Trefoil Factor Family 3 Peptide Promotes Human Airway Epithelial Ciliated Cell Differentiation

Abstract

Human airway surface epithelium is frequently damaged by inhaled factors (viruses, bacteria, xenobiotic substances) as well as by inflammatory mediators that contribute to the shedding of surface epithelial cells. To regain its protective function, the epithelium must rapidly repair and redifferentiate. The Trefoil Factor Family (TFF) peptides are secretory products of many mucous cells. TFF3, the major TFF in the airways, is able to enhance airway epithelial cell migration, but the role of this protein in differentiation has not been defined. To identify the specific role of TFF3 in the differentiation of the human airway surface epithelium, we analyzed the temporal expression pattern of TFF3, MUC5AC, and MUC5B mucins (goblet cells) and ciliated cell markers beta-tubulin (cilia) and FOXJ1 (ciliogenesis) during human airway epithelial regeneration using in vivo humanized airway xenograft and in vitro air-liquid interface (ALI) culture models. We observed that TFF3, MUC5AC, MUC5B, and ciliated cell markers were expressed in well-differentiated airway epithelium. The addition of exogenous recombinant human TFF3 to epithelial cell cultures before the initiation of differentiation resulted in no change in MUC5AC or cytokeratin 13 (CK13, basal cell marker)-positive cells, but induced an increase in the number of FOXJ1-positive cells and in the number of beta-tubulin-positive ciliated cells (P < 0.05). Furthermore, this effect on ciliated cell differentiation could be reversed by specific epidermal growth factor (EGF) receptor (EGF-R) inhibition. These results indicate that TFF3 is able to induce ciliogenesis and to promote airway epithelial ciliated cell differentiation, in part through an EGF-R-dependent pathway.

Distinct epithelial gene expression phenotypes in childhood respiratory allergy

“…Mucus hypersecretion and goblet cell metaplasia are important traits of asthma, also linked with IL-13 dysregulation. We also noticed that TFF3, which is expressed in mucous cells and can promote the differentiation of ciliated human airway epithelial cells [21], was up-regulated in allergic patients. This illustrates the complexity of the mechanisms leading to abnormal repair and remodelling.…”

“…This group puts together important markers of multiciliogenesis (PLK4 and CDC20B) [12] and molecules associated with asthma in genomewide analyses (GSDML and CYFIP2). A second group, which includes POSTN, SPINK5, CD44, TFF3, ITLN1, FOXA3 and SPDEF, was clearly associated with basal or mucous cells [21]. Most of them were decreased during in vitro differentiation while all were up-regulated in patients with uncontrolled asthma.…”

Molecular Mechanisms of Metaplasia, Differentiation and Hyperplasia of Goblet Cellin Allergic Asthma

“…Using a murine asthma model it was shown that trans-differentiating Clara cells specifically express TFF1, which is stored in a specific subset of secretory granules [47]. These points to a role for TFF1 as an autocrine factor for the transdifferentiation of Clara cells toward goblet cells [47]. It is demonstrated that induction of TFF3 synthesis was shown with differentiation in vivo humanized tracheal xenograft and in vitro air-liquid interface culture models.…”

“…Goblet cells are derived, at least in part, from Clara cells, which trans-differentiate from a serous into a mucous phenotype. Using a murine asthma model it was shown that trans-differentiating Clara cells specifically express TFF1, which is stored in a specific subset of secretory granules [47]. These points to a role for TFF1 as an autocrine factor for the transdifferentiation of Clara cells toward goblet cells [47].…”

Regulation of membrane-type 1 matrix metalloproteinase expression by zonula occludens-2 in human lung cancer cells

“…Human bronchial epithelial cells (HBECs) were isolated from human normal bronchial tissues from patients undergoing surgery Clin Exp Metastasis for bronchial carcinoma and were cultivated as described previously [19]. Human lung cell lines Calu-3, 16HBE, BEAS-2B, and BZR were grown in Dulbecco's modified Eagle's medium (DMEM) (Gibco, Invitrogen, Carlsbad, CA, USA) supplemented with 10 % fetal calf serum (Gibco), and 100 UI/mL of penicillin and streptomycin (Gibco), and HBE4-E6/E7 (HBE4) in keratinocyte serum free medium (Gibco) supplemented with 0.2 ng/mL EGF (epidermal growth factor) and 25 lg/mL BPE (Bovine pituitary extract).…”