Progression of Arterial Aging: the Local MCP-1/MMP-2/TGF-beta 1 Signaling Loop

动脉老化的进展：局部 MCP-1/MMP-2/TGF-beta 1 信号环路

• 批准号: 10913025
• 负责人: Mingyi Wang
• 金额: $ 4.7万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10913025
• 关键词: Acceleration; Actins; Age; Age-associated memory impairment; Aging; Angiotensin II; Angiotensin II Receptor; Animals; Aorta; Apoptosis; Apoptotic; Atherosclerosis; Attenuated; Blood Pressure; Blood Vessels; CC chemokine receptor 2; CCL2 gene; Cardiac; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Cell Cycle; Cell Separation; Cells; Cellularity; Chest; Chronic; Collaborations; Collagen; Collagen Type I; Crossbreeding; Cytoskeleton; Deceleration; Dementia; Deposition; Development; Down-Regulation; ETS1 gene; Elastic Fiber; Elastin; Elderly; Elements; Equilibrium; Erythropoietin; Exhibits; Experimental Models; Extracellular Matrix; Fibroblasts; Fibronectins; Fibrosis; Friction; Gelatinases; Genes; Genetic Transcription; Heart; Heterogeneity; Hypertension; Impaired cognition; In Situ; In Vitro; Individual; Inflammation; Inflammatory; Integrins; Interstitial Collagenase; Invaded; Knockout Mice; Length; Leucocytic infiltrate; Life; Link; Losartan; MMP2 gene; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Mechanics; Mediating; Messenger RNA; Metalloproteins; MicroRNAs; Molecular; Mothers; Muscle Cells; Myocardial; Myocarditis; Myocardium; Organism; Phenotype; Phosphorylation; Physiological; Play; Predisposition; Production; Proliferating; Proteins; Rattus; Rattus norvegicus; Reactive Oxygen Species; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Signaling Molecule; Small Interfering RNA; Smooth Muscle Myocytes; Stroke; TGFB1 gene; Therapeutic; Time; Tissues; Translations; Tropoelastin; Up-Regulation; Vascular Smooth Muscle; Western Blotting; age related; aged; antagonist; cardioprotection; cytokine; discoidin domain receptor 2; frailty; healthspan; human model; in vivo; inhibitor; juvenile animal; knock-down; mechanical properties; monocyte; myocardial injury; nonhuman primate; novel strategies; preservation; pressure; prevent; proendothelin 1; protein aminoacid sequence; receptor; response; senescence; therapy design; vascular smooth muscle cell proliferation; wound healing

