Identification of novel bacterial virulence factors and inflammation determinants associated with persistent lung infections in Cystic Fibrosis
2020 – 2025
Overview
Chronic lung infections caused by Pseudomonas aeruginosa are a leading cause of mortality and reduced quality of life in cystic fibrosis (CF) patients. Despite advances in medical treatment, eradicating this pathogen remains a significant challenge. This innovative research project, generously funded by Fondazione Cariplo, focuses on uncovering the molecular and physiological mechanisms that enable P. aeruginosa to persist in the hostile environment of CF airways, drive inflammation, and evade host defenses. By bridging cutting-edge molecular biology, advanced infection models, and translational approaches, the project aims to redefine treatment strategies for CF-associated infections.
Aims
This research strives to:
- Decipher Pathogenic Mechanisms: Identify and characterize the genetic and metabolic pathways that support P. aeruginosa persistence in the unique environment of CF airways.
- Understand Host-Pathogen Interactions: Explore how P. aeruginosa interacts with the host’s immune system to sustain chronic infections and provoke an exaggerated inflammatory response.
- Develop New Therapeutic Strategies: Use insights from the research to identify novel molecular targets for therapies aimed at reducing bacterial persistence and inflammation.
- Advance Experimental Models: Establish robust infection models using human-derived airway organoids that closely mimic CF conditions.
Why This Project Matters?
Despite decades of research, P. aeruginosa infections in CF remain resistant to eradication, primarily due to gaps in understanding the pathogen’s physiology in vivo. This project bridges these gaps by:
- Targeting the actual processes occurring in CF airways, rather than relying solely on in vitro models.
- Shedding light on the paradoxical inflammatory responses triggered by less virulent bacterial forms.
- Offering translational solutions that can be quickly adapted to clinical practice, including drug development and diagnostic tools.
Methods
To achieve its goals, the project employs a comprehensive and interdisciplinary approach:
- Gene Expression Profiling:
- Using transcriptomics, the project investigates P. aeruginosa gene expression directly in patient-derived sputum samples to capture real-time bacterial responses in vivo.
- Identification of differentially expressed genes (DEGs) that regulate persistence and immune evasion.
- Mutant Strain Library:
- A library of P. aeruginosa knockout (KO) and overexpression (OEXP) mutants is created to test their roles in biofilm formation, stress resistance, and virulence.
- RNA sequencing is performed on these mutants to uncover system-wide physiological changes and regulatory networks.
- Human Airway Organoids:
- Patient-derived human airway organoids (HAOs) are used to replicate CF airway conditions. These three-dimensional models provide insights into colonization, tissue damage, and immune response.
- Organoids are infected with mutant strains to evaluate bacterial persistence, inflammatory response, and neutrophil recruitment.
- Collaborative Framework:
- The project benefits from the expertise of two research units: the University of Milan, Italy, and Rigshospitalet, Copenhagen, Denmark. This collaboration ensures a combination of molecular techniques and advanced infection models.
- Advanced Imaging Techniques:
- Confocal microscopy and time-lapse imaging are utilized to visualize bacterial colonization and immune cell recruitment in organoid models, offering dynamic insights into host-pathogen interactions.
- Data Analysis and Validation:
- A robust statistical framework and machine learning algorithms are applied to analyze complex datasets, ensuring reproducibility and reliability of findings.
Expected Outcomes
Scientific Impact:
- PaEnhanced Understanding: Gain a deeper knowledge of the metabolic and genetic adaptations of P. aeruginosa in CF airways.
- Target Identification: Identify key molecular pathways and genes that can serve as potential targets for therapeutic intervention.
- Model Development: Establish HAOs as a reliable and translatable model for studying CF-related infections.
Societal Relevance:
- Better Treatments: Provide a foundation for developing precision medicine approaches that target P. aeruginosa persistence mechanisms, improving outcomes for CF patients.
- Reduced Healthcare Burden: Alleviate the impact of chronic infections and associated inflammation on healthcare systems and patient quality of life.
- Public Awareness and Engagement: Through active dissemination efforts, educate patients, clinicians, and the broader public about advancements in CF treatment.
Partners
Fundings
