A new platform for high-throughput therapy testing on iPSC-derived, immature airway from Cystic Fibrosis Patients

Abstract

Induced pluripotent, stem cell (iPSC)-derived models of airway tissue have successfully modeled the primary defect in regulated chloride conductance caused by the major Cystic Fibrosis causing mutation, F508del. However, it remains unclear if iPSC-derived airway cultures can be used in high-throughput therapy development for F508del and rarer mutations. There is an urgent need for airway tissue models that reflect the variability of patient-specific responses and are scalable for therapy development. In the current work, we describe a robust, high-throughput fluorescence assay of mutant CFTR function in iPSCs differentiated to immature airway epithelium. This assay measures reproducible functional responses to modulators targeting either the major CF mutant F508del or the nonsense mutant: W1282X-CFTR. We show that the ranking of patient-specific responses to interventions in this stem-cell based model recapitulates the ranking observed in primary nasal epithelial cultures obtained from the same individuals. In summary, these proof-of-concept studies show that this novel platform has the potential to support therapy development and precision medicine for Cystic Fibrosis patients.

Modeling lung cell development using human pluripotent stem cells

“…Both hPSC lines demonstrated enhanced CFTR function averaging around 32% function in our new protocol compared to our old (~10% function) (Fig. 3F) and thus provide a powerful source of cells for rapid high throughput functional screens of novel CFTR modulators 37 .…”

“…Importantly, hPSC-derived AS provides a user-friendly opportunity that is time and cost-effective for researchers with no experience or capacity to work with hPSC. Their proliferative and relatively undifferentiated nature enables rapid expansion of AS for regenerative studies 13 and developing high throughput assays for disease models and/or drug screens 37 . Upon differentiation, the airway epithelial organoids recapitulate proximal airway phenotype with basal cells on the apical side with mature cell types (secretory, ciliatory) and CFTR localized to the luminal side.…”

Perspectives on the translation of in-vitro studies to precision medicine in Cystic Fibrosis

“…iPSC differentiation protocols which result in more mature tissue need to be developed. However existing iPSC derived immature lung models are still able to predict patient specific response to drug modulators [62]. Other differentiation protocols focusing on the differentiation of iPSCs to basal cells have been developed enabling the in-vitro modeling of airway diseases [63].…”

Modeling lung cell development using human pluripotent stem cells

“…The ACC assay was used to assess CFTR mediated changes in membrane depolarization using methods as previously described 37 . iPSC-derived airway cultures were replated on collagen IV (Sigma Aldrich, cat# C7521)-coated 96 well plates (Corning, cat# 3603) at AFVE stage Day 3.…”