Primary Human Nasal Epithelial Cells: Biobanking in the Context of Precision Medicine

Mairead Kelly; Elise Dreano; Aurelie Hatton; Agathe Lepissier; Anita Golec; Isabelle Sermet-Gaudelus; Iwona Pranke

doi:10.3791/63409

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Method Article

Primary Human Nasal Epithelial Cells: Biobanking in the Context of Precision Medicine

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DOI:

10.3791/63409

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April 22nd, 2022

Mairead Kelly¹^,² , Elise Dreano¹^,² , Aurelie Hatton¹^,² , Agathe Lepissier¹^,² , Anita Golec¹^,² , Isabelle Sermet-Gaudelus¹^,²^,³ , Iwona Pranke¹^,²^,³

¹Institut Necker Enfants Malades, ²Université de Paris, ³Centre de Référence Maladies Rares Mucoviscidose et Maladies apparentées, Assistance Publique Hôpitaux de Paris

Summary

Here we describe the isolation, amplification, and differentiation of primary human nasal epithelial (HNE) cells at the air-liquid interface and a biobanking protocol allowing to successfully freeze and then thaw amplified HNE. The protocol analyzes electrophysiological properties of differentiated HNE cells and CFTR-related chloride secretion correction upon different modulator treatments.

Abstract

Human nasal epithelial (HNE) cells are easy to collect by simple, non-invasive nasal brushing. Patient-derived primary HNE cells can be amplified and differentiated into a pseudo-stratified epithelium in air-liquid interface conditions to quantify cyclic AMP-mediated Chloride (Cl^-) transport as an index of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) function. If critical steps such as quality of nasal brushing and cell density upon cryopreservation are performed efficiently, HNE cells can be successfully biobanked. Moreover, short-circuit current studies demonstrate that freeze-thawing does not significantly modify HNE cells' electrophysiological properties and response to CFTR modulators. In the culture conditions used in this study, when less than 2 x 10⁶ cells are frozen per cryovial, the failure rate is very high. We recommend freezing at least 3 x 10⁶ cells per cryovial. We show that dual therapies combining a CFTR corrector with a CFTR potentiator have a comparable correction efficacy for CFTR activity in F508del-homozygous HNE cells. Triple therapy VX-445 + VX-661 + VX-770 significantly increased correction of CFTR activity compared to dual therapy VX-809 + VX-770. The measure of CFTR activity in HNE cells is a promising pre-clinical biomarker useful to guide CFTR modulator therapy.

Introduction

Cystic Fibrosis (CF) is an autosomal recessive disorder resulting from mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene leading to the absence or dysfunction of the CFTR protein, an anion channel located at the apical surface of epithelia¹^,². Recent advances in CFTR therapy have improved the prognosis of the disease, and the last approved drugs combining CFTR correctors and CFTR potentiators led to major improvements in lung function and quality of life for CF patients carrying the most frequent mutation p.Phe508del mutation (F508del)³^,