Tension-induced SGK-1 Signaling in AAA

AAA 中张力诱导的 SGK-1 信号转导

• 批准号: 10368068
• 负责人: Jean Marie Ruddy
• 金额: $ 17.65万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-14 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10368068
• 关键词: Abdominal Aortic Aneurysm; Affect; American; Aneurysm; Angiotensin II; Aorta; Aortic Aneurysm; Atherosclerosis; Attenuated; Automobile Driving; Biochemical; Biomechanics; Blood Pressure; Bone Marrow; CCL2 gene; Caliber; Cardiovascular system; Cell Culture Techniques; Cell Line; Cell Membrane Permeability; Cells; Critical Thinking; Culture Media; Dependence; Development; Disease; Enzyme-Linked Immunosorbent Assay; Epidemiology; Experimental Designs; Exposure to; Flow Cytometry; Future; Gelatinase B; Glucocorticoids; Growth; Harvest; Hyperplasia; Hypertension; ITGAM gene; Immunoblot Analysis; In Vitro; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Infusion procedures; Innovative Therapy; Interleukin-6; Knock-out; Knockout Mice; Laboratories; Laboratory Research; Life; Link; Measures; Mechanics; Mediating; Mentorship; Microscopy; Modeling; Molecular; Mouse Strains; Mus; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Peptides; Pharmacotherapy; Phosphotransferases; Plasma; Play; Population; Procedures; Production; Proteins; Pulmonary Hypertension; Regulation; Research Methodology; Research Personnel; Resources; Role; Rupture; Serum; Signal Transduction; Small Interfering RNA; Smooth Muscle Myocytes; Stains; Stimulus; Stretching; Stroke; Tissues; Unstable angina; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; Wild Type Mouse; Work; abdominal aorta; blood pressure elevation; cardiovascular risk factor; career; cytokine; digital; hypertensive; in vivo; inhibitor; inhibitor therapy; macrophage; migration; monocyte; mortality; mortality risk; mouse model; new therapeutic target; response; skills; transcription factor; vascular inflammation

Hypertension (HTN) is a major risk factor for cardiovascular mortality and greater than 30% of Americans are currently affected. The impact of elevated blood pressure on aortic wall remodeling has significant implications for vascular disease in general, and aneurysm formation specifically. AngiotensinII (AngII) is a potent vasoactive peptide and contributes to vascular inflammation by stimulating production of interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) to augment the population of macrophages within the aortic wall. Interestingly, patients with abdominal aortic aneurysms (AAA) have also consistently demonstrated elevated plasma and aortic tissue levels of IL-6, along with dense macrophage infiltration. Defining the tension-induced kinase driving cytokine production is vital to identifying a potential target for pharmacotherapy of AAA. Recent evidence has identified serum and glucocorticoid inducible kinase-1 (SGK-1) as a mechanically sensitive kinase that contributes to intimal hyperplasia, atherosclerosis, and pulmonary hypertension, and its downstream regulation of several transcription factors can modulate the production of IL-6 and MCP-1. Therefore, it is hypothesized that HTN promotes VSMC activation of SGK-1 to produce pro-inflammatory cytokines that accumulate macrophages to propagate AAA growth. The first specific aim will analyze tension-induced SGK-1 activation and cytokine expression from aortic VSMCs harvested from C57Bl/6 wild-type and SGK-1 knockout (SGK-1KO) mice. The ability of this conditioned media to stimulate monocyte/macrophage migration through a permeable membrane will also be assessed. In the second aim, C57Bl/6 and SGK-1KO mice with induced HTN (via AngII infusion) will be evaluated for the activation of SGK-1, production of pro-inflammatory cytokines IL-6 and MCP-1, and accumulation of macrophages. Dependence of this inflammatory response on SGK-1 activity will be further explored by treating wild-type mice subjected to induced HTN with the selective SGK-1 inhibitor EMD638683. The third specific aim will focus on the role of SGK-1 in initiating and propagating AAA formation by employing a validated model of AAA induction with CaCl2 application and treating mice with EMD638683. The terminal procedure will evaluate AAA diameter and production of target cytokines and proteases. The opportunity for elevated tension to augment this effect will be explored by inducing HTN and concurrent AAA in the two mouse strains and conducting parallel biochemical analysis. By establishing this link between HTN, inflammation, and the initiation of degradative vascular remodeling through the activity of SGK-1, future studies may utilize targeted inhibitor therapies to attenuate aneurysmal degeneration in the abdominal aorta. Moreover, the experimentation outlined in this project will provide an opportunity for Dr. Ruddy to engage the resources of MUSC and the Cardiovascular Research Laboratory to expand her research methodology, develop mentorship qualities, enhance her critical-thinking skills, and augment her grantsmanship to propel her toward a career as an independent researcher.

高血压（HTN）是心血管死亡率的主要危险因素，大于30％的美国人 目前受到影响。升高血压对主动脉壁重塑的影响显着 一般来说，对血管疾病的意义和动脉瘤形成。 Angiotensinii（Angii）是有效的 血管活性肽并通过刺激白介素6（IL-6）的产生和 单核细胞趋化剂蛋白1（MCP-1）增加主动脉壁内巨噬细胞的种群。 有趣的是，腹主动脉瘤（AAA）的患者也始终显示出升高 IL-6的血浆和主动脉组织水平以及密集的巨噬细胞浸润。定义张力引起的 激酶驱动细胞因子的产生对于确定AAA药物疗法的潜在靶标至关重要。最近的 证据已将血清和糖皮质激素诱导激酶-1（SGK-1）确定为机械敏感的激酶 这有助于内膜增生，动脉粥样硬化和肺动脉高压及其下游 调节几种转录因子可以调节IL-6和MCP-1的产生。因此，是 假设HTN促进了SGK-1的VSMC激活，以产生促炎性细胞因子 积累巨噬细胞以传播AAA的生长。第一个特定目的将分析张力引起的SGK-1 从C57BL/6野生型和SGK-1基因敲除从主动脉VSMC中激活和细胞因子表达 （SGK-1KO）小鼠。这种条件培养基刺激单核细胞/巨噬细胞迁移的能力 也将评估渗透膜。在第二个目标中，具有诱导HTN的C57BL/6和SGK-1KO小鼠 （通过Angii输注）将评估SGK-1的激活，产生促炎性细胞因子IL-6 和MCP-1，以及巨噬细胞的积累。这种炎症反应对SGK-1活性的依赖性 将通过选择性SGK-1抑制剂治疗诱导HTN的野生型小鼠进一步探索 EMD638683。第三个具体目标将重点介绍SGK-1在启动和传播AAA形成中的作用 通过使用CACL2应用使用经过验证的AAA诱导模型，并使用EMD638683处理小鼠。这 终端程序将评估靶细胞因子和蛋白酶的AAA直径和产生。机会 为了增加张力以增加这种效果，将通过在两者中诱导HTN和同时AAA来探索这种效果 小鼠菌株和进行平行的生化分析。通过在HTN之间建立这种联系， 炎症，以及通过SGK-1的活性，未来研究的降解血管重塑的开始 可以利用靶向抑制剂疗法来减轻腹主动脉中的动脉瘤变性。而且， 该项目中概述的实验将为Ruddy博士提供机会参与 MUSC和心血管研究实验室，以扩大她的研究方法，开发指导 素质，提高她的批判性技巧，并增强她的赠款技巧，以推动她的职业生涯 独立研究人员。