Nasal polyp fibroblasts modulate epithelial characteristics via Wnt signaling

Alex Dobzanski, Syed Muaz Khalil, Andrew P Lane

Research output: Contribution to journalArticle

Abstract

Background: While essential to the normal differentiation of ciliated airway epithelial cells, upregulated Wnt signaling in chronic rhinosinusitis with nasal polyps (CRSwNP) has been proposed to result in abnormal epithelial morphology and dysfunctional mucociliary clearance. The mechanism of epithelial Wnt signaling dysregulation in CRSwNP is unknown, and importantly cellular sources of Wnt ligands in CRSwNP have not yet been investigated. Methods: Human sinonasal epithelial cells (hSNECs) and human sinonasal fibroblasts (hSNFs) were collected from 34 human subjects (25 control and 9 CRSwNP) and differentiated as primary air-liquid interface (ALI) and organoid co-cultures. hSNECs were isolated to the apical compartment of the transwell and hSNFs were isolated to the basolateral compartment. After 21 days of ALI culture, ciliary expression and sinonasal epithelial morphology were examined by immunohistochemistry (IHC) and quantitative real-time polymerase chain reaction (qRT-PCR). An organoid model was used to evaluate proliferation of basal cells in presence of hSNFs. Results: Epithelial cells co-cultured with CRSwNP-hSNFs revealed significantly decreased ciliated cells, altered epithelial cell morphology, and increased colony forming efficiency compared to epithelial cells co-cultured with control-hSNFs. CRSwNP-hSNFs showed significantly higher messenger RNA (mRNA) expression of canonical WNT3A. A Wnt agonist, CHIR99021, replicated CRSwNP-hSNF co-cultures, and treatment with the Wnt inhibitor IWP2 prevented abnormal morphologies. Conclusion: These results suggest that abnormal interactions between epithelial cells and fibroblasts may contribute to CRSwNP pathogenesis and supports the concept that dysregulated Wnt signaling contributes impairment to epithelial function in CRSwNP.

Original languageEnglish (US)
JournalInternational Forum of Allergy and Rhinology
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Nasal Polyps
Fibroblasts
Epithelial Cells
Organoids
Coculture Techniques
Air
Mucociliary Clearance
Real-Time Polymerase Chain Reaction
Immunohistochemistry
Cell Proliferation
Ligands

Keywords

  • barrier
  • chronic rhinosinusitis with nasal polyps
  • ciliation
  • epithelial-mesenchymal transition
  • organoid
  • Wnt

ASJC Scopus subject areas

  • Immunology and Allergy
  • Otorhinolaryngology

Cite this

Nasal polyp fibroblasts modulate epithelial characteristics via Wnt signaling. / Dobzanski, Alex; Khalil, Syed Muaz; Lane, Andrew P.

In: International Forum of Allergy and Rhinology, 01.01.2018.

Research output: Contribution to journalArticle

@article{929560c3bebe4d468ccb705957d8f3f8,
title = "Nasal polyp fibroblasts modulate epithelial characteristics via Wnt signaling",
abstract = "Background: While essential to the normal differentiation of ciliated airway epithelial cells, upregulated Wnt signaling in chronic rhinosinusitis with nasal polyps (CRSwNP) has been proposed to result in abnormal epithelial morphology and dysfunctional mucociliary clearance. The mechanism of epithelial Wnt signaling dysregulation in CRSwNP is unknown, and importantly cellular sources of Wnt ligands in CRSwNP have not yet been investigated. Methods: Human sinonasal epithelial cells (hSNECs) and human sinonasal fibroblasts (hSNFs) were collected from 34 human subjects (25 control and 9 CRSwNP) and differentiated as primary air-liquid interface (ALI) and organoid co-cultures. hSNECs were isolated to the apical compartment of the transwell and hSNFs were isolated to the basolateral compartment. After 21 days of ALI culture, ciliary expression and sinonasal epithelial morphology were examined by immunohistochemistry (IHC) and quantitative real-time polymerase chain reaction (qRT-PCR). An organoid model was used to evaluate proliferation of basal cells in presence of hSNFs. Results: Epithelial cells co-cultured with CRSwNP-hSNFs revealed significantly decreased ciliated cells, altered epithelial cell morphology, and increased colony forming efficiency compared to epithelial cells co-cultured with control-hSNFs. CRSwNP-hSNFs showed significantly higher messenger RNA (mRNA) expression of canonical WNT3A. A Wnt agonist, CHIR99021, replicated CRSwNP-hSNF co-cultures, and treatment with the Wnt inhibitor IWP2 prevented abnormal morphologies. Conclusion: These results suggest that abnormal interactions between epithelial cells and fibroblasts may contribute to CRSwNP pathogenesis and supports the concept that dysregulated Wnt signaling contributes impairment to epithelial function in CRSwNP.",
keywords = "barrier, chronic rhinosinusitis with nasal polyps, ciliation, epithelial-mesenchymal transition, organoid, Wnt",
author = "Alex Dobzanski and Khalil, {Syed Muaz} and Lane, {Andrew P}",
year = "2018",
month = "1",
day = "1",
doi = "10.1002/alr.22199",
language = "English (US)",
journal = "International Forum of Allergy and Rhinology",
issn = "2042-6976",
publisher = "Wiley-Blackwell",

