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2016 Agenda and Abstracts | < Previous | Next > 2016 OMIG Abstract 15 Insight Into the Corneal Wound Healing Response: Transcriptomic and Metabolomic Analysis of Corneal Epithelial Cells Challenged by Bacteria Purpose: We recently demonstrated corneal wound healing inhibition by ocular bacterial pathogens. With the contact lens-associated keratitis pathogen Serratia marcescens, LPS was responsible for this inhibition. This study sought to determine the impact of bacterian the host response of corneal epithelial cells in vitro. Methods: Growth medium (mock) and sterile bacterial culture filtrates (secretomes) were added to human corneal limbal epithelial (HCLE) cells. HCLE supernatants from mock and secretome treated cells were collected 24 hours post challenge and analyzed by cytokine array, and ELISA. Contents of challenged HCLE cells were also metabolomically analyzed. RNA from HCLE cells with and without secretomes was analyzed by microarray and qRT-PCR. Results: HCLEs treated with S. marcescens secretomes resulted in increased IL-1α, IL-1β, GM-CSF, and IL-6 as determined by cytokine array, ELISA for IL-1 β, and microarray. Metabolomics from secretome treated HCLEs had elevated levels of phosphoethanolamine, a phosphorylated amino alcohol associated with autophagy, lipid signaling, and apoptosis. Conclusions: Using a combination of transcriptomics and metabolomics this study provides insight into the impact of bacteria on human corneal epithelial cells. Together these data support when HCLEs are treated with S. marcescens secretomes, there is an increase in pro-inflammatory markers and small molecules. The potential signaling pathways involved in microbial-induced corneal wound healing inhibition and associated biomarkers are discussed. S=Research to Prevent Blindness, Eye and Ear Institute of Pittsburgh, NIH grant EY024785, NIH grant AI085570, NIH Core Grant EY08098 2016
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