Carbonic Anhydrase Mediated Regulation of Microtubule Dynamics and Infection in Cystic Fibrosis

Project Summary Cystic fibrosis (CF) is a genetic disease causing premature death, primarily due to respiratory failure from increased inflammation and chronic infections. While therapies targeting the dysfunctional protein channel have improved lung function and mucus clearance, infection remains a significant problem. This proposal investigates how impaired microtubule (MT) dynamics and intracellular transport, exacerbated by reduced carbonic anhydrase (CA) expression, contribute to increased bacterial adhesion in CF. The central hypothesis is that reduced CA expression disrupts MT dynamics, leading to impaired intracellular transport and accumulation of pathogen-binding targets on cell membranes. The approach will use advanced live cell imaging, biochemical assays, proteomics, and microbial adherence studies to test hypotheses across three specific aims: 1) Determine the role of CA2 and CA12 in intracellular transport and membrane organization; 2) Identify the binding target and mechanism for increased bacterial adhesion in CA-deficient airway epithelia; and 3) Assess the roles of MTs and CA in the ability to eradicate Pseudomonas aeruginosa among people with CF, determining in vitro response to MT- and CA-targeted agents. This research will provide new insights into the cellular mechanisms of infection in CF and could identify novel therapeutic targets to improve infection control. The significance of this study lies in its comprehensive approach to understanding and addressing infection in CF, which can be applied to other diseases involving defective intracellular transport and cytoskeletal dynamics. This project will also serve as an excellent vehicle for the applicant’s development into an independent investigator. Investigations will be performed in an environment with an established history of successful mentorship of junior faculty to independence. The applicant’s long-term career goal is to become a leading, independent investigator with a translational lab studying cellular mechanisms driving infection and inflammation in the lungs, with a specific focus on host-microbial interactions that modify the landscape of microbiota within the airway. With the support of this award, the applicant will 1) build technical skills in advanced imaging techniques, 3D model systems, and bioinformatics, and 2) develop expertise in interpreting data and translating findings to clinical therapeutics. Future independent studies will focus on the relevant pathways and targets affected by CA and MT disruption to identify possible therapeutic interventions that can be tested in pre-clinical, and ultimately clinical, models.