Hong Wan

Queen Mary, University of London | QMUL

The role of desmoglein-3 (DSG3) in oncogenesis is unclear. This study aimed to uncover molecular mechanisms through comparative transcriptome analysis in oral cancer cells, defining potential key genes and associated biological processes related to DSG3 expression. Four mRNA libraries of oral squamous carcinoma H413 cell lines were sequenced, and 5...

The role of desmoglein-3 (DSG3) in oncogenesis is unclear. This study aimed to uncover molecular mechanisms through comparative transcriptome analysis in oral cancer cells, defining potential key genes and associated biological processes related to DSG3 expression. Four mRNA libraries of oral squamous carcinoma H413 cell lines were sequenced and 59...

The Hippo pathway is an evolutionarily conserved pathway that serves to promote cell death and differentiation while inhibiting cellular proliferation across species. The downstream effectors of this pathway, yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ), are considered vital in promoting the output of t...

Simple Summary The early treatment of oral cancer is a high priority, as improvements in this area could lead to greater cure rates and reduced disability due to extensive surgery. Oral cancer is very difficult to detect in over 70% of cases as it develops unseen until quite advanced, sometimes rapidly. It has become apparent that there are at leas...

The Hippo-YAP pathway serves as a central signalling hub in epithelial tissue generation and homeostasis. Yes-associated protein (YAP) is an essential downstream transcription cofactor of this pathway, with its activity being negatively regulated by Hippo kinase-mediated phosphorylation, leading to its cytoplasmic translocation or degradation. Our...

Potentially pre-malignant lesions (PPOLs) are composed of keratinocytes that are either mortal (MPPOL) or immortal (IPPOL) in vitro . We report here that MPPOL, but not generally IPPOL keratinocytes upregulate various extracellular tumor promoting cytokines (interleukins 6 and 8) and prostaglandins E1 (ePGE1) and E2 (ePGE2) relative to normal oral...

Alterations of the Hippo-YAP pathway are potential targets for oral squamous cell carcinoma (OSCC) therapy, but heterogeneity in this pathway could be responsible for therapeutic resistance. We analysed the Hippo-YAP signatures in a cohort of characterised keratinocyte cell lines derived from the mouth floor and buccal mucosa from different stages...

The immunobullous condition Pemphigus Vulgaris (PV) is caused by autoantibodies targeting the adhesion proteins of desmosomes, leading to blistering in the skin and mucosal membrane. There is still no cure to the disease apart from the use of corticosteroids and immunosuppressive agents. Despite numerous investigations, the pathological mechanisms...

Pemphigus Vulgaris (PV) is a life-threatening autoimmune disease manifested with blisters in the skin and mucosa and caused by autoantibodies against adhesion protein desmoglein-3 (Dsg3) expressed in epithelial membrane linings of these tissues. Despite many studies, the pathogenesis of PV remains incompletely understood. Recently we have shown Dsg...

The Yes-associated protein (YAP) is a downstream effector of the Hippo pathway and acts as a key transcription co-factor to regulate cell migration, proliferation, and survival. The Hippo pathway is evolutionarily conserved and controls tissue growth and organ size. Dysregulation and heterogeneity of this pathway are found in cancers, including ora...

Background: Behçet's disease is a rare, multisystem vasculitis disease characterized by recurrent orogenital ulcerations with its etiology remained unclear. The transcription factor p53 has been reported to be upregulated in some autoimmune diseases, such as lupus erythematosus, dermatomyositis, and psoriasis. However, little is known about its al...

Desmoglein 3 (Dsg3) plays a crucial role in cell-cell adhesion and tissue integrity. Increasing evidence suggests that Dsg3 acts as a regulator of cellular mechanotransduction, but little is known about its direct role in mechanical force transmission. The present study investigated the impact of cyclic strain and substrate stiffness on Dsg3 expres...

