Leveraging Protein Kinase G-1 Nanodomain Control and Molecular Targeting to Enhance its Therapeutic Use Against Myocardial Disease

这个项目的目标是重新发明关于监管、目标确定和 特别是蛋白激酶G(PKG)激活的翻译用途，以达到治疗的目的 心肌病。PKG是由第二信使环GMP(CGMP)激活的主要酶 也是血管紧张度的重要调节者。它在心肌细胞中的作用更多 有争议，但越来越多的证据表明，刺激PKG提供了一种有效的抗压力和 病理生理刹车，对抗促肥大/纤维化信号，机械传感和 心律失常，改善舒张期功能。我们实验室的新数据现在表明，它也可以 深刻影响microRNA的形成、蛋白质质量控制和自噬。环状GMP是 由一氧化氮或利钠肽信号通路产生。虽然两者都有很长的时间 被认为是可互换的，我们最近的工作显示出它们在角色和 心肌细胞的调节。它们在受特定磷酸调节的纳米结构域中工作。 特别是以NO和NP为靶标的PDE5和PDE9分别刺激cGMP。因此，如何 一个人有效地激活PK取决于疾病的情况，因此cGMP是如何被激活的 产生，涉及哪些PDE，甚至PKG的翻译后状态。例如, 氧化应激抑制NO刺激的cGMP也氧化PKG，我们发现它减少了它的 保护作用，同时也改变其对激动剂的反应。雌激素耗竭损害 NO在雌性体内刺激cGMP，损害依赖于这一途径的PKG激活策略。 该R35计划开发了四个创新研究计划，旨在最终改善我们的 PKG激活的治疗用途：1)剖析纳米结构域控制，定义蛋白质伙伴， 选择性蛋白酪氨酸激酶靶点，它们在不同疾病中的动态，以及它们如何可以更多 有效监管；2)确定肥胖、代谢综合征和 限制绝经后PKG的激活策略，并制定规避策略；3) 发现PKG激活提供好处的新信号，包括在 自噬、蛋白小体运输、病理机械传感和转录调控；以及4) 使用新的蛋白质组学方法开发应用于人类心脏活检和血液的PKG蛋白质组型。 通过这项工作，我们的目标是转变PKG治疗心脏病的理念，基于其 生物学衍生出一种有效的个性化方法。

The goal of this project is to re-invent concepts regarding the regulation, targeting, and particularly the translational use of protein kinase G (PKG) activation for the goal of treating myocardial disease. PKG is the primary enzyme activated by the second messenger cyclic GMP (cGMP) and a prominent regulator of vascular tone. Its role in the cardiomyocyte has been more controversial, but growing evidence shows that stimulating PKG provides a potent anti-stress and pathophysiological brake, countering pro- hypertrophic/fibrotic signaling, mechano-sensing and arrhythmia, and improving diastolic function. New data from our lab now shows it can also profoundly impact microRNA formation, protein quality control, and autophagy. Cyclic GMP is generated by either a nitric oxide or natriuretic peptide signaling-pathway. While both have long been viewed as interchangeable, our recent work shows prominent differences in their role and regulation in the cardiomyocyte. They operate in nano-domains regulated by specific phospho- diesterases; notably PDE5 and PDE9 that target NO and NP stimulated cGMP, respectively. Thus, how one activates PK effectively depends upon the disease condition and thus how cGMP is being generated, which PDEs are involved, and even the post-translational state of PKG. For example, oxidative stress, which depresses NO-stimulated cGMP also oxidizes PKG, which we showed reduces its protective effects while also altering its response to agonists. Estrogen depletion impairs NO-stimulated cGMP in females, compromising PKG activation strategies dependent on this pathway. This R35 program develops four innovative research programs aimed at ultimately improving our therapeutic use of PKG activation: 1) Dissect nano-domain controls, defining protein partners, selective PKG kinase targets, their dynamics in varying diseases, and how they can be more effectively regulated; 2) Identify how co-morbidities such as obesity, metabolic syndrome, and post- menopause limit PKG activation strategies, and develop methods to circumvent them; 3) Discover novel signaling by which PKG activation provides benefit, including new roles in autophagy, proteosome trafficking, pathological mechanosensing and transcriptional controls; and 4) Develop a PKG proteotype using new proteomic methods applied to human heart biopsies and blood. Through this work, we aim to transform concepts of PKG therapy for heart disease, based on its biology to derive an effective personalized approach.