Cholinergic Signaling Modulates Intestinal Pathophysiology in a Drosophila Model of Cystic Fibrosis

Cystic Fibrosis (CF) is a genetic disease caused by mutations in the CFTR gene, which affects cells in organs like the lungs, pancreas, and gut. Treatments aim to fix the CFTR protein, but many gut issues remain untreated. In this study, we use fruit flies (Drosophila) as a model to study the effects of losing CFTR specifically in the gut. We show that knocking down CFTR in gut cells leads to problems seen in CF, such as poor gut movement, trouble absorbing nutrients, and low energy levels. Using RNA sequencing, we find that CF guts have higher levels of the gene acetylcholine esterase (Ace), which lowers cholinergic (nerve signal) activity. Experiments confirm that CF guts are less sensitive to cholinergic signals, but reducing Ace brings back normal gut function. We also find a transcription factor called Forkhead (Fkh), similar to human FOXA1/2, that increases Ace in CF. Our study shows that the fruit fly gut is a useful model for studying CF in the gut, and suggests that boosting cholinergic signaling could help treat gut problems in CF. Overall design: Fruit flies (Drosophila) was used to perform enterocyte-specific knockdown of CFTR gene to model CF gut pathology. Gut movement, nutrient absorption, and energy levels were assessed. Single-nuclei RNA sequencing (snRNA-seq) was performed to identify gene expression changes, revealing increased expression of acetylcholine esterase (Ace). Functional assays tested cholinergic signaling and showed that Ace knockdown could restore gut function. Finally, the role of the transcription factor Forkhead (Fkh) in regulating Ace was investigated.