The coexistence of vascular smooth muscle cell (VSMC) cellularity via invasion and proliferation and excessive collagen deposition within a diffusely thickened intima is a hallmark of central arterial wall inflammatory remodeling that accompanies advancing age. However, the molecular and cellular mechanisms involved remain undefined. Immunostaining, immunoblotting, and RT-PCR of rat aortic walls demonstrate that a triad of proinflammatory molecules monocyte chemoattractant protein-1 (MCP-1), transforming growth factor beta 1 (TGF-1), and matrix metalloprotein type II (MMP-2) mRNA and protein increase within the aging aortic wall. Treating VSMCs, isolated from 8-mo-old Fisher 344 crossbreed Brown Norway (FBN) rats (young) with MCP-1, via the CC-chemokine receptor 2 (CCR-2), promotes both an increase in TGF-1 activity, up to levels of untreated VSMCs isolated from 30-mo-old (old) FXBN rats in culture, and a concurrent increase in MMP-2 activation. Furthermore, treating young VSMCs with TGF-1 increases the levels of MCP-1 and MMP-2 activation, to levels of untreated VSMC isolated from old rats. This autocatalytic signaling loop that enhances collagen production and invasiveness of VSMCs is effectively suppressed by silencing the MCP-1 gene, treating with a CCR2 antagonist, or through the inhibition of MMP-2. Certain levels of activated MCP-1, MMP-2, or TGF-1 trigger a feed-forward signaling mechanism that potentially drives the initiation and progression of age-associated adverse arterial senescent proinflammatory remodeling. The active MMP-associated activation of TGF-1 plays an important role in the stiffening of aging VSMCs and arterial walls. Distinct mechanical properties of primary VSMCs isolated from thoracic aortae of young vs. aged F344XBN rats have been observed. Individual VSMCs derived from aged animals shows an internal network of the actin cytoskeleton exhibiting increased stiffness and frictional (loss) moduli compared with those cells derived from the young animals. This discrete mechanical response is long-lived in culture and is persistent across a physiological range of matrix rigidity. Strikingly, TGF-1 has been reported to emerge as a specific modifier of age-associated VSMC stiffening. TGF-1 reinforces the mechanical phenotype of arterial aging in VSMCs on multiple time and length scales through clustering of mechanosensitive receptors 51 and v3 integrins. Notably, similar observations have been found in the perivascular adventitial fibroblasts with advancing age, The broad-based inhibition of TGF-1 expression or signal transduction in VSMCs and adventitial fibroblasts may be a useful therapeutic approach to mitigate the age associated arterial wall stiffening, hypertension, atherosclerosis, and age-associated vascular-related dementia. Inhibiting MMP-2 activation associated with TGF-1 activation decelerates age-associated arterial proinflammation and its attendant increase in arterial systolic blood pressure. Eight months of chronic administration of a broad-spectrum MMP inhibitor, PD166793, via a daily gavage, to 16-month-old rats markedly blunted the expected age-associated increases in arterial pressure. This was accompanied by the following effects: (1) inhibition of the age-associated increases in aortic gelatinase and interstitial collagenase activity in situ; (2) preservation of the elastic fiber network integrity; (3) a reduction of collagen deposition; (4) a reduction of MCP-1 and TGF-1 activation; (5) an increase of vasorin, an inhibitor of TGF-1 signaling; (6) a decrease in the activity of the profibrogenic signaling molecule SMAD 2/3 (Sma and Mad (Mothers against decapentaplegic)-2/3) phosphorylation; (7) an inhibition of proendothelin 1 activation; and (8)a downregulation of expression of V-Ets Avian Erythroblastosis Virus E26 Oncogene Homolog 1 (Ets-1). Importantly, MMP inhibition significantly reduce the degradation of tropoelastin in VSMCs, a core element of elastin fibrils. Collectively, MMP inhibition diminishes age-associated arterial proinflammation and age-associated increases in systolic blood pressure. A signaling interrelationship exists between angiotensin II (Ang II), TGF-1, MMP-2 and vasorin within aging VSMCs. In vivo studies in old (30-month-old) versus young FXBN rats have showed that the aortic transcription and translation levels of vasorin markedly decreases with aging. In vitro studies of early passage VSMCs from old versus young rat aortae indicate that the abundance of vasorin protein is substantially reduced. Ang II-associated reduction of vasorin protein abundance in young VSMCs and age-associated changes in vasorin protein levels are reversed when treated with Losartan, an Ang II receptor (AT1) antagonist, in both in vitro and in vivo conditions. In addition, age-associated arterial vasorin is closely associated with an enhanced capacity of MMP activation. Thus, maintaining the critical balance of the full-length vasorin/TGF-1 signaling is a novel approach to hinder age-associated MMP-2 activation and arterial adverse extracellular matrix remodeling. In collaboration with K Shivakumar, we have demonstrated that collagen type I-activated Discoidin Domain Receptor 2 (DDR2) mediates Ang II-stimulated transcriptional up-regulation of fibronectin by Yes-activated Protein in cardiac fibroblasts. Further, siRNA-mediated fibronectin knockdown attenuates Ang II-stimulated expression of collagen type I and anti-apoptotic cIAP2 and enhances susceptibility to apoptosis. Importantly, an obligate role for fibronectin has been observed in Ang II-stimulated expression of AT1R, the Ang II receptor, which potentially link extracellular matrix with Ang II signaling in cardiac fibroblasts. In vivo, modestly reduced basal levels of AT1R in DDR2-null mouse myocardium is associated with a reduction of myocardial Integrin-1 levels. The role of fibronectin, downstream of DDR2, could be a critical determinant of cardiac fibroblast-mediated wound healing following myocardial injury. Aging is associated with increased levels of reactive oxygen species and inflammation that disrupt cardiovascular function and leads to organism-wide frailty later in life. ARA290 (cibinetide), an 11-aa non-hematopoietic peptide sequence within the cardioprotective domain of erythropoietin, mediates tissue protection by reducing inflammation and fibrosis. ARA290 ameliorates age-associated cardiac inflammation and is linked to structural and functional changes in the heart. We found that chronic ARA290 treatment mitigated age-related increases in the cardiac non-myocyte to myocyte ratio, infiltrating leukocytes and monocytes, pro-inflammatory cytokines, total NF-B, and p-NF-B. Thus, administration of ARA290 reduces cell and tissue inflammation, mitigates structural and functional changes within the cardiovascular system with aging leading to amelioration of frailty and preserved health span. We also recently found that an increased microRNA 34 (miR34) in the arterial wall and VSMCs is known to accompany aging in rats and nonhuman primates, which strongly activate MMP2. Importantly, the age-associated increase in miR-34a was causally linked to Ang II signaling within arterial all or VSMCs. Conversely the age-associated increase in Ang II signaling was potentially associated with the miR-34a associated downregulation in Ang II Receptor-Associated Protein (AGTRAP) and situin1 (SIRT1) within the arterial walls or cells. In vitro studies show that MFG-E8 increases the proliferation of VSMCs via an acceleration of cell cycle while miR-34a increases the senescence and invasion of VSMCs via a deceleration of cell cycle and activation of MMP-2. Thus, controlling the age-associated arterial miR-34a is a potential approach to curbing Ang II-linked proinflammation and VSMC heterogeneity and slowing arterial aging