Desmoglein 3 (Dsg3), plays a crucial role in cell-cell adhesion and tissue integrity. Increasing evidence suggests that Dsg3 acts as a regulator of cellular mechanotransduction, but little is known about its direct role in mechanical force transmission. The present study investigated the impact of cyclic strain and substrate stiffness on Dsg3 expre...

Desmoglein-3 (Dsg3), the Pemphigus Vulgaris (PV) antigen (PVA), plays an essential role in keratinocyte cell–cell adhesion and regulates various signaling pathways involved in the progression and metastasis of cancer where it is upregulated. We show here that expression of Dsg3 impacts on the expression and function of p53, a key transcription fact...

Kidney disease prevalence increases with age, with a common feature of the disease being defects in the epithelial tight junctions. Emerging evidence suggests that the desmosomal adhesion protein Desmoglein-3 (Dsg3) functions beyond the desmosomal adhesion and plays a role in regulating the fundamental pathways that govern cell fate decisions in re...

Desmoglein-3 (Dsg3), the Pemphigus Vulgaris (PV) antigen (PVA), plays an essential role in keratinocyte cell-cell adhesion and regulates various signaling pathways implicated in the pathogenesis the PV blistering disease. We show here that expression of Dsg3 may directly influence p53, a key transcription factor governing the response to cellular s...

Desmoglein 3 (Dsg3) is an adhesion receptor in desmosomes, but its role in carcinoma cell migration and invasion is mostly unknown. Our aim was to quantitatively analyse the motion of Dsg3-modified carcinoma cells in 2D settings and in 3D within tumour microenvironment mimicking (TMEM) matrices. We tested mutant constructs of C-terminally truncated...

Adherens junctions and desmosomes confer strong adhesion and thus occur with high frequency in epithelia that are subject to extensive mechanical stress. Both anchoring junctions are linked to the cytoskeletal filaments and provide scaffolds for the maintenance of tissue integrity. The adhesion cores of all junctions consist of the transmembrane pr...

Cell-cell interactions are essential in many physiological processes of epithelia and are often found to be deregulated in a variety of human diseases, including infectious, autoimmune, hereditary conditions as well as cancers. In principle, there are three functional groups of intercellular junctions, known as the tight junction, gap junction and...

Immunofluorescence (IF) is a technique widely used in medical research and diagnostics due to its simplicity, accuracy, and rapidity. In this article, we present the applications of IF in the diagnosis of seven oral diseases; namely, pemphigus vulgaris, mucous membrane pemphigoid, linear IgA disease, oral lichen planus, oral mucosal lupus erythemat...

E-cadherin mediated cell-cell adhesion plays a critical role in epithelial cell polarization and morphogenesis. Our recent studies suggest that the desmosomal cadherin, desmoglein 3 (Dsg3) cross talks with E-cadherin and regulates its adhesive function in differentiating keratinocytes. However, the underlying mechanism remains not fully elucidated....

Desmosomes are one of the specialised intercellular junctions and are essential for the maintenance of cellular and tissue integrity. Desmoglein 3 (Dsg3) is a member of the desmoglein subfamily that serves as the adhesion receptors in desmosomes. Dsg3 is predominantly expressed in stratified epithelia and recently has been implicated in cancer cell...

This data article contains extended, complementary analysis related to the research articles entitled “Desmoglein 3, via an interaction with E-cadherin, is associated with activation of Src” (Tsang et al., 2010) [1] and figures related to the review article entitled “Desmoglein 3: a help or a hindrance in cancer progression?” (Brown et al., 2014) [...

Desmoglein 3 is one of seven desmosomal cadherins that mediate cell-cell adhesion in desmosomes. Desmosomes are the intercellular junctional complexes that anchor the intermediate filaments of adjacent cells and confer strong cell adhesion thus are essential in the maintenance of tissue architecture and structural integrity. Like adherens junctions...

Desmosomes are one of the specialised intercellular junctions and are essential for the maintenance of cellular and tissue integrity. Desmoglein 3 (Dsg3) is a member of the desmoglein subfamily that serves as the adhesion receptors in desmosomes. Dsg3 is predominantly expressed in stratified epithelia and recently has been implicated in cancer cell...

