.
2024-01-01
Abstract
Breast cancer remains a primary health concern among women worldwide, and insight into its initiation, progression, and metastasis are of the utmost importance. The role of Extracellular Vesicles (EVs), cell-derived membrane- surrounded vesicles, in cell-to-cell communication has long been recognized as a crucial component in these processes. A wide range of studies based on EVs are carried out on in vitro models, based on bi-dimensional (2D) cultures; these studies led to a greater understanding not only of the different mechanisms by which EVs contribute to tumor progression but also of their potential clinical use in diagnostics as biomarkers. Thus, investigating the molecular composition of EVs is of considerable importance, since it reflects the molecular composition of the parental cells [1]. Among all the techniques already set up, Fourier Transform Infrared spectroscopy (FTIR) has recently been introduced to the study of EVs. This innovative approach, commonly used in the material sciences field, enables fast label-free profiling, obtaining a distinctive molecular ‘fingerprint’ of EVs from minimal sample quantities, allowing the discovery of potential differences in molecular composition [2]. For this reason, for our study, EVs were isolated from two breast cancer cell lines, with different aggressiveness levels, cultured in 2D; EVs were isolated by ultracentrifugation technique and, once resuspended in a saline solution, they were analyzed for their FTIR spectra, to evaluate whether there could be differences that could reflect the different tumor grades. The results showed that there are some differences in several spectral regions of the two EVs samples, suggesting that the FTIR could allow the recognition of EVs, based on cell origin. However, the 2D models aren’t able to truthfully mimic the true complexity of the in vivo tumor microenvironment and the interactions mediated by EVs in it. Thus, more recent studies concerning EVs were focused on in vitro three-dimensional (3D) cell cultures, using tumor spheroids models [3]. To evaluate if there could be a difference in the molecular fingerprint of EVs derived from cells cultured in 2D or 3D, we isolated the EVs from breast tumor cells cultured by these two models and FTIR was performed on them. The results showed that the EVs isolated from 3D culture have different content in some molecular components compared to the EVs isolated from the respective cell lines cultured on 2D, suggesting that the composition of EVs is profoundly influenced by the cell culture methods and leading us to reflect on a careful evaluation of the most suitable cellular models in the in vitro studies. In conclusion, our preliminary data suggest that the FTIR offers new directions for EVs molecular fingerprints, making this tool potentially useful for biomarker discovery in liquid biopsy based on EVs.Pubblicazioni consigliate
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