Validation of Nasospheroids to Assay CFTR Functionality and Modulator Responses in Cystic Fibrosis

Overview

Journal Sci Rep

Specialty Science

Date 2021 Jul 31

PMID 34330959

Citations 5

Authors

Maite Calucho

Silvia Gartner

Paula Barranco

Paula Fernandez-Alvarez

Raquel Garcia Perez

Eduardo F Tizzano

Affiliations

Soon will be listed here.

Abstract

The availability of a simple, robust and non-invasive in vitro airway model would be useful to study the functionality of the cystic fibrosis transmembrane regulator (CFTR) protein and to personalize modulator therapy for cystic fibrosis (CF) patients. Our aim was to validate a CFTR functional study using nasospheroids, a patient-derived nasal cell 3D-culture. We performed live-cell experiments in nasospheroids obtained from wild-type individuals and CF patients with different genotypes and phenotypes. We extended the existing method and expanded the analysis to upgrade measurements of CFTR activity using forskolin-induced shrinking. We also tested modulator drugs in CF samples. Immobilizing suspended-nasospheroids provided a high number of samples for live-cell imaging. The diversity observed in basal sizes of nasospheroids did not affect the functional analysis of CFTR. Statistical analysis with our method was simple, making this protocol easy to reproduce. Moreover, we implemented the measurement of inner fluid reservoir areas to further differentiate CFTR functionality. In summary, this rapid methodology is helpful to analyse response to modulators in CF samples to allow individualized treatment for CF patients.

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References

Anderson J, Liu Z, Odom L, Kersh L, Guimbellot J . CFTR function and clinical response to modulators parallel nasal epithelial organoid swelling. Am J Physiol Lung Cell Mol Physiol. 2021; 321(1):L119-L129. PMC: 8321858. DOI: 10.1152/ajplung.00639.2020. View

Sachs N, Papaspyropoulos A, Zomer-van Ommen D, Heo I, Bottinger L, Klay D . Long-term expanding human airway organoids for disease modeling. EMBO J. 2019; 38(4). PMC: 6376275. DOI: 10.15252/embj.2018100300. View

Taylor-Cousar J, Munck A, McKone E, van der Ent C, Moeller A, Simard C . Tezacaftor-Ivacaftor in Patients with Cystic Fibrosis Homozygous for Phe508del. N Engl J Med. 2017; 377(21):2013-2023. DOI: 10.1056/NEJMoa1709846. View

McHugh D, Steele M, Valerio D, Miron A, Mann R, LePage D . A G542X cystic fibrosis mouse model for examining nonsense mutation directed therapies. PLoS One. 2018; 13(6):e0199573. PMC: 6010256. DOI: 10.1371/journal.pone.0199573. View

Lee R, Miller S, Mascenik T, Lewis C, Dang H, Boggs Z . Assessing Human Airway Epithelial Progenitor Cells for Cystic Fibrosis Cell Therapy. Am J Respir Cell Mol Biol. 2020; 63(3):374-385. PMC: 7462339. DOI: 10.1165/rcmb.2019-0384OC. View

Brewington J, Filbrandt E, LaRosa 3rd F, Moncivaiz J, Ostmann A, Strecker L . Brushed nasal epithelial cells are a surrogate for bronchial epithelial CFTR studies. JCI Insight. 2018; 3(13). PMC: 6124537. DOI: 10.1172/jci.insight.99385. View

Taylor-Cousar J, Jain M, Barto T, Haddad T, Atkinson J, Tian S . Lumacaftor/ivacaftor in patients with cystic fibrosis and advanced lung disease homozygous for F508del-CFTR. J Cyst Fibros. 2017; 17(2):228-235. DOI: 10.1016/j.jcf.2017.09.012. View

Brewington J, Filbrandt E, LaRosa 3rd F, Moncivaiz J, Ostmann A, Strecker L . Generation of Human Nasal Epithelial Cell Spheroids for Individualized Cystic Fibrosis Transmembrane Conductance Regulator Study. J Vis Exp. 2018; (134). PMC: 5933499. DOI: 10.3791/57492. View

Bland J, Altman D . Transformations, means, and confidence intervals. BMJ. 1996; 312(7038):1079. PMC: 2350916. DOI: 10.1136/bmj.312.7038.1079. View

10.