Desmoglein 3 (Dsg3), the pemphigus vulgaris antigen, has recently been shown to be upregulated in squamous cell carcinoma (SCC) and has been identified as a good tumor-specific marker for clinical staging of cervical sentinel lymph nodes in head and neck SCC. However, little is known about its biological function in cancer. The actin-binding protei...

Histopathological discordance with molecular phenotype of many human cancers poses clinically challenging tasks for accurate cancer diagnosis which impacts on treatment strategy and patient outcome. Hence, an objective, accurate and quantitative method is needed. A quantitative Malignancy Index Diagnostic System (qMIDS) was developed based on 14 FO...

Desmoglein 3 (Dsg3), a member of the desmoglein sub-family, serves as an adhesion molecule in desmosomes. Our previous study showed that overexpression of human Dsg3 in several epithelial lines induces formation of membrane protrusions, a phenotype suggestive of Rho GTPase activation. Here we examined the interaction between Dsg3 and actin in detai...

Keratin 15 (K15) is a type I keratin that is used as a marker of stem cells. Its expression is restricted to the basal layer of stratified epithelia, and the bulge in hair follicles. However, in certain clinical situations including oral lichen planus, K15 is induced in suprabasal layers, which is inconsistent with the role of a stem cell marker. T...

Induction of PKCδ and PKCη transcription in suspension culture. N-Terts grown in SFM were suspended in DMEM +10% FCS containing 1.3% (w/v) methylcellulose. After 24 h the cells were harvested to determine the mRNA levels of PKCα, PKCδ, PKCη, PKCζ and PKCε by qPCR. Each bar represents the mean±SEM where n = 3. (P<0.01, very significant, **; P<0.001,...

Differentiation of primary human keratinocytes in suspension. (a) N-Terts and (b) normal primary human epidermal keratinocytes growing in RM+ +10% FCS were suspended in DMEM +20% FCS containing 1.3% (w/v) methylcellulose. At different time intervals, cells were harvested and used to determine mRNA expression by qPCR. The RM+ medium contains 10 ng/m...

Induction of K15 transcription in HaCaT at high cell density. (a) HaCaT keratinocytes were grown at low (30%) and high (95%) confluence at low calcium (0.09 mM) concentration for 24 h after which the cells were lysed to determine the mRNA expression. (b) The cells grown at low (S) and high (C) densities were analyzed by western blotting for express...

Oligonucleotides used in this study. (TIF)

E-cadherin, a classical cadherin, is an adhesion receptor in adherens junctions and has important functions in cell-cell adhesion and cell signalling. Recently we reported that a desmosomal cadherin, desmoglein 3 (Dsg3), an autoantigen in pemphigus vulgaris (PV), associates with E-cadherin and activates Src, which results in tyrosine phosphorylatio...

Objectives: Desmoglein 3 (Dsg3) is a member of desmosomal cadherins that plays a pivotal role in cell-cell adhesion and the maintenance of tissue integrity. Desmosomal cadherins have the ability of suppressing the invasiveness of cells in culture and the reduction of these proteins is frequently seen in cancer, the event that may facilitate tumour...

Objectives: Desmoglein 3 (Dsg3) is a desmosomal adhesion protein expressed in basal and immediate suprabasal layers of skin. Importance of Dsg3 in cell–cell adhesion and maintenance of tissue integrity is illustrated by findings of keratinocyte dissociation in the autoimmune disease, pemphigus vulgaris, where autoantibodies target Dsg3 on keratinoc...

Control images for the AbFRET experiments, showing the level of bleed through into the FRET channel of cells labelled with the Donor alone and Acceptor alone. The FRET channel is shown using the Spectrum look up table from Image J. (9.53 MB TIF)

Expression of Dsg3 caused faster and wider spreading of 293T cells. Cells were transfected transiently with either the pBABE-GFP or -hDsg3.myc construct for two days before harvesting with trypsin. Cells then were seeded at low density onto coverslips without coating and fixed at various time points prior to immunostaining for the indicated protein...

