ShEEP Request for Intracellular and Extracellular Protein Signaling Station (IEPSS)

ShEEP 请求细胞内和细胞外蛋白质信号转导站 (IEPSS)

• 批准号: 10178936
• 负责人: Bruce R. Troen
• 金额: --
• 依托单位: VA WESTERN NEW YORK HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10178936
• 关键词: Acinetobacter baumannii; Acute; Age related macular degeneration; Aging; Antioxidants; Biological Markers; Cardiac; Catalytic RNA; Cell Death; Cell Separation; Cells; Charge; Chronic; Clinical Research; Collaborations; Collection; Cultured Cells; DNA Methylation; Data; Detection; Development; Devices; Disease; Epithelial Cells; Equipment; Extracellular Protein; Functional disorder; Funding; Future; Gel; Health; Heart Arrest; Hematopoietic; Hour; Human; Image; Immunoassay; Immunoblotting; Inflammation; Inherited; Interval training; Ischemia; Manufacturer Name; Measurement; Measures; Mediating; Membrane Proteins; Methodology; Methods; Minor; Mitochondria; Molecular; Molecular Weight; Multiple Trauma; Mus; Mutation; Myocardial Infarction; Orphan; Oxidative Stress; Parkinson Disease; Pathologic; Pathologic Processes; Penicillin-Binding Proteins; Performance; Photoreceptors; Physical Performance; Physiological; Post-Translational Protein Processing; Predictive Analytics; Preparation; Process; Protein Isoforms; Proteins; Proteomics; Quality of life; RNA Interference; Reproducibility; Reproducibility of Results; Research; Research Personnel; Research Project Grants; Retina; Retinal Degeneration; Retinal Dystrophy; Retinitis Pigmentosa; Role; Running; Sampling; Sheep; Signaling Protein; Skeletal Muscle; Therapeutic Agents; Therapeutic Intervention; Time; Tissues; Translational Research; Universities; Validation; Veterans; Western Blotting; Work; antimicrobial; base; bench to bedside; cardiac repair; chemokine; cost; cytokine; dopaminergic neuron; exosome; frailty; healthspan; human model; improved; insight; instrument; macrophage; military veteran; novel; parkin gene/protein; pre-clinical; pre-clinical research; prevent; programs; protein biomarkers; protein expression; receptor; repaired; response; single-cell RNA sequencing; skeletal muscle metabolism; stem cells; targeted treatment; therapeutic candidate; therapy development; visual dysfunction

Project Abstract The Bio-Techne Intracellular and Extracellular Protein Signaling Station (IEPSS) automates standard western blotting and immunoassay and produces reliable, reproducible quantitative data in a very short time. The IEPSS provides a wide array of capabilities, including single cell western blotting, identification of protein isoforms and post-translational modifications in a single cell, separation of proteins based on size and charge, measure four or more analytes in multiple samples. Thus, the IEPSS will provide immediate benefits for VAWNYHS investigators studying the underlying acute and chronic processes that result in physiological dysfunction, disease, and health decline, ultimately permitting the development of interventions to improve the quality of life in the Veteran population. These projects include: Dr. Troen’s research investigates physical performance, frailty, and healthspan during aging and is presently studying the benefits of high intensity interval training (HIIT) for skeletal muscle performance in aging veterans. The IEPSS will allow assessment of protein markers and posttranslational modifications associated with mitochondrial health and skeletal muscle metabolism. Dr. Fliesler’s projects investigate the underlying mechanism of progressive retinal degeneration and visual dysfunction associated with blast overpressure-induced polytrauma, and the impact of novel antioxidants as therapeutic agents to prevent, minimize, or slow the progression of the pathological processes. The IEPSS will afford assessment of biomarkers of inflammation, oxidative stress, and cell death. Dr. Sullivan’s study investigates the development of post-transcriptional gene silencing agents such as ribozymes as candidate therapeutics for orphan retinal degenerations and common age-related macular degeneration. The IEPSS will permit assessment of ribozyme rescue strategy and quantify target protein expression and post-translational modification in photoreceptor and retinal epithelial cells. Dr. Canty’s study investigates cellular remodeling during sudden cardiac arrest and cardiac stem cell mediated repair. The IEPSS will facilitate a proteomic profile of hematopoietic subtypes as well as macrophage subpopulations, also enable to study the chemokines/cytokines. In addition, it quantifies protein changes that arise in response to reversible ischemia. Dr. Feng’s study seeks to identify more efficient methods to understand how mutations of parkin cause the selective degeneration of human dopaminergic neurons and the ensuing Parkinson’s disease. The IEPSS will allow for characterization of protein expression profiles and posttranslational modifications. Dr. Russo’s research seeks to determine the role of penicillin binding protein (PBP) 7/8 as a novel anti- microbial target in XDR A. baumannii. The IEPSS will enable to resolve molecular mechanism of PBP 7/8 and develop receptor-targeted therapies by profiling various outer membrane proteins in single and cultured cells, identifying post-translational modifications in the presence or absence of PBP 7/8. Dr. Farkas’ research seeks to discover potential therapies to prevent and/or treat age-related macular degeneration (AMD) and inherited retinal dystrophies (IRDs). The IEPSS will enable validation of the effects of DNA methylation changes on corresponding protein expression and quantify the cell-to-cell variability of the retinitis pigmentosa 1 minor isoform in the mouse retina. Dr. Lang’s research seeks to determine the functional role of exosomes in stem cell-mediated cardiac repair. The IEPSS will enhance this work by performing multiple immunoassays on small quantities of initial sample and providing the ability to quantify cytokine panels in response to exosome therapy and measuring protein expression in stem cells and in murine cardiac tissue following myocardial infarction.

