In vitro models to study the role of N6-methyladenosine in human pulmonary inflammation

In vitro models to study the role of N6-methyladenosine in human pulmonary inflammation

Project code: Bulgarian National Science Fund КП-06-Рила/1
Call: Franco-Bulgarian Hubert Curien Partnership (PHC) Program RILA-2025
Duration: 01.01.2026 - 31.12.2027
Budget: 25, 564.60 euro
Collaboration: Dr. Milena LESEVA, Laboratory of Experimental Immunotherapy – Stephan Angeloff Institute of Microbiology (SAIM); Sofia, Bulgaria and Dr. Alexandre GRASSART, Mechano-Biology of Host-Microbe Interactions – Center for Infection and Immunity of Lille (CIIL); Lille, France
Pulmonary inflammation is integral to the pathogenesis of many chronic diseases of the lower airways, including interstitial lung disease (ILD). Interstitial lung diseases are a heterogeneous group of >200 different clinical disorders, which are associated with physical disability and a severely decreased quality of life due to progressive lung tissue damage. Among ILDs, fibrotic interstitial lung disease (fILD) is the most severe, with a 5-year survival rate of <50%. Currently available models used to study fILDs are mostly restricted to experimental in vivo models using small laboratory animals. This pilot project will investigate the role of internal RNA modification, specifically N6-methyladenosine, during the pre-fibrotic inflammatory stage of fibrotic interstitial lung disease. Specifically, we will study whether METTL3/14-mediated RNA methylation: 1) exacerbates the innate immune response to inflammatory injury in the alveolar compartment; and 2) accelerates the emergence of a senescence-associated secretory phenotype (SASP). In order to achieve this, we will establish an advanced in vitro model of human alveolar inflammation using the alveolus-on-chip and a model of bronchial epithelium inflammation using air-liquid-interface differentiated normal human bronchial epithelial cells.
Expected results: In a microfluidic alveolus-on-chip system, we expect that direct treatment with the pro-inflammatory cytokine IL1β or indirect induction of inflammation using cells from patients with pulmonary fibrosis will increase the expression of mediators of fibrosis in the alveolar epithelium. In an in vitro differentiated model of human bronchiolar epithelium, we expect an increased expression of pro-inflammatory cytokines after induction with IL1β. We expect to be able to identify genes for cytokines/chemokines or epigenetic modifiers the expression level of which changes after induction of inflammatory injury, and which can serve as its molecular markers. We expect an exacerbation of inflammatory injury following activation of METTL3 and that repeated inflammatory injury in the models of alveolar and bronchiolar epithelium increases the expression of SASP genes. It is likely that activation and inhibition of METTL3 affects the temporal dynamics of this process.