Fibrosis is commonly associated with high morbidity and mortality, indeed, nearly 45% of all deaths in the developed world are attributed to chronic fibroproliferative diseases. Strong evidence indicates that chronic inflammation triggers fibrosis that, once established, may progress independently.
Serious complications of Crohn¿s disease (CD), a form of Inflammatory Bowel Disease (IBD), involves the formation of intestinal fibrotic stenosis followed by obstruction.
Currently, a comprehensive understanding of the pathogenesis of gut fibrosis in CD remains uncertain hampering the development of effective therapeutic strategies.
Patient-derived organoids represent a promising novel platform for the study of the development and progression of fibrosis (fibrostenosis) in CD patients.
In the present project, we aim: 1) to induce the inflammation-mediated fibrosis in human intestinal organoids (colonoids), whose generation has been recently set up in our laboratory, from intestinal stem cells originated from isolated intestinal crypts of the colon of CD patients, by exposing them to pro-inflammatory (TNFalpha) and pro-fibrotic (TGFbeta) agents; 2) to confirm in colonoids the role of the transcription factor ZNF281 as a promoter of gut fibrosis, previously shown in established intestinal cell lines; 3) to explore in colonoids the potential of non-pharmacological agents, in particular the beta 18-glycirrethic acid, the main metabolite of glycyrrhizin obtained from licorice root, and the extracts from Scutellaria baicalensis Georgi and Boswellia serrata, alone or in combination, as anti-fibrotic drugs.
This is a translational and innovative research project that will lead to remarkable advances in the scientific knowledge of CD fibrosis and management. The novelty of this study lies in the set-up of fibrotic colonoids as a novel tool to investigate the role of new protagonists of the intestinal fibrotic process and the identification of novel therapies for its treatment.
Fibrosis is commonly associated with high morbidity and mortality, indeed, nearly 45% of all deaths in the developed world are attributed to chronic fibroproliferative diseases. Strong evidence indicates that chronic inflammation triggers fibrosis that, once established, may progress independently.
The incidence of fibrosis-related complications in CD has not markedly changed over the last ten years, suggesting that the progression of intestinal fibrosis may be at least in part independent from the control of the underlying inflammatory disease.
Although mechanisms underlying CD have been recently deeply investigated, a comprehensive understanding of the pathogenesis of gut fibrosis remains uncertain hampering the development of effective therapeutic strategies.
Patient-derived organoids represent a promising novel platform for the study of the development and progression of fibrostenosis in CD patients. Really, organoids have the potential to bridge the gap between monolayer cell culture and whole-organism environments. Indeed, they are derived from human tissue and recapitulate the complex cellular organization seen in-vivo. They also are less-costly and avoid the issues of xenogeneity which may be associated with animal models. Really, patient-derived organoids may have translational applications in the future to understand mechanisms underlying human diseases as well as to find new strategies for its cure.
In this study, we will induce for the first time the inflammation-derived fibrosis in human intestinal organoids (colonoids) from stem cells originated from isolated intestinal crypts of the colon of CD patients, by a co-exposure to pro-inflammatory (TNF-alpha) and pro-fibrotic (TGF-beta) agents. The induction of fibrosis in human colonoids will allow to better and deeply investigate the onset of the fibrotic process in intestinal tissue by focusing on novel protagonists of fibrogenesis such as the transcription factor ZNF281, an epithelial to mesenchymal (EMT) inducer, that we have recently shown to play a main role in gut fibrosis in established epithelial cell lines. After the TGF-beta-mediated induction of fibrosis, ZNF281 will be silenced for the first time in colonoids and the expression levels of fibrotic markers, such as alpha-SMA and others, will be analyzed. This will allow to deeply explore the complicated network underlying ZNF281-mediated gut fibrosis and will open new perspective for its management.
Then, we will also provide novel information about the potential of some compounds of natural origin to function as anti-fibrotic agents in agreement with preliminary data we obtained in established cell lines. Indeed, we recently showed that the beta 18-glycirrethic acid (GA) is a strong anti-inflammatory compound. As well, several literature data suggest that it exerts anti-fibrotic effects in different human organs, including the liver. Moreover, preliminary data obtained in our laboratory demonstrate that the extracts from Scutellaria baicalensis Georgi (SBG) and Boswellia serrata (BS), alone or in combination, may reduce fibrosis in intestinal cell lines (HIEC). The identification of novel agents able to improve gut fibrosis will help clinicians, who today complain of the lack of therapies aimed at resolving fibrosis in patients with chronic inflammatory diseases.
In conclusion, this translational research project promises to be very innovative, since it will lead to remarkable advances in the scientific knowledge of CD fibrosis and management. The novelty of this study lies in the set up of fibrotic colonoids as a novel tool to investigate the onset of the gut fibrotic process as well as the identification of novel therapies for its treatment.