Liver fibrosis (LF) is a major concern for public health worldwide, with more than 800 million people affected and a mortality rate of chronic liver diaseas of approximately 2 million deaths per year [1]. To date, there is still a great need to develop novel therapeutic strategies to treat LF. Moreover, liver fibrosis is one of the most well-known predisposing causes of hepatocellular carcinoma (HCC): clinical findings indicate that > 80% of HCCs develop in the contest of liver fibrosis/cirrhosis [2]. The main aim of this thesis was to study novel drugs with anti-fibrotic potential. To this end, we have used a class of deubiquitinases (DUBs) inhibitors named 2C and DUDC3, that we successfully employed to down regulating the growth of ovarian cancer cells [3]. To explore DUBs inhibitors effectiveness, we have considered three different experimental models: in the first where DUBs inhibitors effectiveness was studied in Hepatic Stellate Cells (HSCs – the main responsible of LF) cultivated in standard plastic dishes, in the second DUBs inhibitors were tested in HSCs grown in 3D structures (spheroids) and in the third DUBs inhibitors were evaluated in HSCs cultivated on surfaces able to mimic the normal and pathological liver viscoelastic properties. The first model employed, i.e. plastic dishes, does not resemble the real environment of the fibrotic liver for several reasons among which the fact plastic dishes exhibit significantly different mechanical properties than a cirrhotic liver [4]. In this regard, it is known that the increase of the stiffness and, therefore, the elastic properties promote cells activation and differentiation [5]. Despite this limitation, plastic dishes have the advantage of the easiness of cell culturing and of the further processing to study cell phenotype and molecular effects. The spheroid model has the advantage to explore the effects of the drugs in a 3D structure somehow resembling the structure of the liver, although with a simplified tissue architecture. Finally, the third model allows studying the effects of the drugs in cell cultivated on surfaces with realistic viscoelastic properties, i.e. those represented by the normal and the pathological fibrotic liver. This model is of particular relevance, as it is known that increased surface stiffness promotes cells activation and differentiation. As very few data about human liver viscoelastic properties are available in the literature, here we have determined them in samples obtained from normal and pathological liver. For this purpose, rheology technique was employed and the outcoming results were compared to those descending from the Low Field NMR characterization. The data obtained were used to prepare cell-culturing surface made by the polymer alginate, either with normal or pathological viscoelastic properties. Our data indicate that, in the three models employed, the drugs considered have the power to down regulating HSCs growth and fibrotic activation. While the three models have each some limitations, taken together they can provide data with a significant predictive power for studies performed in animal models of liver fibrosis. In this regard, the in vivo studies are ongoing in the lab of prof. Truong Hai Nhung (University of Sciences, Ho Chi Minh City, Vietnam) in the frame of the common project “A novel molecular approach to liver fibrosis” financed by the Ministry of Foreign Affairs and International Cooperation (MAECI) to Prof. Gabriele Grassi. References [1] Ye Y, et al., 2022. Front. Mol. Biosci. 9, 963630. [2] Affo S, et al., 2017. Annu. Rev. Pathol. Mech. Dis. 12, 153–186. [3] Maddaloni M, et al. 2024. Pharmaceutics 16, 664. [4] Mazza G, et al., 2017. Adv. Drug Deliv. Rev. 121, 147–157. [5] Olsen AL, et al., 2011. Am. J. Physiol.-Gastrointest. Liver Physiol. 301, G110–G118.
Liver fibrosis (LF) is a major concern for public health worldwide, with more than 800 million people affected and a mortality rate of chronic liver diaseas of approximately 2 million deaths per year [1]. To date, there is still a great need to develop novel therapeutic strategies to treat LF. Moreover, liver fibrosis is one of the most well-known predisposing causes of hepatocellular carcinoma (HCC): clinical findings indicate that > 80% of HCCs develop in the contest of liver fibrosis/cirrhosis [2]. The main aim of this thesis was to study novel drugs with anti-fibrotic potential. To this end, we have used a class of deubiquitinases (DUBs) inhibitors named 2C and DUDC3, that we successfully employed to down regulating the growth of ovarian cancer cells [3]. To explore DUBs inhibitors effectiveness, we have considered three different experimental models: in the first where DUBs inhibitors effectiveness was studied in Hepatic Stellate Cells (HSCs – the main responsible of LF) cultivated in standard plastic dishes, in the second DUBs inhibitors were tested in HSCs grown in 3D structures (spheroids) and in the third DUBs inhibitors were evaluated in HSCs cultivated on surfaces able to mimic the normal and pathological liver viscoelastic properties. The first model employed, i.e. plastic dishes, does not resemble the real environment of the fibrotic liver for several reasons among which the fact plastic dishes exhibit significantly different mechanical properties than a cirrhotic liver [4]. In this regard, it is known that the increase of the stiffness and, therefore, the elastic properties promote cells activation and differentiation [5]. Despite this limitation, plastic dishes have the advantage of the easiness of cell culturing and of the further processing to study cell phenotype and molecular effects. The spheroid model has the advantage to explore the effects of the drugs in a 3D structure somehow resembling the structure of the liver, although with a simplified tissue architecture. Finally, the third model allows studying the effects of the drugs in cell cultivated on surfaces with realistic viscoelastic properties, i.e. those represented by the normal and the pathological fibrotic liver. This model is of particular relevance, as it is known that increased surface stiffness promotes cells activation and differentiation. As very few data about human liver viscoelastic properties are available in the literature, here we have determined them in samples obtained from normal and pathological liver. For this purpose, rheology technique was employed and the outcoming results were compared to those descending from the Low Field NMR characterization. The data obtained were used to prepare cell-culturing surface made by the polymer alginate, either with normal or pathological viscoelastic properties. Our data indicate that, in the three models employed, the drugs considered have the power to down regulating HSCs growth and fibrotic activation. While the three models have each some limitations, taken together they can provide data with a significant predictive power for studies performed in animal models of liver fibrosis. In this regard, the in vivo studies are ongoing in the lab of prof. Truong Hai Nhung (University of Sciences, Ho Chi Minh City, Vietnam) in the frame of the common project “A novel molecular approach to liver fibrosis” financed by the Ministry of Foreign Affairs and International Cooperation (MAECI) to Prof. Gabriele Grassi. References [1] Ye Y, et al., 2022. Front. Mol. Biosci. 9, 963630. [2] Affo S, et al., 2017. Annu. Rev. Pathol. Mech. Dis. 12, 153–186. [3] Maddaloni M, et al. 2024. Pharmaceutics 16, 664. [4] Mazza G, et al., 2017. Adv. Drug Deliv. Rev. 121, 147–157. [5] Olsen AL, et al., 2011. Am. J. Physiol.-Gastrointest. Liver Physiol. 301, G110–G118.