Over-expression of Dsg3 failed to enhance cell-cell adhesion. Cells were grown to confluence before being treated with 2.4 units/ml dispase for 20 minutes to detach the epithelial sheets. The epithelial sheets were washed with PBS, twice gently, before being subjected to mechanical stress by pipetting three-five times with 1-ml blue tips. The epith...

Desmoglein 3 (Dsg3), a desmosomal adhesion protein, is expressed in basal and immediate suprabasal layers of skin and across the entire stratified squamous epithelium of oral mucosa. However, increasing evidence suggests that the role of Dsg3 may involve more than just cell-cell adhesion. To determine possible additional roles of Dsg3 during epithe...

Stem cells are permanent residents of tissues and thought to be targets of cancer initiation. The frequent, and often early, upregulation of the FOXM1 transcription factor in the majority of human cancers suggests that it may participate in the initiation of human tumorigenesis. However, this hypothesis has not been tested. Herein, we show that tar...

J Oral Pathol Med (2011) 40: 67–76 Desmosomes, the intercellular junctions that confer strong adhesion between epithelial cells, are frequently altered in malignancy. However, a comprehensive analysis of these structures has not been carried out in oral neoplasia. Oral squamous cell carcinomas (SCCs) and pre-malignant dysplasia can be sub-classifie...

Desmoplakin is a ubiquitous component of desmosomes and desmosome-like structures, such as the cardiomyocyte area composita. Two major isoforms, desmoplakin I (DSPI) and desmoplakin II (DSPII) are encoded by alternative mRNA transcripts differentially spliced from the same gene. The resulting proteins are identical in amino acid sequence with the e...

Upregulation of FOXM1 protein in oral premalignant and HNSCC cell lines. Immunofluorescence imaging of FOXM1 (FITC, green) and DNA (DAPI, blue) in normal (NHOK1, HOK365), dysplasia (D20, POE9n) and HNSCC lines (5PT, UK1). (3.37 MB TIF)

Upregulation of FOXM1B protected SVpgC2a cells from nicotine-induced cell death. (A) Fluorescence microscopy showing green fluorescence expression of EGPF or EGFP-FOXM1B expressing SVpgC2a, SqCC/Y1 and SCC25 cells, either treated with vehicle control (H20) or nicotine (10 mM) for 24 hours prior to experiment. Corresponding brightfield images show r...

FOXM1B overexpression alone in the premalignant oral keratinocyte SVpgC2a cells induces LOH in chromosome 13. Magnified view of LOH profiles in chromosome 13 across the 8 SVFN transformed clones with a list of randomly selected genes found within the consensus LOH loci in this chromosome. (8.00 MB TIF)

ChIP-qPCR data for promoters of cyclinB1/CCNB1, VEGF and p16/CDKN2A. Quantitative PCR data showing the relative copy number of promoters of CCNB1 (A), VEGF (B–C, promoter region 1 and 2) and CDKN2A (D) in the three ChIP fractions (anti-GAPDH, anti-cMYC or anti-FOXM1) on mock, EGFP or FOXM1B-transduced oral keratinocytes. *P<0.05, **P<0.01 and ***P<...

Bioinformatics analysis of CEP55 gene expression level from a published microarray dataset (GEO accession: GDS1477) comparing BT-20 breast cancer cells expressing either mock, siGFP or siFOX at 48 h post-transfection (Wonsey and Follettie, 2005). (0.61 MB TIF)

ChIP-qPCR for promoters of CEP55 and HELLS. Representative qPCR fluorescence curves and melting analysis of the promoters of CEP55 (A) and HELLS (B) on ChIP (immunoprecipitate/IP; with either anti-GAPDH, anti-cMYC or anti-FOXM1) and input (non-IP) fractions on mock, EGFP or FOXM1B-transduced oral keratinocytes. (0.54 MB TIF)

