Project 1

心血管疾病是糖尿病患者死亡的主要原因，并且可能在没有其他已知风险因素的情况下发生。线粒体生物能量缺陷和自由基产生增加是糖尿病心肌病（DCM）的病理标志。该项目的一个目标是确定线粒体中发生的分子变化，以诱导代谢功能障碍和氧化应激。具体来说，我们正在解决如何失调线粒体蛋白赖氨酸乙酰化 导致代谢紊乱、线粒体功能障碍和DCM的进展。我们的假设是，高血糖导致特定代谢酶（如蛋白激酶A）的病理性蛋白质赖氨酸乙酰化增加，这有助于脂肪酸氧化、线粒体功能障碍和氧化应激增加。使用1型糖尿病的转基因啮齿动物模型和细胞培养技术，我们将测试假设如下：目的1。定义发生的更改， 糖尿病患者体内的氧化应激反应与线粒体有关，导致线粒体功能障碍，并随着糖尿病的进展而增加氧化应激。线粒体底物选择、氧化磷酸化和自由基产生将与心脏结构和功能的变化同时使用磁共振成像（MRI）和组织学进行分析。目标二。定义超乙酰化对线粒体功能障碍的贡献。的 将确定超乙酰化对氧化磷酸化、氧化应激和糖尿病性心肌病的影响。这将通过a）乙酰化蛋白质的MS分析; B）鉴定高乙酰化诱导的功能变化;和c）鉴定糖尿病诱导的高乙酰化的原因来完成。目标3.确定PKA在促进代谢稳定性和线粒体功能障碍中的作用。这一目标将检验我们观察到的PKA活性降低是一个假设。 通过氧化和/或乙酰化介导。机制研究将确定这些修饰对线粒体呼吸活性和自由基产生的发生和后果。

Cardiovascular disease is the leading cause of death among people with diabetes and may occur in the absence of other known risk factors. Mitochondrial bioenergetic deficits and increased free radical production are pathological hallmarks of diabetic cardiomyopathy (DCM). A goal of this project is to determine the molecular changes that occur in mitochondria to induce metabolic dysfunction and oxidative stress. Specifically, we are addressing how dysregulated mitochondrial protein lysine acetylation contributes to metabolic inflexibility, mitochondrial dysfunction, and the progression of DCM. Our hypothesis is that hyperglycemia leads to increased, pathological protein lysine acetylation of specific metabolic enzymes, such as protein kinase A, and this contributes to fatty acid oxidation, mitochondrial dysfunction, and increased oxidative stress. Using a transgenic rodent model of type 1 diabetes and cell culture techniques, we will test the hypothesis as follows: Aim 1. Define the changes that occur to mitochondria that lead to mitochondrial dysfunction and increased oxidative stress with the progression of diabetes. Mitochondrial substrate selection, oxidative phosphorylation, and free radical production will be analyzed in parallel with changes in cardiac structure and function using magnetic resonance imaging (MRI) and histology. Aim 2. Define the contribution of hyper-acetylation to mitochondrial dysfunction. The consequences of hyper-acetylation on oxidative phosphorylation, oxidative stress, and diabetic cardiomyopathy will be determined. This will be done by a) MS analysis of acetylated proteins; b) identifying the functional changes that hyperacetylation induces; and c) identifying the cause of diabetes induced hyperacetylation. Aim 3. Determine the role of PKA in contributing to metabolic inflexibility and mitochondrial dysfunction. This aim will test the hypothesis that our observed decrease in PKA activity is mediated by oxidation and/or acetylation. Mechanistic studies will determine the occurrence and consequences of these modifications on mitochondrial respiratory activity and free radical production.