title: Global Metabonomic and Proteomic Analysis of Human Conjunctival Epithelial Cells (IOBA-NHC) in Response to Hyperosmotic Stress.
abstract: "Dry eye" is a multifactorial inflammatory disease affecting the ocular surface. Tear hyperosmolarity in dry eye contributes to inflammation and cell damage. Recent research efforts on dry eye have been directed toward biomarker discovery for diagnosis, response to treatment, and disease mechanisms. This study employed a spontaneously immortalized normal human conjunctival cell line, IOBA-NHC, as a model to investigate hyperosmotic stress-induced changes of metabolites and proteins. Global and targeted metabonomic analyses as well as proteomic analysis were performed on IOBA-NHC cells incubated in serum-free media at 280 (control), 380, and 480 mOsm for 24 h. Twenty-one metabolites and seventy-six iTRAQ-identified proteins showed significant changes under at least one hyperosmotic stress treatment as compared with controls. SWATH-based proteomic analysis further confirmed the involvement of inflammatory pathways such as prostaglandin 2 synthesis in IOBA-NHC cells under hyperosmotic stress. This study is the first to identify glycerophosphocholine synthesis and O-linked β-N-acetylglucosamine glycosylation as key activated pathways in ocular surface cells under hyperosmotic stress. These findings extend the current knowledge in metabolite markers of dry eye and provide potential therapeutic targets for its treatment.
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.|Department of Pharmacy, Faculty of Science, National University of Singapore , 18 Science Drive 4, Singapore 117543, Singapore.
authors.forename: Liyan
authors.initials: L
authors.lastname: Chen
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.|Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore , 1E Kent Ridge Road, NUHS Tower Block Level 7, Singapore 119228, Singapore.|Department of Ophthalmology, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine , 1665 Kongjiang Road, Shanghai 200092, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Institute of Molecular Systems Biology, Department of Biology, ETH Zurich , Auguste-Piccard-Hof 1, 8093 Zürich, Switzerland.
authors.forename: Tiannan
authors.initials: T
authors.lastname: Guo
authors.affiliation: Cardiovascular and Metabolic Disorders Program & Centre for Computational Biology, Duke-NUS Graduate Medical School , 8 College Road, Singapore 169857, Singapore.
authors.forename: Sujoy
authors.initials: S
authors.lastname: Ghosh
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.
authors.forename: Siew Kwan
authors.initials: SK
authors.lastname: Koh
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.|Department of Statistics and Applied Probability, Faculty of Science, National University of Singapore , 6 Science Drive 2, Singapore 117546, Singapore.
authors.forename: Dechao
authors.initials: D
authors.lastname: Tian
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.|Department of Statistics and Applied Probability, Faculty of Science, National University of Singapore , 6 Science Drive 2, Singapore 117546, Singapore.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.
authors.forename: Deyong
authors.initials: D
authors.lastname: Jia
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.|Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore , 1E Kent Ridge Road, NUHS Tower Block Level 7, Singapore 119228, Singapore.|Neuroscience and Behavioral Disorders Program, Duke-NUS Graduate Medical School , 8 College Road, Singapore 169857, Singapore.
authors.forename: Roger W
authors.initials: RW
authors.lastname: Beuerman
authors.affiliation: Institute of Molecular Systems Biology, Department of Biology, ETH Zurich , Auguste-Piccard-Hof 1, 8093 Zürich, Switzerland.|Faculty of Science, University of Zurich , Winterthurerstrasse 190, 8057 Zürich, Switzerland.
authors.forename: Ruedi
authors.initials: R
authors.lastname: Aebersold
authors.affiliation: Department of Pharmacy, Faculty of Science, National University of Singapore , 18 Science Drive 4, Singapore 117543, Singapore.
authors.forename: Eric Chun Yong
authors.initials: EC
authors.lastname: Chan
authors.affiliation: Singapore Eye Research Institute , The Academia, 20 College Road, Discovery Tower Level 6, Singapore 169856, Singapore.|Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore , 1E Kent Ridge Road, NUHS Tower Block Level 7, Singapore 119228, Singapore.|Neuroscience and Behavioral Disorders Program, Duke-NUS Graduate Medical School , 8 College Road, Singapore 169857, Singapore.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
mesh_terms: D002460:Cell Line; D003228:Conjunctiva / Q000166:cytology*; D015352:Dry Eye Syndromes; D004847:Epithelial Cells / Q000502:physiology*; D006801:Humans; D007553:Isotope Labeling; D055442:Metabolome / Q000187:drug effects / Q000502:physiology*; D055432:Metabolomics; D009997:Osmotic Pressure / Q000187:drug effects / Q000502:physiology*; D020011:Protective Agents / Q000494:pharmacology; D020543:Proteome / Q000032:analysis* / Q000187:drug effects; D040901:Proteomics
chemical_list: D020011:Protective Agents; D020543:Proteome
keywords: SWATH; dry eye; hyperosmolarity; iTRAQ; metabolomics; metabonomics; osmoprotectants; osmostic stress; proteomics