Primer sequences for ChIP-qPCR. qPCR primers were designed within 1 kilobase upstream from +1 of each gene. For CEP55 and HELLS promoters, primers were designed to encompass putative FOXM1 binding site located using ClustalW2 sequence alignment tool (see Methods). (0.25 MB TIF)

FOXM1-isoform specific real-time quantitative RT-PCR (qPCR). (A) A schematic diagram showing the relative location of primers specific to each FOXM1 isoform. (B) Primer sequences. (C) qPCR amplification curves demonstrating the specificity of each isoform-specific primer pairs in the presence of different FOXM1 isoform DNA templates (106 copies wer...

Bioinformatics analysis of FOXM1, CEP55 and HELLS gene expression level from a published microarray dataset (GEO accession: GDS1548). Analysis of oral squamous cell carcinoma (OSCC) cells from 16 patients and 4 healthy normal oral tissue samples. OSCC cells were isolated from tumours by laser capture microdissection. Results identify a strong corre...

Cancer associated with smoking and drinking remains a serious health problem worldwide. The survival of patients is very poor due to the lack of effective early biomarkers. FOXM1 overexpression is linked to the majority of human cancers but its mechanism remains unclear in head and neck squamous cell carcinoma (HNSCC). FOXM1 mRNA and protein expres...

IntroductionWhat Happens During Sensitization to Inhaled Allergens?Structure of the Airway Mucosa and its Relevance to Allergic SensitizationHypothesis of Intercellular Junctions as Targets for House-Dust Mite AllergensMolecular and Functional Biological Properties of House-Dust Mite AllergensHouse-Dust Mite Proteinase Allergens and Epithelial Morp...

The intercellular adhesive junction desmosomes are essential for the maintenance of tissue structure and integrity in skin. Desmoplakin (Dp) is a major obligate plaque protein which plays a fundamental role in anchoring intermediate filaments to desmosomal cadherins. Evidence from hereditary human disease caused by mutations in the gene encoding Dp...

We showed previously that primary keratinocytes selected for low desmoglein 3 (Dsg3) expression levels exhibited increased colony-forming efficiency and heightened proliferative potential relative to cells with higher Dsg3 expression levels, characteristics consistent with a more “stem/progenitor cell-like” phenotype. Here, we have confirmed that D...

Several hereditary human diseases are now known to be caused by distinct mutations in genes encoding various desmosome components. Although the effects of some of these mutant genes have been analysed by targeted disruption experiments in mouse models, little is known about the cell and tissue changes in affected human patients. To investigate the...

No single method has been universally adopted for identifying and isolating epidermal stem/progenitor cells, and the emergence of new markers of stem cell populations is worth exploring. Here we report, for the first time, that clusters of basal keratinocytes at the tips of the rete ridges in human palm, previously recognised as a major repository...

Hereditary skin disorders resulting from desmosome gene pathology may preferentially involve the palms and soles. Why this is so is not clear. Moreover, even in normal control skin it is unknown whether there are differences in desmosome number, size or structural organization in palmoplantar sites compared with skin from other body regions. Theref...

Ablation of the desmosomal plaque component plakophilin 1 underlies the autosomal recessive genodermatosis, skin fragility-ectodermal dysplasia syndrome (OMIM 604536). Skin from affected patients is thickened with increased scale, and there is loss of adhesion between adjacent keratinocytes, which exhibit few small, poorly formed desmosomes. To inv...

The striate form of palmoplantar keratoderma is a rare autosomal dominant disorder affecting palm and sole skin. Genetic heterogeneity of striate palmoplantar keratoderma has been demonstrated with pathogenic mutations in the desmosomal proteins desmoplakin and desmoglein 1. We have studied a four-generation family of British descent with striate p...

