Epithelial disruption: a new paradigm enabling human airway stem cell transplantation

Nigel Farrow,Nathan Rout-Pitt,Yuben Moodley,David Parsons,Ivan Bertoncello,Martin Donnelley,Patricia Cmielewski

doi:10.1186/s13287-018-0911-4

Abstract

BackgroundAirway disease is a primary cause of morbidity and early mortality for patients with cystic fibrosis (CF). Cell transplantation therapy has proven successful for treating immune disorders and may have the potential to correct the airway disease phenotype associated with CF. Since in vivo cell delivery into unconditioned mouse airways leads to inefficient engraftment, we hypothesised that disrupting the epithelial cell layer using the agent polidocanol (PDOC) would facilitate effective transplantation of cultured stem cells in mouse nasal airways.MethodsIn this study, 4 μL of 2% PDOC in phosphate-buffered saline was administered to the nasal airway of mice to disrupt the epithelium. At 2 or 24 h after PDOC treatment, two types of reporter gene-expressing cells were transplanted into the animals: luciferase-transduced human airway basal cells (hABC-Luc) or luciferase-transduced human amnion epithelial cells (hAEC-Luc). Bioluminescence imaging was used to assess the presence of transplanted luciferase-expressing cells over time. Data were evaluated by using two-way analysis of variance with Sidak’s multiple comparison.ResultsSuccessful transplantation was observed when hABCs were delivered 2 h after PDOC but was absent when transplantation was performed 24 h after PDOC, suggesting that a greater competitive advantage for the donor cells is present at the earlier time point. The lack of transplantation of hAECs 24 h after PDOC supports the importance of choosing the correct timing and cell type to facilitate transplantation.ConclusionsThese studies into factors that may enable successful airway transplantation of human stem cells showed that extended functioning cell presence is feasible and further supports the development of methods that alter normal epithelial layer integrity. With improvements in efficacy, manipulating the airway epithelium to make it permissive towards cell transplantation may provide another option for safe and effective correction of CF transmembrane conductance regulator function in CF airways.

Highlights

Airway disease is a primary cause of morbidity and early mortality for patients with cystic fibrosis (CF)
Vector transduction of Human airway basal cell (hABC) and Human amnion epithelial cell (hAEC) Prior to the reporter gene transduction procedures, hABC cultures were confirmed as 95% pure basal cells via Cytokeratin 5 (Krt5) staining. hABCs and hAECs were expanded in culture and transduced with an LV vector carrying the
LacZ or Luc reporter gene at an Multiplicity of infection (MOI) of 10 (Fig. 1). hABC-LacZ were cultured for 4 weeks, and X-gal staining showed that the cells remained healthy and viable and that LacZ transgene expression was sustained for this period (Fig. 1a)

Summary

Introduction

Airway disease is a primary cause of morbidity and early mortality for patients with cystic fibrosis (CF). Cystic fibrosis (CF) is an autosomal recessive genetic disorder that affects multiple organs, but the main cause of morbidity and early mortality is lung disease [1]. The cystic fibrosis transmembrane conductance regulator (CFTR) protein is a chloride channel on the surface of epithelial cells. The cilia that are a part of the epithelial architecture and are required to clear pathogens from the airway are immobilised in a milieu of highly viscous mucous [3]. This provides an airway environment amenable to colonisation by bacterial and viral pathogens. The lung disease that is associated with the CF airway phenotype is responsible for the roughly 40-year life expectancy of the patient with CF

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