نبذة مختصرة : The 62nd Annual Medical Student Research Forum at UT Southwestern Medical Center (Tuesday, January 30, 2024, 3-6 p.m., D1.700 Lecture Hall) ; Each year the Medical Student Research Program awards students for the best oral presentation and the best poster presentation as judged by faculty across campus. This author received an award as one of the best poster presentations at this forum. ; INTRODUCTION: Pseudomonas aeruginosa (PA) keratitis is a severe infection of the cornea that can lead to blindness. Studies in our lab have shown that PA exploits autophagy, a major cellular degradation process, in corneal epithelial cells (hTCEpi cells) to promote intracellular survival. We have further shown that the inhibition of autophagy by the Unc 51-like kinase (Ulk1), an enzyme that mediates formation of the autophagosome, reduces intracellular levels of PA. More recently, we have demonstrated that PA infection negatively impacts host mitochondria. ULK1/2 has been reported to translocate to mitochondria to mediate mitophagy however, a role for ULK1/2 in mitochondrial homeostasis during infection has not yet been explored. In this study, we investigated the effects of the inhibition of Ulk1 during PA infection on host mitochondria. METHODS: Telomerase-immortalized human corneal epithelial (hTCEpi) cells were used for this study. Cells were cultured in serum-free defined keratinocyte media with growth supplements. Cells were inoculated with 106 CFU/ml of PA in log growth phase with or without treatment with 1 ï�M of the Ulk1/2 inhibitor MRT68921. Intracellular levels of PA were quantified using a gentamicin survival assay. Oxygen consumption and mitochondrial polarization were assessed using Seahorse metabolic flux analysis and tetraethyl-benzimidazolyl-carbocyanine iodide (JC-1), respectively. Levels of pro-inflammatory cytokines were assessed using ELISA. Untargeted metabolomics was performed using mass spectrometry. Cellular changes were further evaluated using transmission electron microscopy (TEM). RESULTS: PA ...
Relation: 62nd Annual Medical Student Research Forum; Abdallah, J., Ayilam Ramachandran, R., & Robertson, D. M. (2024, January 30). Effect of ULK1 inhibition on corneal epithelial cells during Pseudomonas aeruginosa infection [Poster session]. 62nd Annual Medical Student Research Forum, Dallas, Texas. https://hdl.handle.net/2152.5/10263; https://hdl.handle.net/2152.5/10263
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