Biology subjects

Wasson, C.

Publications and source records attributed to Wasson, C..

3 recordsLinked to original sources

bioRxiv2026

The lncRNA HOTAIR/EZH2 interaction inhibitor AC1Q3QWB (AQB) attenuates fibrotic SSc skin tissue re-modelling

ObjectivesThe long non-coding RNA HOTAIR has previously been shown to drive pro-fibrotic gene expression in SSc dermal fibroblasts through its ability to interact with EZH2. Targeting EZH2 enzymatic activity can reverse HOTAIR mediated pro-fibrotic gene expression but its many functions make it an undesirable therapeutic target for SSc. Recently inhibitors selectively targeting the HOTAIR/EZH2 interaction have been developed. The aim of this study was to characterise the ability of one of these inhibitors to modulate SSc tissue remodelling. MethodsExplanted healthy and SSc dermal fibroblasts were treated with the HOTAIR/EZH2 inhibitor AC1Q3QWB (AQB) (20{micro}M) for 48 hours. In addition, healthy dermal fibroblasts were transduced with a lentivirus encoding HOTAIR or a scrambled control. Conditioned media from healthy, SSc and HOTAIR-expressing dermal fibroblasts was used to stimulate human keratinocytes (HaCaTs). Scramble control and HOTAIR expressing fibroblasts were grown in 3D skin equivalents containing primary keratinocytes and keratin 9 (K9) immunohistochemistry performed. ResultsAQB inhibits pro-fibrotic gene expression in HOTAIR expressing dermal fibroblasts, validating the specificity of the inhibitor. In SSc patient dermal fibroblasts, AQB blocked pro-fibrotic gene expression but did not affect gene expression in healthy dermal fibroblasts. SSc patient skin was shown to express high levels of the palmoplantar specific K9 and Epithelial to Mesenchymal transition (EMT) markers. Through co-culture experiments we showed these effects were mediated by SSc dermal fibroblasts. This tissue remodelling was disrupted when HOTAIR/EZH2 interaction was inhibited in the fibroblasts with AQB. ConclusionsWe have shown for the first time that directly inhibiting HOTAIR/EZH2 interaction blocks pro-fibrotic gene expression in SSc fibroblasts and tissue re-modelling found in SSc patient skin. This may represent a novel therapeutic intervention.

Cell Biology↗

bioRxiv2020

Targeting BDCA2 suppresses Plasmacytoid dendritic cell dependent interferon response and tissue fibrosis

Plasmacytoid dendritic cells (pDC) have been implicated in the pathogenesis of Scleroderma (SSc) through their ability to infiltrate the skin and secrete interferons (IFN) and proinflammatory chemokines. Blood Dendritic Cells Antigen 2 (BDCA2) is an inhibitory type II C-type lectin expressed by human pDC. Here we determined the effects of BDCA2 internalisation on pDC mediated skin inflammation and fibrosis in human preclinical models of skin inflammation and fibrosis in vitro and in vivo. BDCA2 targeting reversed TLR-signalling induced transcriptome and differentiation of pDC and suppressed their ability to induce IFN response in organotypic 3D human skin cultures in vitro. In vivo, xenotransplantation of human pDC into immunocompromised mice (XenoSCID) significantly increased IFN induced responses to topical TLR7/9 agonist and separately enhanced the fibrotic response to bleomycin. Targeting of BDCA2 strongly suppressed both of these pathological responses ameliorating skin inflammation and fibrosis. Together, these preclinical data strongly support the notion that human pDC play a key role in immune driven skin fibrosis, which can be effectively blocked by targeting BDCA2.

immunology↗

bioRxiv2018

High-risk human papillomaviruses down-regulate expression of the Ste20 family kinase MST1 to inhibit the Hippo pathway and promote transformation

Human papillomaviruses (HPV) are a major cause of malignancy worldwide They are the aetiological agent of almost all cervical cancers and an increasing number of head and neck carcinomas. Deregulation of the Hippo pathway component YAP1 has recently been demonstrated to play a role in HPV-mediated cervical cancer, but whether other components of this pathway are implicated in the pathogenesis of this disease remains poorly understood.\n\nThe expression level and activation status of critical Hippo pathway components were analysed across multiple cytology samples from patients with cervical disease, as well as HPV positive (HPV+) and HPV negative (HPV-) cervical cancer cell lines using real time qPCR, western blot and immunohistochemistry. In parallel, we assessed the effects of MST1 and MST2 overexpression upon cervical cancer cell proliferation, migration and invasion. Finally, we interrogated the consequences of interrupted MST1 and MST2 function using a targeted small molecule inhibitor in tandem with kinase inactive MST mutants. Our analysis found that expression of the Ste20 kinase MST1 was decreased within both HPV+ primary patient samples and cervical cancer cell lines. This effect was mediated by the virus-coded oncoproteins E6 and E7, which impair MST1 transcription. Reintroduction of MST1, or its paralogue MST2, into HPV positive cervical cancer cells re-activated the Hippo pathway, leading to a reduction in cell proliferation, migration and invasion. Finally, using a small molecule inhibitor of MST1/2 or kinase inactive mutants of either protein, we demonstrated that this effect required the kinase function of MST1/2. Our results reveal that HPV down regulates MST1 expression to inactivate the Hippo pathway and so drive cells towards transformation.

microbiology↗