Intestinal current measurement detects age-dependent differences in CFTR function in rectal epithelium

Simon Y. Graeber ^1,2,3* Olaf Sommerburg ^4,5 Yin Yu ^4,5 Stephanie Hirtz ^4,5 Heike Scheuermann ^4,5 Jasmin Berger ^1,2,3 Julia Duerr ^1,2,3 Marcus A Mall ^1,2,3

• ¹ Charité Universitätsmedizin Berlin, Department of Pediatric Respiratory Medicine, Immunology and Critical Care Medicine, Berlin, Baden-Württemberg, Germany
• ² German Center for Lung Research (DZL), associated partner site Berlin, Berlin, Baden-Württemberg, Germany
• ³ German Center for Child and Adolescent Health (DZKJ), partner site Berlin, Berlin, Baden-Württemberg, Germany
• ⁴ Division of Pediatric Pulmonology, Allergy and Cystic Fibrosis Center, Department of Pediatrics, University Hospital Heidelberg, Heidelberg, Germany
• ⁵ Translational Lung Research Center, Heidelberg University Hospital, Heidelberg, Baden-Württemberg, Germany

Objective: Intestinal current measurement (ICM) provides a sensitive bioassay for assessment of CFTR function in rectal biopsies ex vivo. Furthermore, ICM was shown to be sensitive to detect pharmacological rescue of CFTR function by CFTR modulators in people with CF. Results from clinical trials of CFTR modulators across age groups indicate that CFTR function in the sweat duct may be age-dependent. However, little is known about age dependency of CFTR function in the intestinal epithelium.Methods: We investigated CFTR-mediated chloride secretion in rectal biopsies from 258 people without CF and 72 people with pancreatic-insufficient CF from 1 month to 68 years of age. Change in transepithelial short-circuit current in response to cAMP-mediated and cholinergic stimulation was assessed using perfused micro-Ussing chambers. Furthermore, quantitative real-time PCR of CFTR and morphometric analysis of epithelial cells lining the crypts and surface of the rectal mucosa were performed.Results: We found that CFTR-mediated chloride secretion across rectal tissues, as determined from cAMP-mediated as well as cholinergic chloride-secretory responses was highest during infancy and early childhood and declined with age in people without CF. In people with CF, potassium-secretory responses induced by cholinergic stimulation were also reduced with increasing age. Transcript analyses showed that CFTR mRNA expression was slightly increased with increasing age in people without CF. Morphometric analyses demonstrated that CFTR expressing colonocytes at the crypt base were decreased with age. A secondary analysis of the ICM data of our previous studies on the effects of lumacaftor/ivacaftor on CFTR function in F508del -homozygous people with CF aged 12 years and older and 2 to 11 year old children showed correlations of the change in cAMP-mediated and cholinergic chloride secretory response with the age of people with CF.Conclusion: These results demonstrate that CFTR function in the rectal epithelium is reduced with increasing age and indicate that this change is likely due to a decline in the number of secretory colonocytes at the crypt base. These findings suggest that differences in CFTR expressing cells may explain increased functional responses to CFTR modulator therapies in children compared to adult people with CF.