}

TY - JOUR

T1 - Nasal polyp fibroblasts modulate epithelial characteristics via Wnt signaling

AU - Dobzanski, Alex

AU - Khalil, Syed Muaz

AU - Lane, Andrew P

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Background: While essential to the normal differentiation of ciliated airway epithelial cells, upregulated Wnt signaling in chronic rhinosinusitis with nasal polyps (CRSwNP) has been proposed to result in abnormal epithelial morphology and dysfunctional mucociliary clearance. The mechanism of epithelial Wnt signaling dysregulation in CRSwNP is unknown, and importantly cellular sources of Wnt ligands in CRSwNP have not yet been investigated. Methods: Human sinonasal epithelial cells (hSNECs) and human sinonasal fibroblasts (hSNFs) were collected from 34 human subjects (25 control and 9 CRSwNP) and differentiated as primary air-liquid interface (ALI) and organoid co-cultures. hSNECs were isolated to the apical compartment of the transwell and hSNFs were isolated to the basolateral compartment. After 21 days of ALI culture, ciliary expression and sinonasal epithelial morphology were examined by immunohistochemistry (IHC) and quantitative real-time polymerase chain reaction (qRT-PCR). An organoid model was used to evaluate proliferation of basal cells in presence of hSNFs. Results: Epithelial cells co-cultured with CRSwNP-hSNFs revealed significantly decreased ciliated cells, altered epithelial cell morphology, and increased colony forming efficiency compared to epithelial cells co-cultured with control-hSNFs. CRSwNP-hSNFs showed significantly higher messenger RNA (mRNA) expression of canonical WNT3A. A Wnt agonist, CHIR99021, replicated CRSwNP-hSNF co-cultures, and treatment with the Wnt inhibitor IWP2 prevented abnormal morphologies. Conclusion: These results suggest that abnormal interactions between epithelial cells and fibroblasts may contribute to CRSwNP pathogenesis and supports the concept that dysregulated Wnt signaling contributes impairment to epithelial function in CRSwNP.

AB - Background: While essential to the normal differentiation of ciliated airway epithelial cells, upregulated Wnt signaling in chronic rhinosinusitis with nasal polyps (CRSwNP) has been proposed to result in abnormal epithelial morphology and dysfunctional mucociliary clearance. The mechanism of epithelial Wnt signaling dysregulation in CRSwNP is unknown, and importantly cellular sources of Wnt ligands in CRSwNP have not yet been investigated. Methods: Human sinonasal epithelial cells (hSNECs) and human sinonasal fibroblasts (hSNFs) were collected from 34 human subjects (25 control and 9 CRSwNP) and differentiated as primary air-liquid interface (ALI) and organoid co-cultures. hSNECs were isolated to the apical compartment of the transwell and hSNFs were isolated to the basolateral compartment. After 21 days of ALI culture, ciliary expression and sinonasal epithelial morphology were examined by immunohistochemistry (IHC) and quantitative real-time polymerase chain reaction (qRT-PCR). An organoid model was used to evaluate proliferation of basal cells in presence of hSNFs. Results: Epithelial cells co-cultured with CRSwNP-hSNFs revealed significantly decreased ciliated cells, altered epithelial cell morphology, and increased colony forming efficiency compared to epithelial cells co-cultured with control-hSNFs. CRSwNP-hSNFs showed significantly higher messenger RNA (mRNA) expression of canonical WNT3A. A Wnt agonist, CHIR99021, replicated CRSwNP-hSNF co-cultures, and treatment with the Wnt inhibitor IWP2 prevented abnormal morphologies. Conclusion: These results suggest that abnormal interactions between epithelial cells and fibroblasts may contribute to CRSwNP pathogenesis and supports the concept that dysregulated Wnt signaling contributes impairment to epithelial function in CRSwNP.

KW - barrier

KW - chronic rhinosinusitis with nasal polyps

KW - ciliation

KW - epithelial-mesenchymal transition

KW - organoid

KW - Wnt

UR - http://www.scopus.com/inward/record.url?scp=85052405507&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85052405507&partnerID=8YFLogxK

U2 - 10.1002/alr.22199

DO - 10.1002/alr.22199

M3 - Article

C2 - 30118173

AN - SCOPUS:85052405507

JO - International Forum of Allergy and Rhinology

JF - International Forum of Allergy and Rhinology

SN - 2042-6976

ER -