Assemblies of JAG1 and JAG2 determine tracheobronchial cell fate in mucosecretory lung disease
Susan D. Reynolds, Cynthia L. Hill, Alfahdah Alsudayri, Scott W. Lallier, Saranga Wijeratne, Zheng Hong Tan, Tendy Chiang, Estelle Cormet-Boyaka
Susan D. Reynolds, Cynthia L. Hill, Alfahdah Alsudayri, Scott W. Lallier, Saranga Wijeratne, Zheng Hong Tan, Tendy Chiang, Estelle Cormet-Boyaka
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Research Article Cell biology Stem cells

Assemblies of JAG1 and JAG2 determine tracheobronchial cell fate in mucosecretory lung disease

Abstract

Mucosecretory lung disease compromises airway epithelial function and is characterized by goblet cell hyperplasia and ciliated cell hypoplasia. Goblet and ciliated cell types are derived from tracheobronchial stem/progenitor cells via a Notch-dependent mechanism. Although specific arrays of Notch receptors regulate cell fate determination, the function of the ligands Jagged1 (JAG1) and JAG2 is unclear. This study examined JAG1 and JAG2 function using human air-liquid-interface cultures that were treated with γ-secretase complex (GSC) inhibitors, neutralizing peptides/antibodies, or WNT/β-catenin pathway antagonists/agonists. These experiments revealed that JAG1 and JAG2 regulated cell fate determination in the tracheobronchial epithelium; however, their roles did not adhere to simple necessity and sufficiency rules. Biochemical studies indicated that JAG1 and JAG2 underwent posttranslational modifications that resulted in generation of a JAG1 C-terminal peptide and regulated the abundance of full-length JAG2 on the cell surface. GSC and glycogen synthase kinase 3 were implicated in these posttranslational events, but WNT agonist/antagonist studies and RNA-Seq indicated a WNT-independent mechanism. Collectively, these data suggest that posttranslational modifications create distinct assemblies of JAG1 and JAG2, which regulate Notch signal strength and determine the fate of tracheobronchial stem/progenitor cells.

Authors

Susan D. Reynolds, Cynthia L. Hill, Alfahdah Alsudayri, Scott W. Lallier, Saranga Wijeratne, Zheng Hong Tan, Tendy Chiang, Estelle Cormet-Boyaka

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Figure 7

scRNA-Seq analysis of human bronchial basal cell differentiation.

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scRNA-Seq analysis of human bronchial basal cell differentiation. Human ...
Human bronchial basal cells were differentiated in ALI cultures using H&H medium, treated with vehicle (DMSO) or 10 μM CHIR on day 4 and day 6, and harvested for scRNA-Seq analysis on day 8. (A–D) Uniform Manifold Approximation and Projection (UMAP) analysis. (A) Cluster identification color code. UMAP analysis of cells that were treated with (B) vehicle or CHIR, (C) vehicle, or (D) CHIR. (E) Dot plot of the marker genes used to identify the clusters. For each dot, the color gradient (blue to red) indicates the mean marker expression, and the size indicates the percentage of cells expressing the marker. See Supplemental Table 1 for the complete gene list. Cluster identification by cell phenotype or state: 0, secretory lineage, presecretory primed; 1, classical basal cells; 2, differentiating basal cells; 3, secretory lineage, secretory primed cell; 4, secretory lineage, presecretory primed; 5, ciliated lineage, multiciliated cells; 6, self-renewing cells; 7, motile/squamous cells; 8, self-renewing cells; 9, motile/squamous cells; 10, differentiating basal cells; 11, ciliated lineage; 12, secretory lineage, mature secretory cells; 13, ciliated lineage, bristle/deuterostome cells. (F–I) Inference of basal, secretory, squamous, and ciliated cell lineages by Monocle 3. (F) Pseudotime is represented by the purple (start) to yellow (end) gradient and is (G) based on an aggregate of the entire experiment, (H) vehicle-treated and (I) CHIR-treated cells. The red arrows indicate the assigned root node, and the differentiation trajectory is indicated by the black line.