Detection of CFTR function and modulation in primary human nasal cell spheroids

J Cyst Fibros. 2018 Jan;17(1):26-33. doi: 10.1016/j.jcf.2017.06.010. Epub 2017 Jul 13.

Abstract

Background: Expansion of CFTR modulators to patients with rare/undescribed mutations will be facilitated by patient-derived models quantifying CFTR function and restoration. We aimed to generate a personalized model system of CFTR function and modulation using non-surgically obtained nasal epithelial cells (NECs).

Methods: NECs obtained by curettage from healthy volunteers and CF patients were expanded and grown in 3-dimensional culture as spheroids, characterized, and stimulated with cAMP-inducing agents to activate CFTR. Spheroid swelling was quantified as a proxy for CFTR function.

Results: NEC spheroids recapitulated characteristics of pseudostratified respiratory epithelia. When stimulated with forskolin/IBMX, spheroids swelled in the presence of functional CFTR, and shrank in its absence. Spheroid swelling quantified mutant CFTR restoration in F508del homozygous cells using clinically available CFTR modulators.

Conclusions: NEC spheroids hold promise for understanding rare CFTR mutations and personalized modulator testing to drive evaluation for CF patients with common, rare or undescribed mutations. Portions of this data have previously been presented in abstract form at the 2016 meetings of the American Thoracic Society and the 2016 North American Cystic Fibrosis Conference.

Keywords: CFTR; Modulator; Organoid; Personalized model system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Cell Culture Techniques / methods*
  • Child
  • Child, Preschool
  • Chloride Channel Agonists / pharmacology
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism*
  • Cystic Fibrosis* / genetics
  • Cystic Fibrosis* / metabolism
  • Cystic Fibrosis* / pathology
  • Epithelial Sodium Channels / metabolism*
  • Female
  • Humans
  • Infant
  • Male
  • Mutation
  • Nasal Mucosa* / metabolism
  • Nasal Mucosa* / pathology

Substances

  • Chloride Channel Agonists
  • Epithelial Sodium Channels
  • Cystic Fibrosis Transmembrane Conductance Regulator