Project description:<p>Transcription factor p63 is a key regulator of epidermal keratinocyte proliferation and differentiation. Mutations in the p63 DNA-binding domain are associated with Ectrodactyly Ectodermal Dysplasia Cleft Lip/Palate (EEC) syndrome. Underlying molecular mechanism of these mutations however remain unclear. Here we characterized the transcriptome and epigenome of p63 mutant keratinocytes derived from EEC patients. The transcriptome of p63 mutant keratinocytes deviated from the normal epidermal cell identity. Epigenomic analyses showed an altered enhancer landscape in p63 mutant keratinocytes contributed by loss of p63-bound active enhancers and by unexpected gain of enhancers. The gained enhancers were frequently bound by deregulated transcription factors such as RUNX1. Reversing RUNX1 overexpression partially rescued deregulated gene expression and the altered enhancer landscape. Our findings identify an unreported disease mechanism whereby mutant p63 rewires the enhancer landscape and affects epidermal cell identity, consolidating the pivotal role of p63 in controlling the enhancer landscape of epidermal keratinocytes.</p>
Project description:To identify genes regulated by BAF53A, we carried out RNA-seq analysis in three SCC cell lines after BAF53A knockdown by siRNA and in normal keratinocytes with BAF53A overexpression.
Project description:Hsa_circ_0084443 expression level is down-regulated during normal skin wound healing and higher level of hsa_circ_0084443 was found in chronic non-healing diabetic foot ulcers compared to normal wounds. However, the biological function of hsa_circ_0084443 in epidermal keratinocytes during wound repair has not been studied. To study the genes regulated by hsa_circ_0084443, we transfected siRNA targeting hsa_circ_0084443 diagnostic junction into human primary epidermal keratinocytes to knockdown hsa_circ_0084443 expression. We performed a global transcriptome analysis of keratinocytes upon knockdown of hsa_circ_0084443 using Affymetrix arrays.
Project description:The study aimed to interrogate whether the 1,4-naphtoquinone derivative lawsone activates the aryl hydrocarbon receptor (AhR) downstream transcriptional program in keratinocytes and is capable of modulating skin homeostasis. The study has been performed on human primary epidermal keratinocytes (HEKs), which represent the first line of defense against exogenous molecules. HEK cells were treated with the vehicle control DMSO (control), stimulated with lawsone or with the TLR2 ligand Pam2CSK4. Transcriptional profiles of HEK cells were studied to investigate the regulation of AhR -related genes, Nrf2-related genes and epidermal differentiation and keratin genes. We demonstrated that lawsone was sensed by keratinocytes and activated AhR. In particular, lawsone efficiently activated the transcriptional program of AhR and promoted keratinocyte differentiation.
Project description:Three types of oral cancer cell lines (HSC3,Sa3 and SAS) and human normal oral keratinocytes(HNOKs) were used to identify a circular RNA specifically. expressed in oral cancer
Project description:To identify accessible chromatin regions regulated BAF53A in squamous cell carcinomas (SCCs), we performed ATAC-seq in FaDu SCC cell lines after BAF53A knockdown by siRNA and in normal keratinocytes with BAF53A overexpression.
Project description:The morphology and the behavior of skin and oral tissue keratinocytes are different. One significant dissimilarity between the two sites is the response to injury. Oral and skin keratinocytes have intrinsic differences in the response to injury and such differences are reflected in gene expression profiles. We used microarrays to investigate differences in global gene expression patterns between baseline skin and oral epithelium sheets without their underlying connective tissue. Paired skin and oral epithelium was separated from the dermis for RNA extraction and hybridization on Affymetrix microarrays. Skin epidermal tissues were obtained from the tail of mice and oral epidermal tissues were obtained from the hard palate. Enzymatically isolated epithelium was used for analysis.