Rheological and Low Field NMR properties of hepatic tissue and polymeric gels in the study of deubiquitinase inhibitors against liver fibrosis / Biasin, Alice. - (2025 Jan 27).
Rheological and Low Field NMR properties of hepatic tissue and polymeric gels in the study of deubiquitinase inhibitors against liver fibrosis
BIASIN, ALICE
2025-01-27
Abstract
Liver fibrosis (LF) is a major concern for public health worldwide, with more than 800 million people affected and a mortality rate of chronic liver diaseas of approximately 2 million deaths per year [1]. To date, there is still a great need to develop novel therapeutic strategies to treat LF. Moreover, liver fibrosis is one of the most well-known predisposing causes of hepatocellular carcinoma (HCC): clinical findings indicate that > 80% of HCCs develop in the contest of liver fibrosis/cirrhosis [2]. The main aim of this thesis was to study novel drugs with anti-fibrotic potential. To this end, we have used a class of deubiquitinases (DUBs) inhibitors named 2C and DUDC3, that we successfully employed to down regulating the growth of ovarian cancer cells [3]. To explore DUBs inhibitors effectiveness, we have considered three different experimental models: in the first where DUBs inhibitors effectiveness was studied in Hepatic Stellate Cells (HSCs – the main responsible of LF) cultivated in standard plastic dishes, in the second DUBs inhibitors were tested in HSCs grown in 3D structures (spheroids) and in the third DUBs inhibitors were evaluated in HSCs cultivated on surfaces able to mimic the normal and pathological liver viscoelastic properties. The first model employed, i.e. plastic dishes, does not resemble the real environment of the fibrotic liver for several reasons among which the fact plastic dishes exhibit significantly different mechanical properties than a cirrhotic liver [4]. In this regard, it is known that the increase of the stiffness and, therefore, the elastic properties promote cells activation and differentiation [5]. Despite this limitation, plastic dishes have the advantage of the easiness of cell culturing and of the further processing to study cell phenotype and molecular effects. The spheroid model has the advantage to explore the effects of the drugs in a 3D structure somehow resembling the structure of the liver, although with a simplified tissue architecture. Finally, the third model allows studying the effects of the drugs in cell cultivated on surfaces with realistic viscoelastic properties, i.e. those represented by the normal and the pathological fibrotic liver. This model is of particular relevance, as it is known that increased surface stiffness promotes cells activation and differentiation. As very few data about human liver viscoelastic properties are available in the literature, here we have determined them in samples obtained from normal and pathological liver. For this purpose, rheology technique was employed and the outcoming results were compared to those descending from the Low Field NMR characterization. The data obtained were used to prepare cell-culturing surface made by the polymer alginate, either with normal or pathological viscoelastic properties. Our data indicate that, in the three models employed, the drugs considered have the power to down regulating HSCs growth and fibrotic activation. While the three models have each some limitations, taken together they can provide data with a significant predictive power for studies performed in animal models of liver fibrosis. In this regard, the in vivo studies are ongoing in the lab of prof. Truong Hai Nhung (University of Sciences, Ho Chi Minh City, Vietnam) in the frame of the common project “A novel molecular approach to liver fibrosis” financed by the Ministry of Foreign Affairs and International Cooperation (MAECI) to Prof. Gabriele Grassi. References [1] Ye Y, et al., 2022. Front. Mol. Biosci. 9, 963630. [2] Affo S, et al., 2017. Annu. Rev. Pathol. Mech. Dis. 12, 153–186. [3] Maddaloni M, et al. 2024. Pharmaceutics 16, 664. [4] Mazza G, et al., 2017. Adv. Drug Deliv. Rev. 121, 147–157. [5] Olsen AL, et al., 2011. Am. J. Physiol.-Gastrointest. Liver Physiol. 301, G110–G118.Pubblicazioni consigliate
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