Coupling single cell insights in airway epithelial biology to targeted gene editing delivery to treat the cystic fibrosis gene defect

Cystic fibrosis is a progressive, hereditary, and severe disease that is fatal and estimated to affect the lives of more than 160,000 people worldwide. The disease is caused by various loss-of-function mutations in both alleles of the CFTR gene. These mutations cause the CFTR ion channel to be defective, leading to a disrupted water transport in various organs. In the lungs, this water-salt imbalance results in chronic inflammation and infections. While treatments exist to help 90% of patients, strategies are being developed for the more diverse CFTR mutations, and as an alternative to current treatments, which require intake twice a day and are not a cure. Various CRISPR-Cas based techniques are on the rise to remedy various mutations. Nevertheless, the bottleneck of these therapies lies in delivering the components to the correct target cell types, which could consist of differentiated cell types with a high CFTR expression such as secretory cell types (club & goblet cells) and ionocytes, as well as of basal cells, which are known as progenitor cells. Besides keeping the original biological function of the target cell types, a lack of, or a minimum amount of adverse effects is preferable. Prolonged exposure of CRISPR-Cas based components to tissues can lead to unwanted modifications, indicating the importance to opt for a transient delivery vehicle able to perform targeted gene correction. This research focuses on: (1) identification of suitable target proteins on the plasma membrane of human airways, (2) the development or discovery of suitable proteins for delivery vehicle pseudotyping (binding partners of the target proteins) and (3) the validation of the efficiency and specificity of these pseudotyped delivery modalities in primary human tissue. Additionally, the research assesses whether the biological function is conserved within the target cell populations and evaluates any potential adverse effects, which would be caused by gene editing.