Project description:We have developed a rat model of UVB-induced ocular surface and eyelid damages mimicking ocular rosacea, a common chronic inflammatory and neurovascular ocular surface disease associated with meibomian gland dysfunction. Rat eyelids were exposed to UVB for 5 minutes daily (300mJ/cm2) for 5 days. UVB-exposed rats displayed altered meibocyte activity and lipid production, ductal epithelial hyper-keratinization, apoptosis, mitochondrial dysfunction and oxidative stress. Corneal epithelial defects, barrier disruption and abnormal innervation occurred 2 weeks after UVB exposure, subsequent to meibomian gland dysfunction. The aim of this study was to investigate the transcriptomic signature associated with meibomian gland dysfunction induced by UVB in rats. We revealed pathways related to keratinization, cell cycle and proliferation, DNA damage and repair, inflammation, immune responses, oxidative responses, and fatty acid metabolism. These regulations supported the observed functional changes of meibomian gland and were consistent with the transcriptomic signature of human meibomian gland dysfunction.
Project description:Genome-wide analysis of dihydrotestosterone (DHT) induced changes in gene expression in immortalized human meibomian gland epithelial cells. Analysis of regulation of immortalized human meibomian gland epithelial cells by dihydrotestosterone at gene expression level. The hypothesis tested in the present study was that the androgen-eye interaction in ocular surface epithelial cells like meibomian gland cells is influenced by androgens through regulation of the expression of multiple genes. Results provide important information of the differential regulation of numerous genes in response to dihydrotestosterone incubation of immortalized human meibomian gland epithelial cells. Total RNA was obtained from immortalized human meibomian gland epithelial cells treated for 72 hours with 10 nM dihydrotestosterone (n=3) or vehicle (n=3). The RNA was then used with Illumina HumanHT-12 v3 Expression BeadChips to determine the effect of DHT on gene expression in an immortalized human meibomian epithelial cell line developed in our laboratory.
Project description:Genome-wide analysis of dihydrotestosterone (DHT) induced changes in gene expression in immortalized human meibomian gland epithelial cells. Analysis of regulation of immortalized human meibomian gland epithelial cells by dihydrotestosterone at gene expression level. The hypothesis tested in the present study was that the androgen-eye interaction in ocular surface epithelial cells like meibomian gland cells is influenced by androgens through regulation of the expression of multiple genes. Results provide important information of the differential regulation of numerous genes in response to dihydrotestosterone incubation of immortalized human meibomian gland epithelial cells.
Project description:While meibomian gland dysfunction (MGD) is widely recognized as a major cause of evaporative dry eye disease, little is known about normal gland differentiation and lipid synthesis or the mechanism underlying gland atrophy and abnormal lipid secretion. In this study, we used single-cell and spatial transcriptomics to probe changes in cell composition, differentiation, and gene expression associated with two murine models of MGD: age-related gland atrophy in wild-type mice and altered meibum quality in acyl-CoA wax alcohol acyltransferase 2 (Awat2) knockout (KO) mice. We identified the stratified expression of lipogenic genes during meibocyte differentiation, which may control the progressive synthesis of meibum lipids; an age-related decrease in meibocytes; and increased immune cell infiltration. Additionally, we detected unique immune cell populations in the Awat2 KO mouse suggesting the activation of psoriasis-like, inflammatory pathways associated with the synthesis of altered meibum quality causing ductal dilation and hyperplasia of the duct. Together these findings support novel mechanism controlling gland function and dysfunction.
Project description:To dissect molecular mechanisms underlying Hsd3b6−/−meibomian gland phenotypes, we performed RNA-sequencing (RNA-seq) analysis using fluorescence-activated cell sorting (FACS)-purified meibomian gland cells. RNA-seq analysis was performed using FACS-purified Itgav(+);CD45(−) meibomian gland acinar cells from wildtype and Hsd3b6−/−mice.As revealed by gene ontology (GO) enrichment analysis, cells sorted from Hsd3b6−/−mice show increased expression of inflammation-related genes. On the other hand, downregulated genes exhibit the highest enrichment score for the GO term related to “regulation of anatomical structure morphogenesis”, a signature accounting for the phenotype of the meibomian gland atrophy of Hsd3b6−/− mice.
Project description:The objective of this study is to identify human meibomian gland genes that may promote the development and/or progression of meibomian gland dysfunction. Total RNA was isolated from normals (n=6) and patients with meibomian disease (n=6). The RNA was then used in conjuction with HumanHT-12 v3 Expression BeadChips to assay differential gene expression between normal and diseased meibomian glands.
Project description:Eda signaling plays critical role for Meibomian gland dvelopment, however, the crosstalk of Eda with other signaling is largly unknown. By comparing expreession profilings between wild-type and Eda mutant Tabby eyelids, we identified Dkk4 and Lrp6 highly expressed during mouse Meibomian gland development.
Project description:The objective of this study is to identify human meibomian gland genes that may promote the development and/or progression of meibomian gland dysfunction.