INVESTING THE ROLES OF E26 TRANSFORMATION-SPECIFIC HOMOLOGOUS FACTOR IN CYSTIC FIBROSIS LUNG DISEASE
type: CreativeWork
description: CF (Cystic Fibrosis) is the most prevalent genetic disease among Caucasians. Here, a sodiumbicarbonate channel, the Cystic Fibrosis Transmembrane conductance Regulator (CFTR), is impaired, dehydrating all epithelia. The principal cause of morbimortality of CF is CF lung disease (CFLD) where CF airways have impaired mucociliary clearance and undergo constant remodelling and chronic inflammation, ultimately leading to irreversible loss of lung function. Interestingly, within CF patients carrying the same CFTR mutation resides a high variability in terms of CFLD severity and response to treatments. These variabilities are thought to be due to differentially expressed non-CFTR genes. To locate these genes, Genome-Wide Association Studies (GWAS) collected and analysed data from large patient cohorts, identifying four loci (5p15, 3q29, 11p12-13, Xq22-23) associated with more severe clinical manifestations of CFLD. These loci could sit close by genes potentially impacting CFLD penetrance, and patient responsiveness to therapeutics. We aim to understand how genes outlined by GWAS influence CFLD. To this end, we first optimized a model system which relies on human induced pluripotent stem cells (hiPSC)-derived Airways Epithelial Cells (AECs). We first explored two differentiation protocols that generate lung AECs as organoids and Air-Liquid-Interface (ALI) cultures and outlined their respective advantages and limitations. Then, using the most robust differentiation protocol, we explored the expression of GWAS-outlined genes and observed that E26 Transformation-Specific Homologous Factor (EHF) was expressed at comparable levels to primary Human Bronchial Epithelial Cells. Thus, we decided to study EHF in our AECs in-vitro model. First, we knocked out EHF using CRISPR-Cas9 from two non-CF hiPSC lines and selected, within the pool of edited cells, homozygous and heterozygous knock-out (KO) clones alongside wild-type clones, then used as internal controls. The KO was confirmed b…
abstract: CF (Cystic Fibrosis) is the most prevalent genetic disease among Caucasians. Here, a sodiumbicarbonate channel, the Cystic Fibrosis Transmembrane conductance Regulator (CFTR), is impaired, dehydrating all epithelia. The principal cause of morbimortality of CF is CF lung disease (CFLD) where CF airways have impaired mucociliary clearance and undergo constant remodelling and chronic inflammation, ultimately leading to irreversible loss of lung function. Interestingly, within CF patients carrying the same CFTR mutation resides a high variability in terms of CFLD severity and response to treatments. These variabilities are thought to be due to differentially expressed non-CFTR genes. To locate these genes, Genome-Wide Association Studies (GWAS) collected and analysed data from large patient cohorts, identifying four loci (5p15, 3q29, 11p12-13, Xq22-23) associated with more severe clinical manifestations of CFLD. These loci could sit close by genes potentially impacting CFLD penetrance, and patient responsiveness to therapeutics. We aim to understand how genes outlined by GWAS influence CFLD. To this end, we first optimized a model system which relies on human induced pluripotent stem cells (hiPSC)-derived Airways Epithelial Cells (AECs). We first explored two differentiation protocols that generate lung AECs as organoids and Air-Liquid-Interface (ALI) cultures and outlined their respective advantages and limitations. Then, using the most robust differentiation protocol, we explored the expression of GWAS-outlined genes and observed that E26 Transformation-Specific Homologous Factor (EHF) was expressed at comparable levels to primary Human Bronchial Epithelial Cells. Thus, we decided to study EHF in our AECs in-vitro model. First, we knocked out EHF using CRISPR-Cas9 from two non-CF hiPSC lines and selected, within the pool of edited cells, homozygous and heterozygous knock-out (KO) clones alongside wild-type clones, then used as internal controls. The KO was confirmed b…
author: Pinte, Laetitia
language: eng
keywords: Biology, Cystic Fibrosis, Genetic modifiers, Induced pluripotent stem cells, Lung disease, Stem Cells
license: https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/f8b84368-46fd-41cc-90a9-396e0feb4e3a/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2023-04-01
date modified: 2026-07-22T22:24:07Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Cambridge university

PropertyValue
property ID: DOI
value: 10.17863/CAM.99854