Anaplerotic reprogramming of endothelial cells in pulmonary hypertension.

肺动脉高压中内皮细胞的回补重编程。

基本信息

批准号：
9323563
负责人：
Ruslan Rafikov
金额：
$ 38.38万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9323563
关键词：
3-nitrotyrosine AKT inhibition Affect Affinity Antioxidants Apoptosis Apoptotic Attenuated Binding Blood Vessels Cell Proliferation Cell Survival Cell physiology Cellular Stress Cessation of life Characteristics Charge Citric Acid Cycle Data Disease Endothelial Cells Ensure Equilibrium Event FRAP1 gene Heart failure Hypoxia Inhibition of Apoptosis Inhibition of Cell Proliferation Knockout Mice Lesion Lung Malignant Neoplasms Mediating Mitochondria Modeling Modification Nitrates Oxaloacetates Oxidative Phosphorylation Oxidative Stress PDPK1 gene PIK3CG gene PTEN gene Pathologic Pathway interactions Patients Peptides Peroxonitrite Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Play Production Proteins Publishing Pulmonary Hypertension Pulmonary artery structure Pyruvate Pyruvate Carboxylase Rattus Reaction Resistance Role SU 5416 Serine Signal Transduction Stress Response Signaling Structure Structure of parenchyma of lung Superoxides Testing Therapeutic Tyrosine Vascular remodeling Work biological adaptation to stress cell growth endothelial dysfunction lung hypoxia mitochondrial dysfunction nitration novel pressure pulmonary arterial hypertension

项目摘要

PROJECT SUMMARY Pulmonary arterial hypertension (PH) is a fatal disease with uncontrolled pulmonary vascular cell proliferation. Excessive endothelial cell (EC) growth leads to the formation of plexiform lesions, which are a characteristic cancer-like change within pulmonary arteries of patients with PH. The increased oxidative stress in lungs of patients with PH leads to the reaction of superoxide with NO. This results in the formation of highly reactive peroxynitrite (ONOO-) which, in turn, can produce nitro-tyrosine modifications in proteins. Lung tissues from patients with PH have an increased level of protein nitration. Our recently published work indicates that the nitration of tyrosine 350 residue in Akt leads to the sustained activation of Akt signaling. The activation of Akt is a well described cell survival and stress response but Akt activation is tightly regulated by the regulatory kinases/phosphatases (PTEN, PI3K, PDK1 and mTOR) that control Akt activity and ensuring that it is only temporarily active. Pathological activation of the Akt pathway has been implicated in various cancers. We hypothesize that in PH, nitration of Akt increases translocation of eNOS to mitochondria, which downregulates oxidative phosphorylation and activates Pyruvate Carboxylase (PC), which upregulates anaplerosis. This activation of anaplerosis then leads to pathological hyper-proliferation of endothelial cells (EC) and plexiform lesion formation in PH. Further, we propose to target pathological Akt signaling using an Akt- binding peptide conjugated with an antioxidant (shielding peptide) that only affects nitration mediated Akt activation. We will test these hypotheses in the following Aims: AIM 1: To elucidate the role of nitration (Y350) mediated Akt activation in activation of anaplerosis in EC. AIM 2: To determine whether inhibition of Akt nitration disrupts formation of the apoptosis resistant and proliferative EC phenotype. AIM 3: To examine the effect of Akt shielding peptide on EC hyper-proliferation in the SU5416/hypoxia PH models.

项目摘要肺动脉高压（pH）是一种致命疾病，具有不受控制的肺血管细胞增殖。过多的内皮细胞（EC）生长导致形成丛状病变，这是 pH患者的肺动脉动脉中的特征性癌症变化。增加 pH患者肺中的氧化应激导致超氧化物与NO的反应。这导致高度反应性过氧亚硝酸盐（ONOO-）的形成又可以产生硝基酪氨酸蛋白质的修饰。来自pH患者的肺组织的蛋白质硝化水平增加。我们最近发表的工作表明，Akt酪氨酸350残留物的硝化导致 AKT信号的持续激活。 AKT的激活是描述的细胞存活和应力反应但AKT激活受调节激酶/磷酸酶（PTEN，PI3K，，PI3K， PDK1和MTOR）控制AKT活动并确保其仅暂时活跃。病理 AKT途径的激活与各种癌症有关。我们假设在pH中， Akt的硝化增加了eNOS易位到线粒体，从而下调氧化磷酸化并激活丙酮酸羧化酶（PC），该羧化酶上调了无菌病。这然后，无链球菌的激活导致内皮细胞（EC）和 pH中的丛状病变形成。此外，我们建议使用Akt-靶向病理AKT信号传导与抗氧化剂（屏蔽肽）结合的结合肽，仅影响硝酸化介导的Akt 激活。我们将在以下目的中检验这些假设：目的1：阐明硝化作用（Y350）在EC中介导的AKT激活中的AKT激活。目标2：确定是否抑制 Akt硝化破坏了抗细胞凋亡的形成和增生性EC表型。目标3：到检查AKT屏蔽肽对SU5416/缺氧pH模型中EC高增殖的影响。