Transforming Growth Factor Beta-Activated Kinase 1 (Tak1) in Retinal Microglial Inflammation

转化生长因子 β 激活激酶 1 (Tak1) 在视网膜小胶质细胞炎症中的作用

• 批准号: 10438002
• 负责人: TERI L BELECKY-ADAMS
• 金额: $ 45.6万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10438002
• 关键词: Affect; Alzheimer' s Disease; Animal Model; Astrocytes; B-Cell Activation; Blood flow; Blood-Retinal Barrier; Brain; Brain region; Carbon Dioxide; Cell Count; Cells; Cerebrovascular Circulation; Chronic; Complement Factor B; Complication; Critical Pathways; Data; Diabetes Mellitus; Diabetic Retinopathy; Disease; Elements; Endothelial Cells; Enhancers; Ensure; Enzyme-Linked Immunosorbent Assay; Failure; Genetic Transcription; Goals; HMGB1 Protein; Health; Heterogeneity; IL6 gene; Immune; Immunologic Surveillance; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Injury; Interferon Type II; Interleukin-6; Knock-out; Laboratories; Lead; Ligands; Light; MAP3K7 gene; MAPK8 gene; Measurement; Measures; Mediating; Messenger RNA; Metabolic; Microglia; Microinjections; Muller' s cell; Mus; Neuraxis; Neurodegenerative Disorders; Neurons; Neuropil; Nuclear; Nutrient; Oxygen; Parkinson Disease; Pathway interactions; Pericytes; Pharmacologic Substance; Phosphotransferases; Play; Polymerase Chain Reaction; Population; Positioning Attribute; Process; Proteins; Research Personnel; Retina; Retinal Diseases; Role; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Streptozocin; Supporting Cell; TNF gene; Testing; Transforming Growth Factor beta; Transforming Growth Factors; Type 2 diabetic; Up-Regulation; Western Blotting; astrogliosis; cohort; diabetic patient; experimental study; factor A; ganglion cell; glymphatic system; in vitro testing; in vivo; inhibitor; macroglia; member; neurovascular unit; new therapeutic target; p38 Mitogen Activated Protein Kinase; small molecule inhibitor; therapeutic target; type I diabetic; wasting

Diabetic retinopathy is associated with chronic aberrant inflammation that is proposed to play a critical role in the early disruption of neurovascular unit function. There is incomplete information about intracellular signaling pathways involved in upregulation of inflammatory signals. Identification of more pathway members will lead to more therapeutic targets to be used in treatment of neurodegenerative diseases. This proposal is aimed at testing the role of transforming growth factor β-activated kinase 1 (TAK1) in activation NFκB, p38, and JNK pathways in retinal microglia, resulting in increased transcription of inflammatory factors. The hypothesis that inhibition of TAK1 will reduce inflammation in the early stages of diabetic retinopathy and reduce changes to the neurovascular unit will be tested in vitro using isolated microglial cells and in vivo using a combination of streptozotocin (STZ)-induced diabetes in mice with conditional loss of TAK1 in microglia. Experiments will utilize small molecule inhibitors, addition of known inflammatory factors, microinjections, quantitative polymerase chain reactions, Western blot analysis, multiplex enzyme-linked immunosorbent assays, and quantitation of pericytes, microvasculature, microglia, retinal astrocytes, ganglion cells, and Müller glia in retinal wholemounts and sections.

糖尿病视网膜病变与慢性异常炎症有关，慢性异常炎症被认为在糖尿病视网膜病变中起关键作用。 神经血管单位功能的早期破坏。关于细胞内信号传导的信息不完整 参与炎症信号上调的途径。识别更多途径成员将导致 更多的治疗靶点用于治疗神经退行性疾病。这一建议旨在 检测转化生长因子β激活激酶1（TAK1）在NFκB、p38和JNK激活中的作用 视网膜小胶质细胞中的信号通路，导致炎症因子的转录增加。的假设 抑制TAK1将减少糖尿病视网膜病变早期的炎症， 神经血管单元将在体外使用分离的小胶质细胞进行测试， 链脲佐菌素（STZ）诱导的小鼠糖尿病伴小胶质细胞中TAK1的条件性丢失。实验将 利用小分子抑制剂，添加已知的炎症因子，显微注射，定量 聚合酶链反应、蛋白质印迹分析、多重酶联免疫吸附测定，以及 视网膜中周细胞、微血管、小胶质细胞、视网膜星形胶质细胞、神经节细胞和Müller胶质细胞的定量 整体和部分。