项目摘要 Bio-Techne 细胞内和细胞外蛋白质信号转导站 (IEPSS) 实现自动化 标准的蛋白质印迹和免疫测定，并以非常精确的方式产生可靠、可重复的定量数据 短时间。 IEPSS 提供广泛的功能，包括单细胞蛋白质印迹、 单细胞中蛋白质亚型和翻译后修饰的鉴定、蛋白质的分离 根据尺寸和电荷，测量多个样品中的四种或多种分析物。因此，IEPSS 将提供 VAWNYHS 研究人员研究潜在的急性和慢性过程的直接好处 导致生理功能障碍、疾病和健康状况下降，最终导致 改善退伍军人生活质量的干预措施。这些项目包括： Troen 博士的研究调查了衰老过程中的身体表现、虚弱程度和健康寿命， 目前正在研究高强度间歇训练（HIIT）对骨骼肌表现的好处 年迈的退伍军人。 IEPSS 将允许评估蛋白质标记和翻译后修饰 与线粒体健康和骨骼肌代谢有关。 Fliesler 博士的项目研究了进行性视网膜变性的潜在机制和 与爆炸超压引起的多发伤相关的视觉功能障碍，以及新型药物的影响 抗氧化剂作为治疗剂来预防、最小化或减缓病理进展 流程。 IEPSS 将提供炎症、氧化应激和细胞死亡生物标志物的评估。 沙利文博士的研究调查了转录后基因沉默剂的开发，例如 作为核酶作为孤儿视网膜变性和常见年龄相关性黄斑的候选疗法 退化。 IEPSS 将允许评估核酶救援策略并量化目标蛋白 光感受器和视网膜上皮细胞中的表达和翻译后修饰。 Canty 博士的研究调查了心脏骤停期间的细胞重塑和心脏干细胞 介导修复。 IEPSS 将促进造血亚型的蛋白质组学谱以及 巨噬细胞亚群也能够研究趋化因子/细胞因子。此外，它还可以定量蛋白质 因可逆性缺血而发生的变化。 冯博士的研究旨在寻找更有效的方法来了解 Parkin 突变如何发生 导致人类多巴胺能神经元的选择性变性以及随后的帕金森病。这 IEPSS 将允许对蛋白质表达谱和翻译后修饰进行表征。 Russo 博士的研究旨在确定青霉素结合蛋白 (PBP) 7/8 作为新型抗 XDR 鲍曼不动杆菌中的微生物靶标。 IEPSS将能够解析PBP 7/8的分子机制和 通过分析单一和培养的各种外膜蛋白来开发受体靶向疗法 细胞，识别存在或不存在 PBP 7/8 的情况下的翻译后修饰。 Farkas 博士的研究旨在发现预防和/或治疗年龄相关性黄斑的潜在疗法 变性（AMD）和遗传性视网膜营养不良（IRD）。 IEPSS 将验证效果 DNA 甲基化变化对相应蛋白质表达的影响，并量化细胞间的变异性 小鼠视网膜中的色素性视网膜炎 1 小亚型。 Lang 博士的研究旨在确定外泌体在干细胞介导的心脏功能中的功能作用 维修。 IEPSS 将通过对少量初始样本进行多次免疫测定来加强这项工作。 样本并提供量化细胞因子组以响应外泌体治疗和测量的能力 心肌梗塞后干细胞和小鼠心脏组织中的蛋白质表达。