Regulatory role of miR-146a in corneal epithelial wound healing via its inflammatory targets in human diabetic cornea |
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| Institution: | 1. Eye Program, Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA;2. Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA;3. Department of Medicine, University of Southern California, Los Angeles, CA, USA;4. David Geffen School of Medicine at UCLA, Los Angeles, CA, USA;1. Rothschild Foundation Hospital, Paris, France;2. Paris University, Paris, France;1. State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangdong Provincial Clinical Research Center for Ocular Diseases, Guangzhou, 510060, China;2. Department of Hematology, Nanfang Hospital, Southern Medical University, No. 1838, North Guangzhou Avenue, Guangzhou, Guangdong, 510515, China;1. Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller, School of Medicine, Miami, USA;2. Department of Ophthalmology, Miami Veterans Administration Medical Center, Miami, FL, USA;3. Department of Ophthalmology and Visual Science, Yale School of Medicine, New Haven, CT, USA;4. Ophthalmic Center, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China;1. Department of Ophthalmology, New Zealand National Eye Centre, The University of Auckland, New Zealand;2. Optometry and Vision Sciences, College of Health and Life Sciences, Aston University, Birmingham, UK;3. Academic Unit of Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Birmingham and Midland Eye Centre, Birmingham, UK;4. Thea Pharmaceuticals, UK |
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| Abstract: | PurposeMiR-146a upregulated in limbus vs. central cornea and in diabetic vs. non-diabetic limbus has emerged as an important immune and inflammatory signaling mediator in corneal epithelial wound healing. Our aim was to investigate the potential inflammation-related miR-146a target genes and their roles in normal and impaired diabetic corneal epithelial wound healing.MethodsOur previous data from RNA-seq combined with quantitative proteomics of limbal epithelial cells (LECs) transfected with miR-146a mimic vs. mimic control were analyzed. Western blot and immunostaining were used to confirm the expression of miR-146a inflammatory target proteins in LECs and organ-cultured corneas. Luminex assay was performed on conditioned media at 6- and 20-h post-wounding in miR-146a mimic/inhibitor transfected normal and diabetic cultured LECs.ResultsOverexpression of miR-146a decreased the expression of pro-inflammatory TRAF6 and IRAK1 and downstream target NF-κB after challenge with lipopolysaccharide (LPS) or wounding. Additionally, miR-146a overexpression suppressed the production of downstream inflammatory mediators including secreted cytokines IL-1α, IL-1β, IL-6 and IL-8, and chemokines CXCL1, CXCL2 and CXCL5. These cytokines and chemokines were upregulated in normal but not in diabetic LEC during wounding. Furthermore, we achieved normalized levels of altered secreted cytokines and chemokines in diabetic wounded LEC via specific inhibition of miR-146a.ConclusionOur study documented significant impact of miR-146a on the expression of inflammatory mediators at the mRNA and protein levels during acute inflammatory responses and wound healing, providing insights into the regulatory role of miR-146a in corneal epithelial homeostasis in normal and diabetic conditions. |
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| Keywords: | Chemokines Cytokines Diabetic cornea Limbal stem cells microRNA miR-146a NF-κB inflammatory pathway Wound healing |
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