Human Amnion Epithelial Cells Induced to Express Functional Cystic Fibrosis Transmembrane Conductance Regulator

Sean V Murphy,Daryl L Howard,Courtney Mcdonald,Philip Heraud,Rebecca Lim,Marian Cholewa,Anthony Atala,Euan M Wallace,Martin D De Jonge,Mark Le Gros,Graham Jenkin,David Paterson

doi:10.1371/journal.pone.0046533

Abstract

Cystic fibrosis, an autosomal recessive disorder caused by a mutation in a gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), remains a leading cause of childhood respiratory morbidity and mortality. The respiratory consequences of cystic fibrosis include the generation of thick, tenacious mucus that impairs lung clearance, predisposing the individual to repeated and persistent infections, progressive lung damage and shortened lifespan. Currently there is no cure for cystic fibrosis. With this in mind, we investigated the ability of human amnion epithelial cells (hAECs) to express functional CFTR. We found that hAECs formed 3-dimensional structures and expressed the CFTR gene and protein after culture in Small Airway Growth Medium (SAGM). We also observed a polarized CFTR distribution on the membrane of hAECs cultured in SAGM, similar to that observed in polarized airway cells in vivo. Further, hAECs induced to express CFTR possessed functional iodide/chloride (I−/Cl−) ion channels that were inhibited by the CFTR-inhibitor CFTR-172, indicating the presence of functional CFTR ion channels. These data suggest that hAECs may be a promising source for the development of a cellular therapy for cystic fibrosis.

Highlights

The need for a cure of cystic fibrosis remains as urgent today as ever
CFTR expression and function is essential for normal respiratory airway function and is defective in individuals with cystic fibrosis
With a view to developing a cell-based therapy for cystic fibrosis we have shown that human amnion epithelial cells can be induced to express functional cystic fibrosis transmembrane conductance regulator (CFTR) in vitro by culture in media designed for lung cell differentiation

Summary

Introduction

Since the discovery of the CFTR gene and the mutations that cause cystic fibrosis, attempts at gene correction by CFTR gene transfer to the airways, using both viral and non-viral gene-transfer mechanisms, have been described [1,2,3]. This approach to treatment, at least to date, has been unsuccessful. Cells isolated from the human placenta and fetal membranes have considerable advantages over other sources of stem cells such as embryonic or bone marrow-derived cells. We report the ability to induce hAECs to express functional CFTR in vitro, providing a valuable tool for a potential cellular therapy for cystic fibrosis

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Conclusion