Lipoid proteinosis (LP), also known as hyalinosis cutis et mucosae or Urbach-Wiethe disease (OMIM 247100) is a rare, autosomal recessive disorder typified by generalized thickening of skin, mucosae and certain viscera. Classical features include beaded eyelid papules and laryngeal infiltration leading to hoarseness. Histologically, there is widespr...

Lipoid proteinosis (LP), also known as hyalinosis cutis et mucosae or Urbach-Wiethe disease (OMIM 247100) is a rare, autosomal recessive disorder typified by generalized thickening of skin, mucosae and certain viscera. Classical features include beaded eyelid papules and laryngeal infiltration leading to hoarseness. Histologically, there is widespr...

The constitutive desmosomal plaque protein desmoplakin plays a vital part in keratinocyte adhesion in linking the transmembranous desmosomal cadherins to the cytoplasmic keratin filament network. Recently, mutations in desmoplakin have been shown to underlie some cases of the autosomal dominant disorder, striate palmoplantar keratoderma, as well as...

There have been only a few studies of how allergens cross the airway epithelium to cause allergic sensitization. House dust mite fecal pellets (HDMFP) contain several proteolytic enzymes. Group 1 allergens are cysteine peptidases, whilst those of groups 3, 6 and 9 have catalytic sites indicative of enzymes that mechanistically behave as serine pept...

Fel d 1, an important allergen from domestic cats, is a significant cause of asthma. In addition to directly promoting IgE synthesis, other biological activities of allergens may contribute to either allergic sensitization or the magnitude of allergic effector responses. For example, allergens that degrade proteins have been suggested to facilitate...

Tight junctions (TJs) make a vital contribution to the barrier properties of the airway lining. Opening of TJs, or their frank cleavage, is suspected as a pathophysiological event in the lung, but research into the cellular and molecular mechanisms involved has been impeded by technical limitations of available experimental models. The authors have...

House dust mite allergen Der p 1 is a cysteine peptidase. Previously, we have suggested that the proteolytic activity of this allergen may contribute to asthma by damaging the barrier formed by the airways epithelium. The present study applied novel techniques to compare changes in permeability with quantitative events in tight junctions (TJs) and...

House dust mite (HDM) allergens are important factors in the increasing prevalence of asthma. The lung epithelium forms a barrier that allergens must cross before they can cause sensitization. However, the mechanisms involved are unknown. Here we show that the cysteine proteinase allergen Der p 1 from fecal pellets of the HDM Dermatophagoides ptero...

Asthma is a debilitating and life-threatening disease which is now the most common chronic disease of childhood. Sensitization of the lung to airbone allergens such as faecal pellet proteins of the house dust mite (HDM) is a primary risk factor for the development of asthma [1–5]. Allergic sensitization involves detection of an allergen by antigen-...

Allergenic and non-allergenic proteinases from house dust mites (HDM) cause loss of adhesion between airway epithelial cells that may result in a loss of functional cohesion between the cells and thus assist in allergen presentation. Improved cellular assay systems are needed to ascertain the mechanisms involved. To survey a series of epithelial ce...

House dust mite (HDM) allergens with cysteine and serine proteinase activity are risk factors for allergic sensitization and asthma. A simple method to fractionate proteinase activity from HDM faecal pellets into cysteine and serine class activity is described. Both proteinase fractions increased the permeability of epithelial cell monolayers. The...

The two photon microscope provides optical sectioning of fluorescent specimens with a resolution comparable to that obtained in confocal microscopy (see refs 2,3). However, the excited volume in 2-photon microscopy is limited to the focal volume (unlike conventional fluorescence microscopy where excitation occurs throughout the specimen). This mean...

Objectives: Desmoglein 3 (Dsg3) is a member of desmoglein subfamily and contributes to cell-cell adhesion in desmosomes. It was first characterized as an autoantigen in pemphigus vulgaris, an autoimmune blistering disease preferentially manifested in oral mucosa. Our recent study suggests that Dsg3 has a role beyond desmosome adhesion, being asso...

Thesis (Ph. D.)--University of Liverpool, 1995.