Cystic fibrosis is an autosomal recessive disease that affects 1:3000 people in Australasia. It is caused by a mutation in the CFTR gene with over 2000 different mutations currently known. ΔF508 and W1282X are two examples. The misfolded or mis-trafficked CFTR protein causes dehydrated and sticky mucus in the lungs which inhibits mucociliary clearance. Viruses such as rhinovirus, which cause the common cold, can present as a more serious illness in people with cystic fibrosis.
Impact
Even with the advent of CFTR modulators, which are medications predominantly useful for people with the ΔF508 form of the gene, there is still no cure for cystic fibrosis. A cure would likely require gene therapy, particularly for patients who are ineligible for CFTR modulators, such as those with the W1282X form. People with cystic fibrosis are often much more susceptible to common respiratory pathogens, and it has been unclear whether this is directly due to the CFTR mutation, or if this is because of long-term stress and damage of the lungs. To explore this, we can use human induced pluripotent stem cells (hiPSCs) harbouring common CFTR mutations alongside an isogenic control. To model viral infection, these iPSCs are differentiated into airway basal cells and then plated at an Air-Liquid Interface (ALI) to mature into a pseudostratified airway epithelium, before being infected with respiratory viruses.
reNEW research
Our team has generated iPSC-derived basal cells harbouring two different mutations, ΔF508 and W1282X, as well as a non-CF counterpart. The basal cells can be plated at air-liquid interface (ALI) to differentiate into airway epithelial cells, such as goblet cells, club cells and ciliated cells. With this model we can explore how the mutations respond to rhinovirus (common cold) infections in comparison to the non-CF control. We are currently exploring whether CFTR modulators or gene therapy delivered with nanoparticles can restore infection responses in iPSC-derived ΔF508 and W1282X airway epithelium.
Image description
iPSC-derived basal cells have been plated at air-liquid interface (ALI). This culture has matured into a pseudostratified airway epithelium made up of goblet cells, club cells and ciliated cells. This iPSC cell line harbours a common CFTR mutation called W1282X, which forms a truncated and defective CFTR protein. The tight junctions between the cells are shown in green, with the nuclei of cells visible in blue. We can see club cells in magenta and the spindly ‘fingers’ of the cilia are in red. These hair-like cilia ‘beat’ to physically move mucus through the airways.
Credits
Katelyn Patatsos, Research Assistant, Werder Lab, reNEW Melbourne.