在弥漫性增厚的内膜内，血管平滑肌细胞（VSMC）的浸润和增殖以及过多的胶原沉积共存，是随着年龄增长而发生的中央动脉壁炎症性重构的标志。然而，所涉及的分子和细胞机制仍未明确。大鼠主动脉壁的免疫染色、免疫印迹和RT-PCR显示，促炎分子单核细胞化学吸引蛋白-1 （MCP-1）、转化生长因子-1 （TGF-1）和基质金属蛋白II (MMP-2) mRNA和蛋白在老化的主动脉壁内增加。用MCP-1通过cc趋化因子受体2 （CCR-2）处理从8mo龄的Fisher 344杂交褐挪威（FBN）大鼠（年轻）中分离的VSMCs，可促进TGF-1活性的增加，达到从30mo龄（老）FXBN大鼠中分离的未处理VSMCs的水平，并同时增加MMP-2的激活。此外，用TGF-1治疗年轻VSMC可使MCP-1和MMP-2的活化水平提高到未治疗的老年大鼠VSMC水平。通过沉默MCP-1基因、用CCR2拮抗剂治疗或抑制MMP-2，可以有效抑制这种增强胶原生成和VSMCs侵袭性的自催化信号环。一定水平的活化MCP-1、MMP-2或TGF-1触发前馈信号机制，潜在地驱动与年龄相关的不良动脉衰老促炎重塑的开始和进展。

项目成果

A small erythropoietin derived non-hematopoietic peptide reduces cardiac inflammation, attenuates age associated declines in heart function and prolongs healthspan.

• DOI: 10.3389/fcvm.2022.1096887
• 发表时间: 2022
• 期刊: Frontiers in cardiovascular medicine
• 影响因子: 3.6

Serum from calorie-restricted animals delays senescence and extends the lifespan of normal human fibroblasts in vitro.

卡路里限制动物的血清延迟衰老，并在体外延伸正常人成纤维细胞的寿命。

• DOI: 10.18632/aging.100719
• 发表时间: 2015-03
• 期刊: Aging
• 作者: de Cabo R;Liu L;Ali A;Price N;Zhang J;Wang M;Lakatta E;Irusta PM
• 通讯作者: Irusta PM

So! What's aging? Is cardiovascular aging a disease?

• DOI: 10.1016/j.yjmcc.2015.04.005
• 发表时间: 2015-06
• 期刊: Journal of molecular and cellular cardiology
• 影响因子: 5
• 作者: Lakatta EG

A local proinflammatory signalling loop facilitates adverse age-associated arterial remodeling.

• DOI: 10.1371/journal.pone.0016653
• 发表时间: 2011-02-08
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Wang M;Spinetti G;Monticone RE;Zhang J;Wu J;Jiang L;Khazan B;Telljohann R;Lakatta EG
• 通讯作者: Lakatta EG