Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Aug 15;210(4):517-521.
doi: 10.1164/rccm.202311-2111LE.

MUC5B Idiopathic Pulmonary Fibrosis Risk Variant Promotes a Mucosecretory Phenotype and Loss of Small Airway Secretory Cells

Jonathan S Kurche  1   2 , Carlyne D Cool  3   4 , Rachel Z Blumhagen  5 , Evgenia Dobrinskikh  6 , David Heinz  4 , Jeremy A Herrera  2 , Ivana V Yang  2   7 , David A Schwartz  8   2
Affiliations

MUC5B Idiopathic Pulmonary Fibrosis Risk Variant Promotes a Mucosecretory Phenotype and Loss of Small Airway Secretory Cells

Jonathan S Kurche et al. Am J Respir Crit Care Med. .
No abstract available

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
The MUC5B promoter variant is associated with reduced distal lung differentiation markers in the small airway. (A) Volcano plot of significant differentially expressed genes in the small airway, based on a dominant model for MUC5B promoter genotype. MUC5B non–risk variant (G) is associated with expression of SCGB3A2 and NKX2-1, whereas the risk variant (T) is associated with MUC5B and MUC5AC expression, among others (n = 46). Shown are results of mixed effects modeling adjusting for disease (idiopathic pulmonary fibrosis [IPF] or unaffected), batch, region of interest (ROI) area, and subject identity as a random effect to correct for multiple samples from the same specimen. (B) MUC5B non–risk variant (G) is associated with ERAP1, ANXA4, HGSNAT, and CNGA4 and nonsignificantly associated with MUC5B and MUC5AC expression in control tissues (n = 11). Shown are results of mixed effects modeling of spatial transcriptional profiles of control airways adjusting for batch and ROI area, with subject identifier as a random effect. (C) MUC5B genotype effects in IPF airways reveal significant effects on MUC5B expression among others (n = 35). Shown are results of mixed effects modeling of spatial transcriptional profiles of IPF airways adjusting for batch and ROI area, with subject identifier as a random effect. In all cases, the x-axis represents fold change, and the y-axis represents −log10 Benjamini-Hochberg adjusted FDR; lines at 1.5-fold change (0.67 log2 fold change) and adjusted P = 0.05. FDR = false discovery rate.
Figure 2.
Figure 2.
Fewer small airway secretory cells (SASCs) present in MUC5B variant carriers. Presence of the MUC5B idiopathic pulmonary fibrosis (IPF) risk variant (MUC5B common allele [G] or MUC5B variant [T], based on a dominant genetic model) is associated with a reduced proportion of SFTPB+, SCGB3A2+ double-positive SASCs relative to total airway nucleated cells. (A) Immunofluorescence staining of formalin-fixed, paraffin-embedded distal airways demonstrating MUC5B (green, HPA008246, Sigma-Aldrich, 1:200), SFTPB (magenta, MAI-204, Invitrogen, 1:200), and SCGB3A2 (yellow, AF3545, R&D Systems, 1:200) staining across representative specimens of genotype and condition. We used citrate-buffered antigen retrieval and standard immunofluorescence approaches (17). For each airway, 20× immunofluorescence images were obtained using a Keyence BZ-X800 inverted microscope. Scale bars, 100 μm. (B–D) Proportion of SFTPB+ SCGB3A2+ double-positive SASCs within the airway as a fraction of total DAPI+ nuclei based on (B) disease, (C) genotype (MUC5B common allele (G) or MUC5B variant [T]), and (D) disease and genotype. Images were exported as TIF files into the Fiji implementation of ImageJ (18), cropped to exclude alveolar tissues with the freehand tool, thresholded to eliminate background, converted to masks, and quantified by area and number of fluorescent foci. Measurements were normalized according to the numbers of DAPI+ nuclei present in the selection, and the percentage of SFTPB+, SCGB3A2+ double-positive SASCs out of total DAPI+ nuclei was recorded. Data were exported to GraphPad Prism (GraphPad Software), where differences in cell percentages among IPF and controls or nonvariant (G) and variant (T) MUC5B alleles were analyzed for significance according to Mann-Whitney U test (B and C). For (D), the Kruskal-Wallis method was used, with secondary pairwise comparisons made according to Dunn’s test and significance set to a nominal P value of ⩽0.05. Bars are set to mean values.
See this image and copyright information in PMC

References

    1. Seibold MA, Wise AL, Speer MC, Steele MP, Brown KK, Loyd JE, et al. A common MUC5B promoter polymorphism and pulmonary fibrosis. N Engl J Med . 2011;364:1503–1512. - PMC - PubMed
    1. Nakano Y, Yang IV, Walts AD, Watson AM, Helling BA, Fletcher AA, et al. MUC5B promoter variant rs35705950 affects MUC5B expression in the distal airways in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med . 2016;193:464–466. - PMC - PubMed
    1. Shih AR, Nitiwarangkul C, Little BP, Roop BW, Nandy S, Szabari MV, et al. Practical application and validation of the 2018 ATS/ERS/JRS/ALAT and Fleischner Society guidelines for the diagnosis of idiopathic pulmonary fibrosis. Respir Res . 2021;22:124. - PMC - PubMed
    1. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol . 2014;15:550. - PMC - PubMed
    1. Vestal BE, Wynn E, Moore CM. lmerSeq: an R package for analyzing transformed RNA-Seq data with linear mixed effects models. BMC Bioinformatics . 2022;23:489. - PMC - PubMed

LinkOut - more resources