Dual Role of HSP70 in Diabetes-Induced Vascular Dysfunction

HSP70 在糖尿病引起的血管功能障碍中的双重作用

• 批准号: 10515009
• 负责人: Kenia Pedrosa Nunes
• 金额: $ 43.42万
• 依托单位: FLORIDA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-04 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10515009
• 关键词: Address; Affect; Agonist; Animals; Antibodies; Aorta; Atomic Force Microscopy; Automobile Driving; Biology; Biomedical Engineering; Blood Vessels; Cell Adhesion; Cell Compartmentation; Cells; Chronology; Collaborations; Confocal Microscopy; Cultured Cells; Data; Deposition; Diabetes Mellitus; Disease; Engineering; Environment; Enzyme-Linked Immunosorbent Assay; Exposure to; Extracellular Matrix; Female; Florida; Glucose; Health; Heat-Shock Proteins 70; Human; Hyperglycemia; ITPR1 gene; Immune system; Immunoglobulin G; In Vitro; Inflammation; Interleukin-1 beta; Interleukin-6; Laboratories; Link; Literature; Measures; Mesentery; Methods; Molecular; Molecular Chaperones; Mus; NADPH Oxidase; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Outcomes Research; Oxidative Stress; Pharmaceutical Preparations; Pharmacology; Physiologic pulse; Physiological; Play; Proteins; Research; Research Training; Rho-associated kinase; Role; Science; Serum; Severities; Signal Transduction; Small Interfering RNA; Smooth Muscle; Smooth Muscle Myocytes; Source; Students; Supervision; TNF gene; Techniques; Testing; Thermogenesis; Tissues; Transforming Growth Factor beta; Ultrasonography; Up-Regulation; Vascular Diseases; Vascular Smooth Muscle; Western Blotting; arterial stiffness; base; cytokine; db/db mouse; diabetes management; experience; experimental study; extracellular; in vivo; inhibitor; innovation; knock-down; male; mouse model; multidisciplinary; nanoscale; prevent; professor; response; sex; ultrasound; undergraduate student

Dual role of HSP70 in diabetes-induced vascular dysfunction R15 – Applicant Kenia Nunes Project Summary/Abstract Vascular damage, a significant health problem, is linked to hyperglycemia, a key component of both types of diabetes. The rationale for this study is that increased glucose levels chronologically associate with higher levels of circulating Heat-shock protein 70 (HSP70), an intriguing molecular chaperone that co-exists in different cell compartments and exerts antagonistic actions. There is a well-known crosstalk between HSP70 and Ca2+, but not in smooth muscle. Only recently, my lab showed that a functional iHSP70 is required for adequate vascular reactivity under physiological conditions, as it impacts Ca2+ handling mechanisms. In diabetes, there is an increase in the levels of eHSP70, which is associated with oxidative stress and low-grade inflammation. Thus, it is reasonable to argue that HSP70 could determine the outcome of vascular damage in this disease. Still, it is unknown if iHSP70 affects vascular Ca2+ signaling under hyperglycemic conditions and if eHSP70 contributes to vascular functional and structural alterations in diabetes. The central hypothesis of this project, formulated based on the literature and my robust preliminary data, is that HSP70 plays a dual role in diabetes-induced vascular dysfunction. This study is innovative because it will independently investigate the interplay between iHSP70 and Ca2+ handling mechanisms leading to functional and structural changes under hyperglycemia and by selectively targeting eHSP70 in a murine model of type 2 diabetes to prevent these changes. I will combine a set of well-established in vitro, ex vivo, and in vivo approaches, by using state-of-art techniques such as atomic force microcopy and ultrasonography (Pulse Wave Velocity) to allow observations at macro, micro and nano scale. Also, sex plays a crucial role in determining the tissue rate of production of HSP70; therefore, we will use male and female animals. The experiments outlined in this study will address each specific aim efficiently by utilizing the strengths of my laboratory. Moreover, network between biology and engineering will be stablished by collaboration with a senior professor in Biomedical Engineering. The PI has extensive experience in the field and will closely supervise the students. The outcome of this research has an enormous potential to uncover target mechanisms to manage diabetic vasculopathies while creating a multidisciplinary environment offering extensive research training to undergraduate students. Our findings will have a broader impact by opening new research avenues for other diseased states associated with vascular structural and functional changes.

HSP70 在糖尿病引起的血管功能障碍中的双重作用 R15 – 申请人 Kenia Nunes 项目概要/摘要 血管损伤是一个严重的健康问题，与高血糖有关，而高血糖是糖尿病的一个关键组成部分。 两种类型的糖尿病。这项研究的基本原理是，葡萄糖水平按时间顺序增加 与较高水平的循环热休克蛋白 70 (HSP70) 相关，这是一种有趣的分子 分子伴侣共存于不同的细胞区室中并发挥拮抗作用。有 众所周知，HSP70 和 Ca2+ 之间存在串扰，但平滑肌中却没有这种串扰。直到最近，我的 实验室表明，功能性 iHSP70 是在条件下足够的血管反应性所必需的 生理条件，因为它影响 Ca2+ 处理机制。在糖尿病中，有一个 eHSP70 水平增加，这与氧化应激和低级别相关 炎。因此，有理由认为 HSP70 可以决定血管的结果。 这种疾病的损害。尽管如此，目前尚不清楚 iHSP70 是否会影响血管 Ca2+ 信号传导 高血糖状况以及 eHSP70 是否有助于血管功能和结构 糖尿病的改变。该项目的中心假设是根据文献制定的 我强有力的初步数据是，HSP70 在糖尿病诱发的血管生成中发挥双重作用 功能障碍。这项研究具有创新性，因为它将独立调查相互作用 iHSP70 和 Ca2+ 处理机制之间导致功能和结构变化 在高血糖下并通过在 2 型糖尿病小鼠模型中选择性靶向 eHSP70 以防止这些变化。我将结合一套成熟的体外、离体和体内 方法，通过使用最先进的技术，如原子力显微镜和 超声波检查（脉冲波速度）可在宏观、微观和纳米尺度上进行观察。 此外，性别在决定 HSP70 的组织生成率方面也起着至关重要的作用。因此，我们 将使用雄性和雌性动物。本研究中概述的实验将解决每个特定的问题 利用我实验室的优势有效地瞄准目标。此外，生物学和 工程将通过与生物医学工程高级教授合作建立。 PI在该领域拥有丰富的经验，并将密切监督学生。结果 这项研究具有揭示治疗糖尿病的目标机制的巨大潜力 血管疾病，同时创建提供广泛研究培训的多学科环境 给本科生。我们的发现将通过开展新的研究产生更广泛的影响 与血管结构和功能变化相关的其他疾病状态的途径。