PMID- 36524603
OWN - NLM
STAT- MEDLINE
DCOM- 20230811
LR  - 20231219
IS  - 1531-4995 (Electronic)
IS  - 0023-852X (Print)
IS  - 0023-852X (Linking)
VI  - 133
IP  - 9
DP  - 2023 Sep
TI  - Characterizing the Macrophage Population in Patients With Idiopathic Subglottic 
      Stenosis.
PG  - 2308-2316
LID - 10.1002/lary.30524 [doi]
AB  - OBJECTIVES: Idiopathic subglottic stenosis (iSGS) is characterized by progressive 
      fibrosis and subglottic luminal narrowing. Currently, immune characterization has 
      focused on T-cells; however, macrophages remain largely unexplored. The goals of 
      this study are to characterize the transcriptome of iSGS macrophages and the 
      fibrogenic nature of identifed biomarkers. STUDY DESIGN: Bioinformatics and in 
      vitro. METHODS: Human tracheal biopsies from iSGS scar (n = 4), and matched 
      non-scar (n = 4) regions were analyzed using single-cell RNA-seq (scRNA-seq). 
      Immunofluorescence (IF) was performed on rapidly processed autopsies (RPA) and 
      iSGS tracheal resections (n = 4) to co-localize S100A8/9 and CD11b. Collagen 
      gene/protein expression was assessed in iSGS fibroblasts (n = 4) treated with 
      protein S100A8/9 (1000 ng/ml). Macrophages were subclustered to identify distinct 
      subpopulations. RESULTS: scRNA-seq analysis revealed S100A8/S100A9 (fold change 
      (FC) = 4.1/1.88, p < 0.001) as top differentially expressed genes in iSGS 
      macrophages. IF exhibited increased CD11b+/S100A8/9+ cells in tracheal samples of 
      iSGS versus RPA (26.75% +/- 7.08 vs. 0.594% +/- 0.974, n = 4, p = 0.029). iSGS 
      fibroblasts treated with S100A8/9 demonstrated increased gene expression of 
      COL1A1 (FC = 2.30 +/- 0.45, p = 0.03, n = 4) and COL3A1 (FC = 2.44 +/- 0.40, 
      p = 0.03, n = 4). COL1A1 protein assays revealed an increase in the experimental 
      group, albeit not significant, (p = 0.12, n = 4). Finally, macrophage sub 
      clustering revealed one subpopulation as a predominant source of S100A8/S100A9 
      expression (FC = 7.94/5.47, p < 0.001). CONCLUSIONS: S100A8/9 is a key biomarker 
      in iSGS macrophages. Although S100A8/9 demonstrates profibrotic nature in vitro, 
      the role of S100A8/9+ macrophages in vivo warrants further investigation. LEVEL 
      OF EVIDENCE: NA Laryngoscope, 133:2308-2316, 2023.
CI  - (c) 2022 The American Laryngological, Rhinological and Otological Society, Inc.
FAU - Ospino, Rafael
AU  - Ospino R
AUID- ORCID: 0000-0002-8501-1934
AD  - Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA.
FAU - Berges, Alexandra
AU  - Berges A
AUID- ORCID: 0000-0002-0890-6291
AD  - Department of Otolaryngology-Head & Neck Surgery, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland, USA.
FAU - Mafla, Laura
AU  - Mafla L
AD  - Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA.
FAU - Collins, Samuel
AU  - Collins S
AD  - Department of Otolaryngology-Head & Neck Surgery, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland, USA.
FAU - Li, Yee Chan
AU  - Li YC
AUID- ORCID: 0000-0002-5357-8776
AD  - Department of Otolaryngology-Head & Neck Surgery, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland, USA.
FAU - Lina, Ioan
AU  - Lina I
AUID- ORCID: 0000-0002-8619-7312
AD  - Department of Otolaryngology-Head & Neck Surgery, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland, USA.
FAU - Gelbard, Alexander
AU  - Gelbard A
AUID- ORCID: 0000-0003-0078-1305
AD  - Department of Otolaryngology-Head & Neck Surgery, Vanderbilt University School of 
      Medicine, Nashville, Tennessee, USA.
FAU - Hillel, Alexander T
AU  - Hillel AT
AUID- ORCID: 0000-0001-8471-5449
AD  - Department of Otolaryngology-Head & Neck Surgery, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland, USA.
FAU - Motz, Kevin
AU  - Motz K
AUID- ORCID: 0000-0002-9255-7265
AD  - Department of Otolaryngology-Head & Neck Surgery, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland, USA.
LA  - eng
GR  - R01 DC018567/DC/NIDCD NIH HHS/United States
GR  - R21 DC017225/DC/NIDCD NIH HHS/United States
GR  - K23 DC020988/DC/NIDCD NIH HHS/United States
GR  - R01 HL146401/HL/NHLBI NIH HHS/United States
GR  - K23 DC014082/DC/NIDCD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20221216
PL  - United States
TA  - Laryngoscope
JT  - The Laryngoscope
JID - 8607378
RN  - 0 (Calgranulin A)
RN  - 0 (Calgranulin B)
SB  - IM
MH  - Humans
MH  - Constriction, Pathologic
MH  - *Laryngostenosis/pathology
MH  - Macrophages/metabolism
MH  - Calgranulin A/genetics/metabolism
MH  - Calgranulin B/genetics/metabolism
PMC - PMC10272290
MID - NIHMS1859935
OTO - NOTNLM
OT  - S100A8/9
OT  - idiopathic subglottic stenosis
OT  - macrophages
OT  - single cell analysis
COIS- Conflict of Interest Statement: The authors have no funding, financial 
      relationships, or conflicts of interest to disclose.
EDAT- 2022/12/17 06:00
MHDA- 2023/08/11 06:43
PMCR- 2024/09/01
CRDT- 2022/12/16 06:23
PHST- 2022/09/19 00:00 [revised]
PHST- 2022/04/07 00:00 [received]
PHST- 2022/11/20 00:00 [accepted]
PHST- 2024/09/01 00:00 [pmc-release]
PHST- 2023/08/11 06:43 [medline]
PHST- 2022/12/17 06:00 [pubmed]
PHST- 2022/12/16 06:23 [entrez]
AID - 10.1002/lary.30524 [doi]
PST - ppublish
SO  - Laryngoscope. 2023 Sep;133(9):2308-2316. doi: 10.1002/lary.30524. Epub 2022 Dec 
      16.