cGMP-dependent protein kinase I as a new target against stroke

cGMP依赖性蛋白激酶I作为抗中风的新靶点

• 批准号: 9240123
• 负责人: DMITRIY ATOCHIN
• 金额: $ 35.4万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9240123
• 关键词: Affect; Attenuated; Blood Platelets; Blood Vessels; Cerebral Ischemia; Cerebrovascular Circulation; Cerebrovascular system; Cerebrum; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Data; Development; Foundations; Functional disorder; Goals; Homeostasis; Infarction; Injury; Ischemia; Knock-out; Knockout Mice; Knowledge; Leukocytes; Link; Mediating; Mediator of activation protein; Molecular; Mus; Neurologic; Outcome; Patients; Phosphotransferases; Platelet aggregation; Regulation; Reperfusion Injury; Reperfusion Therapy; Role; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Soluble Guanylate Cyclase; Stroke; Testing; Therapeutic; Therapeutic Agents; Vasodilation; Wild Type Mouse; base; cGMP-dependent protein kinase I; cerebral microvasculature; cerebrovascular; middle cerebral artery; mouse model; protective effect; research study; targeted treatment

The signaling pathways by which NO protects against stroke injury are mediated by soluble guanylyl cyclase activation and cyclic guanosine monophosphate ( cGMP) formation. However, the mechanisms of downstream signaling from cGMP to smooth muscle cells (SMC) remain incompletely understood. The cGMP- dependent protein kinase I (cGKI) is a key mediator of cGMP signaling. Our preliminary results indicate that reperfusion of middle cerebral artery was attenuated and stroke injury was more pronounced in cGKI SMC specific inducible conditional knockout mice than in wild-type mice, suggesting a protective role for cGKI in SMC against cerebral reperfusion injury. The overall goal of this project is to identify the molecular mechanisms underlying the protective effect of cGKI signaling in SMC against stroke injury. We will test the hypotheses that cGKI mediate the protective effects of cGMP and that the protective effects of cGMP-cGKI signaling are SMC specific. We will take advantage of mice with inducible knockout of cGKI in SMC to specifically study the contributions of SMC-dependent vascular mechanisms to the protective effects of cGMP against stroke. Our goals are (1) to determine the role of cGKI SMC-dependent mechanisms to the protective effect in stroke and (2) to identify the molecular mechanisms underlying the protective role of cGKI-SMC signaling in stroke injury. These results will make possible the development of new specific therapeutic approaches to stroke. HypothesiscGKI in smooth muscle cells protects against stroke. Aim 1: To determine the relevance and mechanism of cGKI-mediated signaling in SMC for stroke protection. We will study whether deficiency of cGKI in SMC worsens stroke outcome in cGKI SMC- specific knockout mice. We will compare stroke outcome in cGKI knockout mice and cGKI wild-type mice. We will determine neurological deficit and infarct volume after occlusion and reperfusion of the middle cerebral artery and compare cerebral blood flow at baseline and during ischemia and reperfusion. Aim 2: To study the role of cGKI of SMC in vascular reactivity. We will test whether cGKI deletion in SMC impairs reactivity of isolated vessels, cerebral blood flow and autoregulation. Aim 3: To investigate the role of platelets and leukocytes in stroke injury of cGKI SMC deficient mice. Platelets aggregation and accumulation of leukocytes within cerebral microvessels is critically involved in stroke injury. We will study how cGKI deficiency in SMC affects the role of platelet aggregation and leukocytes accumulation in stroke injury. We anticipate that studies described in this proposal will establish a new mouse model for stroke studies and further our knowledge on the role of cGKI SMC signaling in the protection against stroke. These results will be critical to provide the scientific foundation for the development of cGKI-elevating compounds as therapeutic agents in stroke patients.

一氧化氮保护卒中损伤的信号通路是由可溶性鸟苷酸介导的 环化酶激活和环鸟苷一磷酸( (CGMP) 队形。然而，它的作用机制 从cGMP到平滑肌细胞(SMC)的下游信号仍然不完全清楚。CGMP- 依赖蛋白激酶I(CGKI)是cGMP信号转导的关键介质。我们的初步结果表明 CGKI-SMC组大鼠大脑中动脉再灌注量减少，卒中损伤更明显 特异性可诱导条件性基因敲除小鼠比野生型小鼠，提示cGKI在 SMC抗脑再灌注损伤。 这个项目的总体目标是确定潜在的保护作用的分子机制。 CGKI信号在SMC抗中风损伤中的作用。我们将检验cGKI介导保护性免疫的假设 CGMP-cGKI信号的保护作用是SMC特异性的。 我们将利用具有诱导性的老鼠 CGKI基因敲除 SMC 要专门研究 SMC依赖的血管机制在cGMP预防中风中的作用。 我们的 目的是(1)确定cGKI-SMC依赖的机制在卒中保护作用中的作用。 (2)确定潜在的分子机制 CGKI-SMC信号的保护作用 在中风中受伤。 这些结果将使开发治疗中风的新的特定治疗方法成为可能。 假设在平滑肌细胞中的cGKI可以预防中风。 目的1：确定cGKI介导的信号转导在SMC中的相关性和机制。 中风保护。我们将研究SMC中cGKI缺乏是否会恶化cGKI SMC的卒中结局。 特定的基因敲除小鼠。我们将比较cGKI基因敲除小鼠和cGKI野生型小鼠的中风结果。我们 将确定大脑中动脉闭塞和再灌流后的神经功能缺陷和脑梗塞体积 并比较基线和缺血再灌注期的脑血流量。 目的：研究SMC中cGKI在血管反应性中的作用。我们将测试cGKI是否删除 血管紧张素转换酶抑制分离血管的反应性、脑血流量和自我调节。 目的3：探讨血小板和白细胞在cGKI-SMC缺乏性卒中损伤中的作用 老鼠。血小板聚集和白细胞在脑微血管内聚集在脑微血管中起关键作用 中风伤。我们将研究SMC中cGKI缺乏如何影响血小板聚集和白细胞的作用 在中风损伤中积聚。 我们预计，这项提案中描述的研究将建立一种新的中风小鼠模型 研究并进一步了解cGKI-SMC信号在预防中风中的作用。这些 这些结果将为cGKI-Elevating化合物的开发提供科学基础 中风患者的治疗药物。