Evaluation of Endothelial Hyperglycemia-Driven Alterations During Type 2 Diabetes

2 型糖尿病期间内皮高血糖驱动变化的评估

• 批准号: 9145729
• 负责人: Brian Robert Hoffmann
• 金额: $ 10.13万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-17 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9145729
• 关键词: Accounting; Acetylcholine; Affect; Agonist; Area; Automobile Driving; Biochemical; Bioinformatics; Biological Assay; Biology; Blood Vessels; Cardiovascular Diseases; Cell physiology; Cell surface; Clinical; Complex; Coronary Arteriosclerosis; Coupled; Data; Development; Diabetes Mellitus; Diagnosis; Educational workshop; Endothelial Cells; Endothelium; Ensure; Epidemic; Equilibrium; Evaluation; Exposure to; Fostering; Functional disorder; Funding; Gene Expression; Glucose; Glycoproteins; HTR2A gene; Health; Homology Modeling; Human; Hyperglycemia; Inflammatory; Ions; K-Series Research Career Programs; Lead; Mass Spectrum Analysis; Measures; Medicine; Membrane; Membrane Glycoproteins; Mentors; Modeling; Nitric Oxide; Non-Insulin-Dependent Diabetes Mellitus; Ontology; Outcome; Outcomes Research; Pathway interactions; Patients; Peripheral Vascular Diseases; Pharmacology; Phenotype; Phosphorylation; Physiological; Physiology; Principal Investigator; Process; Production; Prostaglandins; Protein Glycosylation; Proteomics; Protocols documentation; Rattus; Receptor, Angiotensin, Type 1; Regulation; Research; Research Design; Research Personnel; Research Proposals; Risk; Sequence Homology; Signal Pathway; Signal Transduction; Site; Surface; System; Techniques; Technology; Testing; Therapeutic Intervention; Time; Training; Tube; United States; Validation; Vascular Diseases; Vascular Endothelium; Vascular System; base; career development; clinically relevant; cohort; design; diabetic patient; glycosylation; improved; innovation; medical specialties; mid-career faculty; mortality; novel; professor; programs; protein expression; receptor; research study; response; success

 DESCRIPTION (provided by applicant): Diabetes currently afflicts approximately 25.8 million people in the United States (US) and 220 million people worldwide. Type 2 diabetes mellitus (T2DM) accounts for ~95% of all diagnosed cases in the US, among which cardiovascular disease is the leading cause of mortality. Vascular dysfunction, leading to increased risk of coronary artery disease and peripheral vascular disease, remains an important clinical problem in diabetic patients. Hyperglycemia is a major causative factor of T2DM contributing to vascular dysfunction, however, there is a large gap of mechanistic studies in this area. This proposal evaluates the underlying hyperglycemia- induced alterations in the endothelium glycoproteome that lead to changes in important homeostatic signaling pathways, such as the AT1R and HTR2A signaling pathways, and result in vascular dysfunction. The aims of this proposal utilize innovative glycoproteome and phosphoproteome enrichment, sensitive mass spectrometry technology, and physiologically relevant functional tests to explore hyperglycemia-induced protein glycosylation at the cell surface (Aim 1), alterations of the synergistic balance of intracellular O-GlcNAcylation and phosphorylation (Aim 2), and subsequent alterations in important homeostatic signaling pathways leading to vascular dysfunction (Aim 3). In order to do this, the applicant requires protected time for supervised career development in bioinformatics, proteomics as applied to physiological systems, vascular biology, clinically relevant human cohort experimentation, and professional development under the direction of Dr. Andrew Greene (Professor of Physiology-MCW) and co-mentor Dr. Michael Widlansky (Associate Professor of Medicine and Pharmacology-MCW). This research proposal and career development modules will assist in fostering the Principal Investigator in his independence such that he can lead a research program aimed at examining causal factors in T2DM. The mentored career development award will further the candidate's training in bioinformatics processing for signal pathway ontology analyses, along with continued training in proteomics analysis applied to complex systems. The applicant will also receive extensive training in vascular biology as it relates to functional studies of T2DM rat models and vessels from T2DM patients. The training agenda includes lab-based training (MCW), clinical training (MCW/Froedtert & CTSI), specialty training in external labs, coursework, and professional development seminars/workshops. This multi-disciplinary training will ensure the ability of the applicant to design, perform, troubleshot, and interpret experiments independently. Outcomes of research proposal described here will provide direct evidence to confirm or refute the fundamental hypothesis that hyperglycemia-induced glycosylation is driving specific alterations leading to vascular dysfunction in T2DM and in combination with the professional career development will foster a smooth transition of the candidate to an independently funded investigator.

 描述（由申请人提供）：目前，美国约有 2580 万人患有糖尿病，全球约有 2.2 亿人患有 2 型糖尿病 (T2DM)，约占美国所有诊断病例的 95%，其中心血管疾病。血管功能障碍是导致冠状动脉疾病和周围血管疾病风险增加的主要原因，仍然是糖尿病患者的重要临床问题。高血糖是导致血管功能障碍的 T2DM 的一个主要致病因素，然而，该领域的机制研究还存在很大差距，这需要评估高血糖引起的内皮糖蛋白组的变化，从而导致重要的稳态信号通路的变化。作为 AT1R 和 HTR2A 信号通路，并导致血管功能障碍 该提案的目的是利用创新的糖蛋白质组和磷酸蛋白质组富集，灵敏的质谱技术和生理相关的功能测试，以探索高血糖诱导的细胞表面蛋白质糖基化（目标 1）、细胞内 O-GlcNA 酰化和磷酸化协同平衡的改变（目标 2）以及重要稳态信号传导的后续改变导致血管功能障碍的途径（目标 3）为了做到这一点，申请人需要有受保护的时间来监督生物信息学的职业发展，在 Andrew Greene 博士（MCW 生理学教授）和共同导师 Michael Widlansky 博士（医学和药理学副教授）的指导下，蛋白质组学应用于生理系统、血管生物学、临床相关的人类队列实验和专业发展MCW）。该研究提案和职业发展模块将有助于培养首席研究员的独立性，以便他能够领导一项旨在研究 T2DM 因果因素的研究计划。将进一步对候选人进行信号通路本体分析的生物信息学处理培训，以及应用于复杂系统的蛋白质组学分析的继续培训。申请人还将接受血管生物学方面的广泛培训，因为它涉及 T2DM 大鼠模型和 T2DM 血管的功能研究。培训议程包括实验室培训 (MCW)、临床培训（MCW/Froedtert 和 CTSI）、外部实验室专业培训、课程和专业发展研讨会/讲习班。多学科培训将确保申请人独立设计、执行、排除故障和解释实验的能力。此处描述的研究计划的结果将提供直接证据来证实或反驳高血糖诱导的糖基化正在驱动导致特定改变的基本假设。对 T2DM 血管功能障碍的研究，并与职业生涯发展相结合，将促进候选人顺利过渡为独立资助的研究者。