Awatade N, Wong S, Hewson C, Fawcett L, Kicic A, Jaffe A . Human Primary Epithelial Cell Models: Promising Tools in the Era of Cystic Fibrosis Personalized Medicine. Front Pharmacol. 2018; 9:1429. PMC: 6293199. DOI: 10.3389/fphar.2018.01429. View

11.

Kerem B, Rommens J, Buchanan J, Markiewicz D, Cox T, Chakravarti A . Identification of the cystic fibrosis gene: genetic analysis. Science. 1989; 245(4922):1073-80. DOI: 10.1126/science.2570460. View

12.

Brewington J, Filbrandt E, LaRosa 3rd F, Ostmann A, Strecker L, Szczesniak R . Detection of CFTR function and modulation in primary human nasal cell spheroids. J Cyst Fibros. 2017; 17(1):26-33. PMC: 5868354. DOI: 10.1016/j.jcf.2017.06.010. View

13.

Liu Z, Anderson J, Deng L, Mackay S, Bailey J, Kersh L . Human Nasal Epithelial Organoids for Therapeutic Development in Cystic Fibrosis. Genes (Basel). 2020; 11(6). PMC: 7349680. DOI: 10.3390/genes11060603. View

14.

Wainwright C, Elborn J, Ramsey B, Marigowda G, Huang X, Cipolli M . Lumacaftor-Ivacaftor in Patients with Cystic Fibrosis Homozygous for Phe508del CFTR. N Engl J Med. 2015; 373(3):220-31. PMC: 4764353. DOI: 10.1056/NEJMoa1409547. View

15.

Boyle M, Bell S, Konstan M, McColley S, Rowe S, Rietschel E . A CFTR corrector (lumacaftor) and a CFTR potentiator (ivacaftor) for treatment of patients with cystic fibrosis who have a phe508del CFTR mutation: a phase 2 randomised controlled trial. Lancet Respir Med. 2014; 2(7):527-38. DOI: 10.1016/S2213-2600(14)70132-8. View

16.

Egan M . Cystic fibrosis transmembrane conductance receptor modulator therapy in cystic fibrosis, an update. Curr Opin Pediatr. 2020; 32(3):384-388. DOI: 10.1097/MOP.0000000000000892. View

17.

Crawford D, Mullenders J, Pott J, Boj S, Landskroner-Eiger S, Goddeeris M . Targeting G542X CFTR nonsense alleles with ELX-02 restores CFTR function in human-derived intestinal organoids. J Cyst Fibros. 2021; 20(3):436-442. DOI: 10.1016/j.jcf.2021.01.009. View

18.

Clancy J, Cotton C, Donaldson S, Solomon G, VanDevanter D, Boyle M . CFTR modulator theratyping: Current status, gaps and future directions. J Cyst Fibros. 2018; 18(1):22-34. PMC: 6301143. DOI: 10.1016/j.jcf.2018.05.004. View

19.

Van Goor F, Hadida S, Grootenhuis P, Burton B, Cao D, Neuberger T . Rescue of CF airway epithelial cell function in vitro by a CFTR potentiator, VX-770. Proc Natl Acad Sci U S A. 2009; 106(44):18825-30. PMC: 2773991. DOI: 10.1073/pnas.0904709106. View

20.

Konstan M, McKone E, Moss R, Marigowda G, Tian S, Waltz D . Assessment of safety and efficacy of long-term treatment with combination lumacaftor and ivacaftor therapy in patients with cystic fibrosis homozygous for the F508del-CFTR mutation (PROGRESS): a phase 3, extension study. Lancet Respir Med. 2016; 5(2):107-118. DOI: 10.1016/S2213-2600(16)30427-1. View