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传导站（IEPSS） 标准蛋白质印迹和免疫测定，并产生可靠的，可重复的定量数据，在一个非常 短时间IEPSS提供了广泛的功能，包括单细胞免疫印迹， 在单个细胞中鉴定蛋白质同种型和翻译后修饰，分离蛋白质 基于大小和电荷，测量多个样品中的四种或更多种分析物。因此，IEPS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，疾病和健康下降，最终允许发展 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、虚弱和健康状况， 目前正在研究高强度间歇训练（HIIT）对骨骼肌性能的益处， 上了年纪的老兵IEPSS将允许评估蛋白质标记物和翻译后修饰 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， 与冲击波超压引起的多发伤相关的视觉功能障碍，以及新的 抗氧化剂作为治疗剂以预防、最小化或减缓病理性 流程. IEPSS将评估炎症、氧化应激和细胞死亡的生物标志物。 沙利文博士的研究调查了转录后基因沉默剂的发展， 作为孤儿视网膜变性和常见年龄相关性黄斑变性的候选治疗剂的核酶 退化IEPSS将允许评估核酶拯救策略并定量靶蛋白 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， 巨噬细胞亚群也使得能够研究趋化因子/细胞因子。此外，它还能定量蛋白质 可逆性缺血引起的变化。 冯博士的研究旨在确定更有效的方法来了解parkin基因的突变是如何发生的。 导致人类多巴胺能神经元的选择性变性和随之而来的帕金森病。的 IEPSS将允许表征蛋白质表达谱和翻译后修饰。 Russo博士的研究旨在确定青霉素结合蛋白（PBP）7/8作为一种新的抗- XDR A中的微生物靶标。鲍曼不动杆菌。IEPSS将能够解决PBP 7/8的分子机制， 通过在单个和培养的细胞中分析各种外膜蛋白， 细胞，在存在或不存在PBP 7/8的情况下鉴定翻译后修饰。 Farkas博士的研究旨在发现潜在的治疗方法，以预防和/或治疗年龄相关性黄斑病变。 视网膜变性（AMD）和遗传性视网膜营养不良（IRD）。IEPSS将使效果得到验证 DNA甲基化改变对相应蛋白质表达的影响，并量化 视网膜色素变性1小亚型在小鼠视网膜。 Lang博士的研究旨在确定外泌体在干细胞介导的心脏疾病中的功能作用。 修复. IEPSS将通过对少量初始免疫原性细胞进行多种免疫测定来加强这项工作。 样品，并提供响应于外来体疗法定量细胞因子组的能力， 心肌梗死后干细胞和鼠心脏组织中的蛋白表达。