Fibrosis is a complex scarring process that impairs tissue function, particularly concerning cardiac fibrosis, which affects the heart and can lead to heart dysfunction. Myofibroblasts play a central role in driving cardiac fibrosis, and epigenetic mechanisms may regulate their transformation. Epigenetic changes, such as DNA methylation and non-coding RNA activity, can lock cells into a pro-fibrotic state, perpetuating fibrosis. Also, chronic hypoxia, characterized by low tissue oxygen levels, contributes to the maintenance of this type of disease. Of note, diabetes, specifically type 2 diabetes (T2DM), which involves elevated blood glucose levels, is linked to epigenetic changes leading to an altered gene expression, promoting among other complications, cardiac fibrosis. Recent findings underline the important role of epigenetic processes, in particular DNA methylation, in the development of both diabetes and fibrosis. This modification is governed by a complex network of enzymes, including DNA methyltransferases (DNMTs) and Ten-eleven translocation proteins (TETs), whose activity depends on cofactors produced during fundamental metabolic processes. This thesis aims to delving into the complex interaction among DNA methylation, hypoxia, fibrosis, and diabetes, with a specific emphasis on the discovery of innovative drugs that possess epigenetic activity and hold the potential to alleviate these processes. A library comprising ten non-nucleoside analogue compounds, selectively inhibiting TET2, has undergone testing for their epigenetic and anti-fibrotic effects. Among these compounds, at least four demonstrated a significant increase in global DNA methylation in human cardiac fibroblasts under hypoxic conditions. Additionally, two of these compounds exhibited a notable reduction in the expression of fibrosis-related genes and proteins when compared to a solvent control. These two compounds also restored gene and protein levels to those observed under normoxic conditions. Cardiac fibroblasts, given their pivotal role in fibrosis research, are a primary focus, as inhibiting fibroblast-mediated ECM synthesis stands as a principal objective in antifibrotic therapeutic strategies. At first only foetal cardiac fibroblasts were taken into consideration, then the study developed comparing normoglycaemic versus hyperglycaemic patients derived cells. The results of this research provide evidence for a novel class of TET2-selective non-nucleoside analogues capable of modulating DNA methylation. These compounds hold the potential to become valuable tools in the prevention and reduction of cardiac fibrosis.
Progettazione e caratterizzazione di farmaci epigenetici per prevenire o trattare la fibrosi in modelli cellulari umani. Fibrosis is a complex scarring process that impairs tissue function, particularly concerning cardiac fibrosis, which affects the heart and can lead to heart dysfunction. Myofibroblasts play a central role in driving cardiac fibrosis, and epigenetic mechanisms may regulate their transformation. Epigenetic changes, such as DNA methylation and non-coding RNA activity, can lock cells into a pro-fibrotic state, perpetuating fibrosis. Also, chronic hypoxia, characterized by low tissue oxygen levels, contributes to the maintenance of this type of disease. Of note, diabetes, specifically type 2 diabetes (T2DM), which involves elevated blood glucose levels, is linked to epigenetic changes leading to an altered gene expression, promoting among other complications, cardiac fibrosis. Recent findings underline the important role of epigenetic processes, in particular DNA methylation, in the development of both diabetes and fibrosis. This modification is governed by a complex network of enzymes, including DNA methyltransferases (DNMTs) and Ten-eleven translocation proteins (TETs), whose activity depends on cofactors produced during fundamental metabolic processes. This thesis aims to delving into the complex interaction among DNA methylation, hypoxia, fibrosis, and diabetes, with a specific emphasis on the discovery of innovative drugs that possess epigenetic activity and hold the potential to alleviate these processes. A library comprising ten non-nucleoside analogue compounds, selectively inhibiting TET2, has undergone testing for their epigenetic and anti-fibrotic effects. Among these compounds, at least four demonstrated a significant increase in global DNA methylation in human cardiac fibroblasts under hypoxic conditions. Additionally, two of these compounds exhibited a notable reduction in the expression of fibrosis-related genes and proteins when compared to a solvent control. These two compounds also restored gene and protein levels to those observed under normoxic conditions. Cardiac fibroblasts, given their pivotal role in fibrosis research, are a primary focus, as inhibiting fibroblast-mediated ECM synthesis stands as a principal objective in antifibrotic therapeutic strategies. At first only foetal cardiac fibroblasts were taken into consideration, then the study developed comparing normoglycaemic versus hyperglycaemic patients derived cells. The results of this research provide evidence for a novel class of TET2-selective non-nucleoside analogues capable of modulating DNA methylation. These compounds hold the potential to become valuable tools in the prevention and reduction of cardiac fibrosis.
Epigenetic drug design and characterization to prevent or ameliorate fibrosis in human cellular systems
GAVAZZO, GAIA
2022/2023
Abstract
Fibrosis is a complex scarring process that impairs tissue function, particularly concerning cardiac fibrosis, which affects the heart and can lead to heart dysfunction. Myofibroblasts play a central role in driving cardiac fibrosis, and epigenetic mechanisms may regulate their transformation. Epigenetic changes, such as DNA methylation and non-coding RNA activity, can lock cells into a pro-fibrotic state, perpetuating fibrosis. Also, chronic hypoxia, characterized by low tissue oxygen levels, contributes to the maintenance of this type of disease. Of note, diabetes, specifically type 2 diabetes (T2DM), which involves elevated blood glucose levels, is linked to epigenetic changes leading to an altered gene expression, promoting among other complications, cardiac fibrosis. Recent findings underline the important role of epigenetic processes, in particular DNA methylation, in the development of both diabetes and fibrosis. This modification is governed by a complex network of enzymes, including DNA methyltransferases (DNMTs) and Ten-eleven translocation proteins (TETs), whose activity depends on cofactors produced during fundamental metabolic processes. This thesis aims to delving into the complex interaction among DNA methylation, hypoxia, fibrosis, and diabetes, with a specific emphasis on the discovery of innovative drugs that possess epigenetic activity and hold the potential to alleviate these processes. A library comprising ten non-nucleoside analogue compounds, selectively inhibiting TET2, has undergone testing for their epigenetic and anti-fibrotic effects. Among these compounds, at least four demonstrated a significant increase in global DNA methylation in human cardiac fibroblasts under hypoxic conditions. Additionally, two of these compounds exhibited a notable reduction in the expression of fibrosis-related genes and proteins when compared to a solvent control. These two compounds also restored gene and protein levels to those observed under normoxic conditions. Cardiac fibroblasts, given their pivotal role in fibrosis research, are a primary focus, as inhibiting fibroblast-mediated ECM synthesis stands as a principal objective in antifibrotic therapeutic strategies. At first only foetal cardiac fibroblasts were taken into consideration, then the study developed comparing normoglycaemic versus hyperglycaemic patients derived cells. The results of this research provide evidence for a novel class of TET2-selective non-nucleoside analogues capable of modulating DNA methylation. These compounds hold the potential to become valuable tools in the prevention and reduction of cardiac fibrosis.È consentito all'utente scaricare e condividere i documenti disponibili a testo pieno in UNITESI UNIPV nel rispetto della licenza Creative Commons del tipo CC BY NC ND.
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https://hdl.handle.net/20.500.14239/16510