Transglutaminase 2 S-Nitrosylation: Role in Age-Related Vascular Stiffness

转谷氨酰胺酶 2 S-亚硝基化：在年龄相关血管僵硬中的作用

基本信息

批准号：
8502543
负责人：
DAN E BERKOWITZ
金额：
$ 37.52万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-20 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8502543
关键词：
Age Aging Animal Model Aorta Arteries Autopsy Biological Availability Blood Pressure Blood Vessels Buffers Cardiovascular Diseases Cardiovascular system Cell model Cell surface Characteristics Collagen Cystamine Cysteine Data Elastin Elderly Elements Endothelial Cells Endothelium Enzyme Inhibition Enzymes Equilibrium Exercise Extracellular Matrix Extracellular Matrix Proteins Fibroblasts Fracture Grant Heart Human Hypertension Inbred F344 Rats Isolated systolic hypertension Lead Location Measurement Measures Mediating Molecular Morbidity - disease rate Mus Myocardial Infarction NG-Nitroarginine Methyl Ester NOS3 gene Oxidation-Reduction Pathway interactions Perfusion Physiologic pulse Physiological Post-Translational Protein Processing Property Proteins Rattus Reactive Oxygen Species Regulation Risk Risk Factors Role SKIL gene Signal Transduction Silastic Smooth Muscle Myocytes Stroke System Tissues Transglutaminases Up-Regulation Vascular Endothelial Cell Vascular Endothelium Vascular Smooth Muscle age effect age related blood pump crosslink in vivo inhibitor/antagonist mortality pressure protein protein interaction public health relevance research study scaffold shear stress transcription factor transglutaminase 2

项目摘要

DESCRIPTION (provided by applicant): Vascular stiffness has been clearly established as a risk factor for cardiovascular disease, and is an independent predictor of cardiovascular morbidity and mortality. Aging is associated with increased vascular stiffness and isolated systolic hypertension, which results from alterations in the properties of all elements of the vascular wall including endothelium, vascular smooth muscle, and matrix. Although both dynamic (alterations in endothelial function and effects on vascular smooth contractility), as well as structural (eg. fracturing of elastin) have been described in aging, molecular mechanisms underlying age-related vascular stiffness remain poorly understood. Thus targeted therapy remains elusive. Tissue transglutaminase (TG2, tTG), expressed in vascular endothelial, smooth muscle cells, and fibroblasts, enzymatically form cross-links between extracellular matrix proteins, and may contribute to this pathobiology. Ca2+dependent activation of TG2 is dependent on its externalization to the cell surface. Recently, it has been demonstrated that S- nitrosylation, a redox-sensitive post-translational modification of cysteine residues, leads to TG2 enzyme inhibition. In Preliminary Data, we demonstrate that TG2 is S-nitrosylated in cellular models and that this leads to decreased cell-surface localization and decreased cross-linking activity. Furthermore, using TG2-/- mice, we show that TG2 is the primary TGase mediating stiffness of conduit arteries, and that it is regulated by endothelium-derived NO. We further demonstrate that in aging rat and human aorta, TG2 activity is increased, and it's S-nitrosylation is decreased despite unchanged TG2 abundance. Finally, inhibition of TG in old rats reduces vascular stiffness. It is now well established that NO bioavailability is diminished and reactive oxygen species (ROS) are increased in aging. Given this change in the nitroso-redox balance, we hypothesize that aging is associated with decreased TG2 S-nitrosylation and therefore, increased externalization and increased matrix cross-linking activity and TG2-dependent downstream signaling. Together, these result in increased vessel stiffness, and ultimately impaired vascular function, a hallmark of aging. In this grant, we propose to determine the role of NO in the regulation of TG2 location, activity and downstream signaling, and determine whether this enzyme is a critical target in age-related vascular stiffness. We will use a hierarchical approach including endothelial cells, vascular tissue from young and old Fischer 344 rats and TG2-/- and NOS3-/- mice, and invasive and non-invasive measures of vascular characteristics in these animal models. The following are the specific aims: 1) To determine the role of endothelium-dependent NO in the of regulation of TG2 subcellular distribution and activity. 2) To determine the role of TGF2 in downstream TG2 mediated vascular signaling. 3) To determine the role of TG2 and its regulation by NO in age-related vascular stiffness in animal models using sophisticated measures of vascular properties in vivo. 4) To determine TG2 activity in the aorta of aging humans.

