Exosomes (EXOs) are considered an exceptionally promising source of cancer biomarkers for personalized medicine and liquid biopsy. Despite this potential, the EXOs translation process in diagnostics is still at its birth, and the development of reliable and reproducible methods for their characterization is highly demanded. Fourier Transform Infrared (FTIR) Spectroscopy is perfectly suited for this purpose, as it can provide a label-free biochemical profile of EXOs in terms of lipid, protein, and nucleic acid content. Here we evaluated the applicability of FTIR spectroscopy to the study of cancer-derived EXOs as a function of cell differentiation. For this purpose, we used N-acetyl-L-Cysteine (NAC) to induce a controlled differentiation in human colon carcinoma cells from a proliferative mesenchymal morphology to a less invasive epithelial phenotype, as measured with fluorescence and electron microscopy. EXOs derived from cells with different phenotypes showed significant variation in the relative intensity of the amide I-II and CH-stretching bands in the mid-IR range, indicating the spectroscopic lipid/protein ratio as an effective classification parameter. Additionally, we showed that different cell phenotypes are associated with a shape modification in these spectral bands that can be automatically detected by combining Principal Component Analysis (PCA) with Linear Discriminant Analysis (LDA). On the one hand, our study confirms that an FTIR analysis of EXOs allows scientists to precisely detect modifications occurring at the parental cell level; on the other hand, it unveils a set of effective spectral biomarkers able to monitoring cell changes from a mesenchymal to an epithelial phenotype, a clinically valuable piece of information considering that the epithelial-to-mesenchymal transition is a key step in the metastatic process.

Romano, S., Di Giacinto, F., Primiano, A., Gervasoni, J., Mazzini, A., Papi, M., Urbani, A., Serafino, A., De Spirito, M., Krasnowska, E. K., Ciasca, G., Label-free spectroscopic characterization of exosomes reveals cancer cell differentiation, <<ANALYTICA CHIMICA ACTA>>, 2022; 1192 (N/A): 339359-N/A. [doi:10.1016/j.aca.2021.339359N/A] [http://hdl.handle.net/10807/209422]

Label-free spectroscopic characterization of exosomes reveals cancer cell differentiation

Di Giacinto, F.
;
Primiano, A.;Gervasoni, J.;Papi, M.;Urbani, A.;De Spirito, M.
;
Ciasca, G.
2022

Abstract

Exosomes (EXOs) are considered an exceptionally promising source of cancer biomarkers for personalized medicine and liquid biopsy. Despite this potential, the EXOs translation process in diagnostics is still at its birth, and the development of reliable and reproducible methods for their characterization is highly demanded. Fourier Transform Infrared (FTIR) Spectroscopy is perfectly suited for this purpose, as it can provide a label-free biochemical profile of EXOs in terms of lipid, protein, and nucleic acid content. Here we evaluated the applicability of FTIR spectroscopy to the study of cancer-derived EXOs as a function of cell differentiation. For this purpose, we used N-acetyl-L-Cysteine (NAC) to induce a controlled differentiation in human colon carcinoma cells from a proliferative mesenchymal morphology to a less invasive epithelial phenotype, as measured with fluorescence and electron microscopy. EXOs derived from cells with different phenotypes showed significant variation in the relative intensity of the amide I-II and CH-stretching bands in the mid-IR range, indicating the spectroscopic lipid/protein ratio as an effective classification parameter. Additionally, we showed that different cell phenotypes are associated with a shape modification in these spectral bands that can be automatically detected by combining Principal Component Analysis (PCA) with Linear Discriminant Analysis (LDA). On the one hand, our study confirms that an FTIR analysis of EXOs allows scientists to precisely detect modifications occurring at the parental cell level; on the other hand, it unveils a set of effective spectral biomarkers able to monitoring cell changes from a mesenchymal to an epithelial phenotype, a clinically valuable piece of information considering that the epithelial-to-mesenchymal transition is a key step in the metastatic process.
Inglese
Romano, S., Di Giacinto, F., Primiano, A., Gervasoni, J., Mazzini, A., Papi, M., Urbani, A., Serafino, A., De Spirito, M., Krasnowska, E. K., Ciasca, G., Label-free spectroscopic characterization of exosomes reveals cancer cell differentiation, <<ANALYTICA CHIMICA ACTA>>, 2022; 1192 (N/A): 339359-N/A. [doi:10.1016/j.aca.2021.339359N/A] [http://hdl.handle.net/10807/209422]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10807/209422
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