The Role of MTF-1 as a Mediator of Ocular Toxicity

MTF-1 作为眼毒性介质的作用

• 批准号: 9014839
• 负责人: Matthew J. Jenny
• 金额: $ 21.94万
• 依托单位: UNIVERSITY OF ALABAMA IN TUSCALOOSA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9014839
• 关键词: Adult; Aging-Related Process; Animal Model; Animals; Basic Science; Binding; Binding Proteins; Biological Assay; Blindness; Cadmium; Cataract; Cell Line; Cells; Cellular Stress; ChIP-seq; Chromatin; Cigarette Smoker; Congenital Abnormality; DNA; DNA Binding; DNA Damage; Data; Defect; Development; Disease; Dominant-Negative Mutation; Drug Metabolic Detoxication; Embryo; Environment; Environmental Risk Factor; Epithelial Cells; Exposure to; Eye; Eye Development; Eye diseases; Fertilization; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genomics; Goals; Homeostasis; Hour; Human; In Situ Hybridization; In Vitro; Laboratories; Lead; Lens Opacities; Lens development; Leukocytes; Light; Link; Liver; Macular degeneration; Maintenance; Mediator of activation protein; Metal exposure; Metals; Modeling; Molecular; Molecular Profiling; Organogenesis; Orthologous Gene; Oxidative Stress; Pathology; Pharmacologic Substance; Phenotype; Play; Poison; Population Decreases; Public Health; Regulation; Regulatory Pathway; Research; Research Project Grants; Response Elements; Retinal; Risk; Role; Sampling; Sequence Analysis; Signal Transduction; Smoking; Staging; Steroids; Stress; Study models; Sunlight; System; Testing; Time; Toxic effect; Transcriptional Regulation; Transfection; Transmission Electron Microscopy; Trauma; Ultraviolet Rays; Vertebrate Photoreceptors; Vision; Visual impairment; Zebrafish; Zinc; Zinc Fingers; age related; base; chromatin immunoprecipitation; cigarette smoking; comparative; disorder risk; environmental stressor; environmental tobacco smoke exposure; human disease; in vivo; in vivo imaging; insight; lens; lifestyle factors; novel; overexpression; promoter; public health relevance; research study; response; tool; toxic metal; transcription factor; transcription factor MTF-1; transcriptome sequencing; transcriptomics; tumor progression

 DESCRIPTION (provided by applicant): Eye pathologies, such as cataracts and macular degeneration, are the leading causes of impaired vision and blindness in the world. Although these pathologies form as a natural process of aging, there are several environmental and lifestyle factors that increase the risk of these diseases, such as cigarette smoking, exposure to light and ultraviolet radiation, oxidative stress, steroid treatment and stress-induced DNA damage. We propose to investigate the role of the metal-responsive transcription factor 1 (MTF-1) in the transcriptional regulation of a suite of genes involved in eye development that may also contribute to environmental stress- related eye pathologies. The main objectives of this proposal are to determine what role MTF-1 plays in normal eye development and whether or not inappropriate activation of MTF-1 signaling by cellular stress in the eye can lead to misexpression of genes that promote eye pathologies. Zebrafish embryos and adults will be used as our model organism and comparative experiments will be performed in a human primary lens epithelial cell line. Targeted gene expression assessments by in situ hybridization, real-time PCR and global transcriptomic profiling via RNA-seq will be used to determine the effects of MTF-1 activation or inhibition on the regulation of eye-specific genes. We have chosen the toxic metal cadmium as our model environmental risk factor because it is one of the best known activators of MTF-1 signaling and because it is believed to be a direct causative factor in the increased risk of cigarette smokers to cataractogenesis and macular degeneration. ChIP-Seq analysis complementary to the microarray expression profiling will be used to determine DNA binding by MTF- 1 as confirmation of a direct role in the regulation of genes responsible for eye development and stress-related pathologies. Additional promoter characterization via transient transfection assays and chromatin immunoprecipitation will be used for final confirmation of MTF-1 regulation of the eye-specific genes in zebrafish and humans. The results obtained from these experiments will yield public health significance by providing novel insight into mechanisms of action by which environmental stressors contribute to eye pathologies. In addition to the discovery of a potential new regulatory pathway involved in eye development with new insight into cataractogenesis and macular degeneration, successful completion of the proposed research will further expand the current use of zebrafish as a model organism for eye disease-related research.

 描述（由申请人提供）：白内障和黄斑变性等眼部疾病是世界范围内视力受损和失明的主要原因，尽管这些疾病是衰老的自然过程，但有一些环境和生活方式因素会加剧这些疾病。这些疾病的风险，如吸烟、暴露于光和紫外线辐射、氧化应激、类固醇治疗和应激诱导的 DNA 损伤，我们建议研究金属响应转录因子的作用。 MTF-1 在参与眼睛发育的一系列基因的转录调节中发挥作用，这些基因也可能导致与环境压力相关的眼睛病理。该提案的主要目的是确定 MTF-1 在正常眼睛发育中发挥的作用。眼睛细胞应激对 MTF-1 信号的不适当激活是否会导致促进眼睛病变的基因错误表达，我们将使用斑马鱼胚胎和成体作为我们的模型生物，并在人类初级晶状体中进行比较实验。上皮细胞系。通过原位杂交、实时 PCR 和通过 RNA-seq 进行的全局转录组分析进行的靶向基因表达评估将用于确定 MTF-1 激活或抑制对眼睛特异性基因调节的影响。我们将镉作为我们的模型环境风险因素，因为它是最著名的 MTF-1 信号激活剂之一，并且被认为是吸烟者白内障发生和黄斑变性风险增加的直接致病因素。与微阵列表达谱互补的 ChIP-Seq 分析将用于确定 MTF-1 的 DNA 结合，以确认在负责眼睛发育和应激相关病理的基因调节中的直接作用，并通过瞬时转染测定和其他启动子表征。染色质免疫沉淀将用于最终确认 MTF-1 对斑马鱼和人类眼睛特异性基因的调节。这些实验获得的结果将通过提供对作用机制的新见解而产生公共卫生意义。环境压力因素有助于眼部病理学的发现，除了发现了参与眼部发育的潜在新调控途径以及对白内障发生和黄斑变性的新见解外，拟议研究的成功完成还将进一步扩大斑马鱼作为模型生物的当前用途。眼病相关研究。