描述（由申请人提供）：显然已将血管僵硬确定为心血管疾病的危险因素，并且是心血管发病率和死亡率的独立预测指标。衰老与血管刚度增加和分离的收缩期高血压有关，这是由于血管壁的所有元素（包括内皮，血管平滑肌和基质）的特性的改变而导致的。尽管在衰老中描述了动态（内皮功能的改变和对血管平滑收缩性的影响），以及结构性（例如弹性蛋白的破裂），但与年龄相关的血管僵硬的分子机制在衰老中都尚不清楚。因此，有针对性的治疗仍然难以捉摸。在血管内皮，平滑肌细胞和成纤维细胞中表达的组织转谷氨酰胺酶（TG2，TTG）在细胞外基质蛋白之间形成交联，可能有助于这种病理生物学。 Ca2+依赖性TG2的激活取决于其对细胞表面的外部化。最近，已经证明S-亚硝基化是对半胱氨酸残基的氧化还原敏感的翻译后修饰，导致TG2酶抑制作用。在初步数据中，我们证明了TG2在细胞模型中已被S-亚硝基化，这会导致细胞表面定位降低和交联活性降低。此外，使用TG2 - / - 小鼠，我们表明TG2是导管动脉刚度的主要TGASE，并且由内皮衍生的NO调节。我们进一步证明，在衰老的大鼠和人主动脉中，TG2活性增加，尽管TG2丰度不变，但S-亚硝基化却降低了。最后，在老鼠中抑制TG可降低血管僵硬。现在已经很好地确定，衰老中没有生物利用度降低，活性氧（ROS）增加。鉴于硝基 - 雷克斯平衡的这种变化，我们假设衰老与TG2 S-硝基化的降低有关，因此，外部化增加和矩阵交联活性增加和TG2依赖性下游信号传导。共同，这些导致血管刚度的增加，并最终损害了血管功能，这是衰老的标志。在这笔赠款中，我们建议确定NO在TG2位置，活动和下游信号传导调节中的作用，并确定该酶是否是与年龄相关的血管刚度的关键靶标。我们将使用一种分层方法，包括内皮细胞，来自年轻和老年Fischer 344大鼠的血管组织，TG2 - / - 以及NOS3 - / - 小鼠，以及这些动物模型中血管特征的侵入性和非侵入性测量。以下是具体目的：1）确定内皮依赖性NO在TG2亚细胞分布和活性调节中的作用。 2）确定TGF2在下游TG2介导的血管信号传导中的作用。 3）使用体内的血管特性的精致测量方法，在动物模型中，在与年龄相关的血管僵硬中确定TG2及其调节的作用。 4）确定衰老人类主动脉的TG2活性。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

About fACE: perioperative use of angiotensin-converting enzyme inhibitors.

关于 fACE：围手术期使用血管紧张素转换酶抑制剂。

DOI：
10.1161/circulationaha.112.117671
发表时间：
2012
期刊：
Circulation
影响因子：
37.8
作者：
Barodka,Viachaslau;Nyhan,Daniel;Berkowitz,Dan
通讯作者：
Berkowitz,Dan

MPST but not CSE is the primary regulator of hydrogen sulfide production and function in the coronary artery.

DOI：
10.1152/ajpheart.00574.2014
发表时间：
2016-01-01
期刊：
American journal of physiology. Heart and circulatory physiology
影响因子：
0
作者：
Kuo, Maggie M;Kim, Dae Hee;Santhanam, Lakshmi
通讯作者：
Santhanam, Lakshmi

Tissue Transglutaminase Modulates Vascular Stiffness and Function Through Crosslinking-Dependent and Crosslinking-Independent Functions.

DOI：
10.1161/jaha.116.004161
发表时间：
2017-02-03
期刊：
Journal of the American Heart Association
影响因子：
5.4
作者：
Steppan J;Bergman Y;Viegas K;Armstrong D;Tan S;Wang H;Melucci S;Hori D;Park SY;Barreto SF;Isak A;Jandu S;Flavahan N;Butlin M;An SS;Avolio A;Berkowitz DE;Halushka MK;Santhanam L
通讯作者：
Santhanam L

Vasopressin-mediated enhancement of adrenergic vasoconstriction involves both the tyrosine kinase and the protein kinase C pathways.

加压素介导的肾上腺素能血管收缩增强涉及酪氨酸激酶和蛋白激酶 C 途径。

DOI：
10.1213/ane.0b013e3182691c11
发表时间：
2012
期刊：
Anesthesia and analgesia
影响因子：
5.7
作者：
Steppan,Jochen;Nyhan,SineádM;Sikka,Gautam;Uribe,Jorge;Ahuja,Ayushi;White,AnthonyR;Shoukas,ArtinA;Berkowitz,DanE
通讯作者：
Berkowitz,DanE

Nitric oxide regulates non-classical secretion of tissue transglutaminase.

一氧化氮调节组织转谷氨酰胺酶的非经典分泌。