Comparison of ex vivo and in vitro intestinal cystic fibrosis models to measure CFTR-dependent ion channel activity

Ommen, Domenique D. Zomer-van; Poel, Eyleen de; Kruisselbrink, Evelien; Oppelaar, Hugo; Vonk, Annelotte M.; Janssens, Hettie M.; Ent, Cornelis K. van der; Hagemeijer, Marne C.; Beekman, Jeffrey M.

doi:10.1016/j.jcf.2018.02.007

Cited by 39 publications

(35 citation statements)

References 34 publications

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“…Patient-specific mechanisms that control F508del protein maturation and apical trafficking, which are not maintained in the organoid culture, may also contribute [46], as may the impact of cAMP/Ca 2+ -stimulated signalling pathways on non-CFTR chloride channels [48]. As the organoid culture conditions enrich for the crypt-based secretory stem cell epithelial compartment of the intestinal epithelium, and not the whole intestinal epithelium, the ICM-based observations may result from currents evoked by intestinal cells that are reduced in the organoid cultures, such as goblet cells [49]. Clearly, additional investigations beyond the scope of the present study are needed to understand these interesting findings.…”

Section: Discussionmentioning

confidence: 99%

Stratifying infants with cystic fibrosis for disease severity using intestinal organoid swelling as a biomarker of CFTR function

et al. 2018

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Forskolin-induced swelling (FIS) of intestinal organoids from individuals with cystic fibrosis (CF) measures function of the cystic fibrosis transmembrane conductance regulator (CFTR), the protein mutated in CF.We investigated whether FIS corresponds with clinical outcome parameters and biomarkers of CFTR function in 34 infants diagnosed with CF. Relationships with FIS were studied for indicators of pulmonary and gastrointestinal disease.Children with low FIS had higher levels of immunoreactive trypsinogen (p=0.030) and pancreatitis-associated protein (p=0.039), more often had pancreatic insufficiency (p<0.001), had more abnormalities on chest computed tomography (p=0.049), and had lower z-scores for maximal expiratory flow at functional residual capacity (p=0.033) when compared to children with high FIS values. FIS significantly correlated with sweat chloride concentration (SCC) and intestinal current measurement (ICM) (r= -0.82 and r=0.70, respectively; both p<0.001). Individual assessment of SCC, ICM and FIS suggested that FIS can help to classify individual disease severity.Thus, stratification by FIS identified subgroups that differed in pulmonary and gastrointestinal outcome parameters. FIS of intestinal organoids correlated well with established CFTR-dependent biomarkers such as SCC and ICM, and performed adequately at group and individual level in this proof-of-concept study.

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Section: Discussionmentioning

confidence: 99%

Stratifying infants with cystic fibrosis for disease severity using intestinal organoid swelling as a biomarker of CFTR function

et al. 2018

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“…Intestinal organoids can be disrupted to single cells to generate 2-D-monolayers on porous membranes for electrophysiological studies. FIS was shown to positively correlate with forskolin-induced current in subject-matched organoid-derived monolayers, supporting the notion that CFTR-dependent fluid secretion in rectal organoids reflects CFTR-dependent ion transport (Zomer-van Ommen et al, 2018)…”

Section: Forskolin-induced Swelling Assay As a Proxy For Cftr Functionmentioning

confidence: 52%

Human Primary Epithelial Cell Models: Promising Tools in the Era of Cystic Fibrosis Personalized Medicine

et al. 2018

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Cystic fibrosis (CF) is an inherited disorder where individual disease etiology and response to therapeutic intervention is impacted by CF transmembrane regulator (CFTR) mutations and other genetic modifiers. CFTR regulates multiple mechanisms in a diverse range of epithelial tissues. In this Review, we consolidate the latest updates in the development of primary epithelial cellular model systems relevant for CF. We discuss conventional two-dimensional (2-D) airway epithelial cell cultures, the backbone of in vitro cellular models to date, as well as improved expansion protocols to overcome finite supply of the cellular source. We highlight a range of strategies for establishment of three dimensional (3-D) airway and intestinal organoid models and evaluate the limitations and potential improvements in each system, focusing on their application in CF. The in vitro CFTR functional assays in patient-derived organoids allow for preclinical pharmacotherapy screening to identify responsive patients. It is likely that organoids will be an invaluable preclinical tool to unravel disease mechanisms, design novel treatments, and enable clinicians to provide personalized management for patients with CF.

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“…Forskolin-induced swelling (FIS) quantitatively correlates with CFTR function, as rapid FIS was observed in healthy control organoids, absent FIS in patients with CFTR null alleles, reduced FIS in milder CFTR mutations and strongly reduced FIS in organoids with severe CFTR mutations. This correlated with sweat chloride and intestinal current measurement as established biomarkers of CFTR function [31][32][33]. FIS is observed when CFTR function is restored in CF organoids with the addition of CFTR modulators and allows the assessment of individual drug responses [32].…”

Section: Theratyping Using Organoidsmentioning

confidence: 82%

Theratyping in cystic fibrosis

Crawford

Downey

2018

Current Opinion in Pulmonary Medicine

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Theratyping charts patients' clinical response to different treatments on an individual basis. This overcomes the limitations of genotype being used to predict response to individual therapies and includes all patients regardless of mutation. The use of organoids in high throughput screening allows numerous compounds to be tested on patient-specific tissue preclinically. This could lead to the extension of theratyping beyond CFTR modulators.

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Comparison of ex vivo and in vitro intestinal cystic fibrosis models to measure CFTR-dependent ion channel activity

Abstract: CFTR-dependent intestinal epithelial ion transport properties can be measured in rectal organoid-derived monolayers of subjects and correlate with donor-matched ICM and rectal organoid swelling.

Cited by 39 publications

References 34 publications

Stratifying infants with cystic fibrosis for disease severity using intestinal organoid swelling as a biomarker of CFTR function

Stratifying infants with cystic fibrosis for disease severity using intestinal organoid swelling as a biomarker of CFTR function

Human Primary Epithelial Cell Models: Promising Tools in the Era of Cystic Fibrosis Personalized Medicine

Theratyping in cystic fibrosis

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