PMID- 37565709
OWN - NLM
STAT- MEDLINE
DCOM- 20231226
LR  - 20240207
IS  - 1531-4995 (Electronic)
IS  - 0023-852X (Print)
IS  - 0023-852X (Linking)
VI  - 134
IP  - 1
DP  - 2024 Jan
TI  - Dysfunctional Epithelial Barrier Is Characterized by Reduced E-Cadherin in 
      Idiopathic Subglottic Stenosis.
PG  - 374-381
LID - 10.1002/lary.30951 [doi]
AB  - OBJECTIVES: To aim of the study was to characterize the molecular profile and 
      functional phenotype of idiopathic subglottic stenosis (iSGS)-scar epithelium. 
      METHODS: Human tracheal biopsies from iSGS scar (n = 6) and matched non-scar 
      (n = 6) regions were analyzed using single-cell RNA sequencing (scRNA-seq). 
      Separate specimens were used for epithelial cell expansion in vitro to assess 
      average growth rate and functional capabilities using transepithelial-electrical 
      resistance (TEER), fluorescein isothiocyanate-dextran flux permeability assay, 
      ciliary coverage, and cilia beating frequency (CBF). Finally, epithelial tight 
      junction protein expression of cultured cells was quantified using immunoblot 
      assay (n = 4) and immunofluorescence (n = 6). RESULTS: scRNA-seq analysis 
      revealed a decrease in goblet, ciliated, and basal epithelial cells in the scar 
      iSGS cohort. Furthermore, mRNA expression of proteins E-cadherin, claudin-3, 
      claudin-10, occludin, TJP1, and TJP2 was also reduced (p < 0.001) in scar 
      epithelium. Functional assays demonstrated a decrease in TEER (paired 95% 
      confidence interval [CI], 195.68-890.83 Omega x cm(2) , p < 0.05), an increase in 
      permeability (paired 95% CI, -6116.00 to -1401.99 RFU, p < 0.05), and reduced 
      epithelial coverage (paired 95% CI, 0.1814-1.766, fold change p < 0.05) in 
      iSGS-scar epithelium relative to normal controls. No difference in growth rate 
      (p < 0.05) or CBF was found (paired 95% CI, -2.118 to 3.820 Hz, p > 0.05). 
      Immunoblot assay (paired 95% CI, 0.0367-0.605, p < 0.05) and immunofluorescence 
      (paired 95% CI, 13.748-59.191 mean grey value, p < 0.05) revealed E-cadherin 
      reduction in iSGS-scar epithelium. CONCLUSION: iSGS-scar epithelium has a 
      dysfunctional barrier and reduced structural protein expression. These results 
      are consistent with dysfunctional epithelium seen in other airway pathology. 
      Further studies are warranted to delineate the causality of epithelial 
      dysfunction on the downstream fibroinflammatory cascade in iSGS. LEVEL OF 
      EVIDENCE: NA Laryngoscope, 134:374-381, 2024.
CI  - (c) 2023 The American Laryngological, Rhinological and Otological Society, Inc.
FAU - Berges, Alexandra J
AU  - Berges AJ
AUID- ORCID: 0000-0002-0890-6291
AD  - Johns Hopkins Department of Otolaryngology-Head and Neck Surgery, Baltimore, 
      Maryland, U.S.A.
FAU - Ospino, Rafael
AU  - Ospino R
AUID- ORCID: 0000-0002-8501-1934
AD  - Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A.
FAU - Mafla, Laura
AU  - Mafla L
AD  - Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A.
FAU - Collins, Samuel
AU  - Collins S
AD  - Johns Hopkins Department of Otolaryngology-Head and Neck Surgery, Baltimore, 
      Maryland, U.S.A.
FAU - Chan-Li, Yee
AU  - Chan-Li Y
AUID- ORCID: 0000-0001-9781-0728
AD  - Johns Hopkins Department of Otolaryngology-Head and Neck Surgery, Baltimore, 
      Maryland, U.S.A.
FAU - Ghosh, Baishakhi
AU  - Ghosh B
AD  - Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, U.S.A.
FAU - Sidhaye, Venkataramana
AU  - Sidhaye V
AD  - Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, U.S.A.
AD  - Johns Hopkins Division of Pulmonary and Critical Care Medicine, Baltimore, 
      Maryland, U.S.A.
FAU - Lina, Ioan
AU  - Lina I
AUID- ORCID: 0000-0002-8619-7312
AD  - Johns Hopkins Department of Otolaryngology-Head and Neck Surgery, Baltimore, 
      Maryland, U.S.A.
FAU - Hillel, Alexander T
AU  - Hillel AT
AUID- ORCID: 0000-0001-8471-5449
AD  - Johns Hopkins Department of Otolaryngology-Head and Neck Surgery, Baltimore, 
      Maryland, U.S.A.
LA  - eng
GR  - R01 DC018567/DC/NIDCD NIH HHS/United States
GR  - 1R01DC018567/DC/NIDCD NIH HHS/United States
GR  - R56 DC020736/DC/NIDCD NIH HHS/United States
GR  - R01 HL124099/HL/NHLBI NIH HHS/United States
GR  - R01 HL151107/HL/NHLBI NIH HHS/United States
GR  - R01HL124099/HL/NHLBI NIH HHS/United States
GR  - R56DC020736/DC/NIDCD NIH HHS/United States
GR  - HLR01HL151107/HL/NHLBI NIH HHS/United States
PT  - Journal Article
DEP - 20230810
PL  - United States
TA  - Laryngoscope
JT  - The Laryngoscope
JID - 8607378
RN  - 0 (Cadherins)
SB  - IM
MH  - Humans
MH  - *Cadherins/metabolism
MH  - *Cicatrix/metabolism
MH  - Constriction, Pathologic
MH  - Epithelium/metabolism
MH  - Epithelial Cells/metabolism
MH  - Permeability
PMC - PMC10842128
MID - NIHMS1940101
OTO - NOTNLM
OT  - E-cadherin
OT  - epithelial barrier dysfunction
OT  - epithelium
OT  - idiopathic subglottic stenosis
OT  - occludin
OT  - single-cell analysis
COIS- Dr. Hillel had a sponsored research agreement with Medtronic to investigate 
      tracheostomy tube injury to the trachea. The remaining authors have no other 
      funding, financial relationships, or conflicts of interest to disclose.
EDAT- 2023/08/11 12:42
MHDA- 2023/12/26 06:41
PMCR- 2025/01/01
CRDT- 2023/08/11 08:34
PHST- 2023/07/11 00:00 [revised]
PHST- 2023/04/06 00:00 [received]
PHST- 2023/07/18 00:00 [accepted]
PHST- 2025/01/01 00:00 [pmc-release]
PHST- 2023/12/26 06:41 [medline]
PHST- 2023/08/11 12:42 [pubmed]
PHST- 2023/08/11 08:34 [entrez]
AID - 10.1002/lary.30951 [doi]
PST - ppublish
SO  - Laryngoscope. 2024 Jan;134(1):374-381. doi: 10.1002/lary.30951. Epub 2023 Aug 10.