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Biosynthesis and physiological roles of extremely long chain lipids in the eye

极长链脂质在眼中的生物合成和生理作用

基本信息

批准号：
9319318
负责人：
Igor A Butovich
金额：
$ 40.5万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9319318
关键词：
Affect Anabolism Animal Model Animals Ankle Antisense RNA Attention Biochemical Biochemical Pathway Biochemistry Blood Cells Characteristics Chemicals Cholesterol Confocal Microscopy Consensus Country Data Desiccation Detection Diet Dietary Fats Disease Environmental Risk Factor Enzymatic Biochemistry Enzymes Equilibrium Evaluation Eye Eye diseases Eyelid structure Fatty Acids Fatty Alcohols Film Gas Chromatography Gene Chips Gene Expression Microarray Analysis Gene Expression Profiling Gene Mutation Gene Silencing General Population Generations Genes Genetic Genomic approach Goals Health Human Impairment In Situ In Situ Hybridization Individual Knockout Mice Knowledge Label Laboratories Lead Length Lipids Liquid substance Maps Mass Spectrum Analysis Metabolic Pathway Metabolism Methods Molecular Profiling Morphology Mus Mutant Strains Mice Mutation Nature Ocular Pathology Oleic Acids Outcome Pathology Pathway interactions Pattern Physiological Physiology Polarization Microscopy Population Procedures Production Proteins Proteomics RNA Reporting Research Role Sampling Small Interfering RNA Specimen Stable Isotope Labeling Staining method Stains Structure Study models Symptoms Tarsal plate Testing Therapeutic Intervention Tissues Tracer Transgenes Validation Wild Type Mouse Work effective therapy experimental study eye dryness fatty acid biosynthesis human subject hydroxy fatty acid interest knockout gene lipid biosynthesis lipid metabolism meibomian gland meibomian gland dysfunction microcalorimetry mouse model novel strategies novel therapeutics ocular surface pi bond public health relevance reflectance confocal microscopy research clinical testing response targeted treatment technique development treatment strategy uptake

项目摘要

DESCRIPTION (provided by applicant): The fundamental goal of this project is to investigate metabolic pathways that lead to generation of lipid-rich secretion (called meibum) by Meibomian glands (MG). MG are embedded into the tarsal plates of humans and animals. Meibum protects the ocular surface from hazardous environmental factors and desiccation through its participation in formation of the tear film (TF). Abnormal quality, and/or insufficient quantity of meibum destabilize the human TF, which is one of the underlying factors of dry eye disease (DE), particularly of one of its major forms - Meibomian gland dysfunction. Numerous reports indicated that DE affects up to 30% of the general population of the USA and other countries. As a practical outcome of our research, we expect to discover new, more effective treatments for DE than those that are available now. Currently, no detailed information on the biosynthetic pathways that lead to formation of meibum is available. Our plan is to fill in this important gap i knowledge using mice as animal models. Earlier, we have discovered that lipid compositions of meibum of humans and mice are very close. Also, our pilot experiments with human and mouse MG tissue specimens demonstrated that all tested lipid-synthesizing enzymes are expressed in both species. Thus, mice are convincing (and convenient) models for studies that are ultimately targeting human ocular pathologies related to MG. As Aim 1, we propose to investigate biosynthetic pathways that lead to formation of these specific lipids, and determine which enzymatic steps and lipid products are critical for making healthy meibum. We will determine expression patterns and cellular and subcellular localization of major enzymes responsible for producing meibum, in mouse MG and adnexa. Our Aim 2 is to elucidate changes in meibum, MG, and adnexa in response to suppression or activation of specific genes related to lipid metabolism. To test our ideas, we will make use of various currently available gene knockout and transgene mouse models with known lipid abnormalities, whose ocular features have not been characterized yet because those mutant mice were developed for other purposes. Also, we will develop new approaches for temporary silencing the genes of interest using anti-sense RNA. As Aim 3, we will determine whether dietary lipids can have a major impact on meibum by tracing the uptake and metabolism of 2H- and 13C-labeled dietary lipid tracers by MG. Our goals will be achieved using a combination of chemical, biochemical, physiological and genomic approaches, which include, but are not limited to, liquid and gas chromatography, mass spectrometry, microcalorimetry, hot stage polarized light microscopy, in vivo confocal microscopy, proteomics, in-situ hybridization, immuno(histo)chemical staining, gene expression analysis, in vivo confocal microscopy, and clinical evaluation of the ocular structures of experimental mice.