Investigation of CFTR Function in Human Nasal Epithelial Cells Informs Personalized Medicine

Am J Respir Cell Mol Biol. 2024 Nov;71(5):577-588. doi: 10.1165/rcmb.2023-0398OC.

Abstract

We broaden the clinical versatility of human nasal epithelial (HNE) cells. HNEs were isolated from 10 participants harboring cystic fibrosis transmembrane conductance regulator (CFTR) variants: 9 with rare variants (Q359R [n = 2], G480S, R334W [n = 5], and R560T) and 1 harboring R117H;7T;TG10/5T;TG12. Cultures were differentiated at the air-liquid interface. CFTR function was measured in Ussing chambers at three conditions: baseline, ivacaftor, and elexacaftor + tezacaftor + ivacaftor (ETI). Four participants initiated modulators. Q359R HNEs had 5.4% (% wild-type) baseline CFTR function and 25.5% with ivacaftor. With therapy, sweat [Cl-] decreased and symptoms resolved. G480S HNEs had 4.1% baseline and 32.1% CFTR function with ETI. Clinically, forced expiratory volume in 1 second increased and sweat [Cl-] decreased (119 to 46 mmol/L) with ETI. In vitro cultures derived from 5 participants harboring R334W showed a moderate increase in CFTR function with exposure to modulators. For one of these participants, ETI was begun in vivo; symptoms and forced expiratory volume in 1 second improved. The c.1679G>C (R560T) HNEs had less than 4% baseline CFTR function and no modulator response. RNA analysis confirmed that c.1679G>C completely missplices. A symptomatic patient harboring R117H;7T;TG10/5T;TG12 exhibited reduced CFTR function (17.5%) in HNEs, facilitating a diagnosis of mild CF. HNEs responded to modulators (ivacaftor: 32.8%, ETI: 55.5%), and, since beginning therapy, lung function improved. We reaffirm HNE use for guiding therapeutic approaches, inform predictions on modulator response (e.g., R334W), and closely assess variants that affect splicing (e.g., c.1679G>C). Notably, functional studies in HNEs harboring R117H;7T;TG10/5T;TG12 facilitated a diagnosis of mild CF, suggesting the use for HNE functional studies as a clinical diagnostic test.

Keywords: 5T; air–liquid interface; cystic fibrosis; modulators; nasal epithelial.

MeSH terms

  • Adult
  • Aminophenols* / pharmacology
  • Benzodioxoles / pharmacology
  • Cells, Cultured
  • Chloride Channel Agonists / pharmacology
  • Cystic Fibrosis Transmembrane Conductance Regulator* / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator* / metabolism
  • Cystic Fibrosis* / drug therapy
  • Cystic Fibrosis* / genetics
  • Cystic Fibrosis* / metabolism
  • Drug Combinations
  • Epithelial Cells* / drug effects
  • Epithelial Cells* / metabolism
  • Female
  • Humans
  • Indoles / pharmacology
  • Male
  • Middle Aged
  • Mutation
  • Nasal Mucosa* / drug effects
  • Nasal Mucosa* / metabolism
  • Precision Medicine* / methods
  • Pyrazoles / pharmacology
  • Pyridines / pharmacology
  • Pyrroles / pharmacology
  • Quinolones* / pharmacology
  • Sweat / metabolism
  • Young Adult

Substances

  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Quinolones
  • Aminophenols
  • ivacaftor
  • CFTR protein, human
  • Indoles
  • Benzodioxoles
  • Pyrroles
  • Chloride Channel Agonists
  • Pyrazoles
  • Pyridines
  • Drug Combinations
  • tezacaftor