Lysyl Oxidase Like 2: A Novel Target in Aging Associated Vascular Stiffening

赖氨酰氧化酶样 2：衰老相关血管硬化的新靶点

• 批准号: 10837659
• 负责人: Lakshmi Santhanam
• 金额: $ 11.14万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10837659
• 关键词: Age; Aging; Animal Model; Architecture; Behavior; Biochemical; Biochemistry; Biological Availability; Biology; Blood Vessels; Cardiovascular system; Caring; Cause of Death; Cells; Cellular Assay; Clinical; Clinical Management; Collagen; Cytoskeleton; Data; Deposition; Deterioration; Development; Disease; Elderly; Environment; Essential Hypertension; Etiology; Event; Extracellular Matrix; Extracellular Matrix Proteins; Factor Xa; Fibrosis; Financial Hardship; Foundations; Functional disorder; Genetic; Goals; Grant; Hypertension; In Vitro; Interdisciplinary Study; Interruption; Investigation; Knock-out; Knowledge; LOX gene; LOXL2 gene; Laboratories; Lead; Life Style Modification; Measures; Mechanics; Mediating; Mission; Molecular; Molecular Biology; Morbidity - disease rate; Mus; Myography; National Heart, Lung, and Blood Institute; Nitric Oxide; Physiologic pulse; Population; Pre-Clinical Model; Protein Biochemistry; Proteins; Proteolytic Processing; Proteomics; Public Health; Reagent; Regulation; Role; SRCR proteins; Scaffolding Protein; Scavenger Receptor Cysteine-Rich Domain; Scheme; Smooth Muscle Myocytes; Source; Substrate Specificity; Syncope; Tamoxifen; Testing; Therapeutic; Translating; Vascular Smooth Muscle; Work; Youth; age related; aging population; amine oxidase; crosslink; demographics; effective therapy; experimental study; falls; in vivo; innovation; insight; middle age; mortality; mouse model; multidisciplinary; novel; novel therapeutics; pharmacologic; pressure; prevent; protein protein interaction; scaffold; targeted agent; targeted treatment

Project summary: Increased central vascular stiffness in aging is an independent predictor of major adverse cardiovascular events and the leading cause of morbidity and mortality in the elderly. While mechanisms of vascular stiffening are emerging, targeted therapies are not yet available. Vascular stiffening and its sequelae remain clinically challenging to manage and currently, care relies on lifestyle modifications and pharmacologic agents targeting essential hypertension despite the distinct etiology and cellular molecular mechanisms, leading to syncope and falls in the elderly. and with the shift of population demographics towards the elderly, the clinical, societal, and financial burden of vascular stiffening has grown. Therefore, there is an urgent need to identify and characterize potential targets for clinical management of vascular stiffening and mitigate its sequelae. Using an unbiased proteomic screen, we have identified lysyl oxidase like 2 (LOXL2) as a novel protein that mediates vascular stiffening. Our hypothesis, strongly supported by preliminary data, is that LOXL2 regulates matrix composition, deposition, and architecture, modulates vascular smooth muscle cell (VSMC) micromechanics and contractility, and contributes to aging associated increase in vascular stiffness in mice. In this grant, we propose to examine the mechanisms of LOXL2-mediated vascular stiffening in aging in a hierarchical fashion from cells to mouse models of aging. We have generated novel reagents, mouse models, and assembled a strong, multidisciplinary research team to successfully execute the proposed work. In Aim 1, we will determine the mechanism of LOXL2 regulation by nitric oxide and proteolytic processing by Factor Xa in VSMCs. These aspects of LOXL2 regulation in cells and vasculature are not yet known. In Aim 2, we postulate that the scavenger receptor cysteine rich (SRCR) domains and amine oxidase (LOX) domains present in LOXL2 have distinct functions that contribute uniquely to vascular stiffening. We will 1) define the mechanisms of ECM remodeling by LOXL2 with specific emphasis on its putative scaffolding function and 2) determine how LOXL2 modulates VSMC micromechanics, cytoskeletal dynamics, and behavior/function. In Aim 3, we will determine if targeting LOXL2 mitigates age-associated vascular stiffening in mouse models. Using a combination of genetic (tamoxifen inducible global, VSMC-specific, and EC-specific LOXL2-/- mice), and pharmacologic (PAT-1251) approaches to target LOXL2, we will identify 1) the cellular source of LOXL2 in the aging vessel, 2) investigate if LOXL2 targeting can halt or reverse vascular stiffening in aging, and 3) identify whether VSMC stiffening or ECM deposition is the primary mechanism by which LOXL2 acts in the aging vessel. In sum, these studies will advance our knowledge of LOXL2 protein biochemistry and biology, establish the mechanisms by which LOXL2 is regulated in the vasculature, determine the mechanisms by which LOXL2 drives age-associated vascular stiffening, and lay the scientific foundation for the discovery and development of novel and effective therapies for the clinical management of vascular stiffening.

项目总结： 衰老时中心血管僵硬增加是主要不良心血管疾病的独立预测因子 这些事件是导致老年人发病和死亡的主要原因。而血管僵硬的机制 都是新兴的，靶向治疗还没有可用的。血管硬化及其后遗症仍在临床上存在 管理具有挑战性，目前，护理依赖于生活方式的改变和药物靶向 尽管有不同的病因和细胞分子机制，但原发性高血压会导致晕厥和 跌倒在老年人身上。随着人口结构向老年人的转变，临床、社会和 血管硬化带来的财政负担增加了。因此，迫切需要确定和执行 确定血管硬化临床治疗的潜在靶点，并减轻其后遗症。 利用蛋白质组学无偏筛选，我们鉴定了赖氨酰氧化酶样蛋白2(LOXL2)是一种新的蛋白质 这会导致血管僵硬。我们的假设得到了初步数据的有力支持，即LOXL2 调节基质组成、沉积和结构，调节血管平滑肌细胞(VSMC) 微观力学和收缩能力，并有助于与衰老相关的小鼠血管僵硬的增加。在……里面 这项授权，我们建议研究LOXL2介导的血管硬化在衰老中的机制。 层次分明的方式从细胞老化到老鼠模型。我们已经产生了新的试剂，老鼠模型， 并组建了一支强大的、多学科的研究团队，成功地执行了拟议的工作。 在目标1中，我们将确定一氧化氮和蛋白质降解过程对LOXL2的调节机制 按VSMCs中的Xa因子计算。LOXL2在细胞和血管系统中调节的这些方面尚不清楚。在AIM 2、我们推测清道夫受体富含半胱氨酸(SRCR)结构域和胺氧化酶(LOX)结构域 存在于LOXL2中，具有独特的促进血管硬化的独特功能。我们将1)定义 LOXL2重塑ECM的机制及其可能的支架作用2) 确定LOXL2如何调节VSMC的微观力学、细胞骨架动力学和行为/功能。在……里面 目的3，我们将确定靶向LOXL2是否能减轻小鼠模型中与年龄相关的血管僵硬。 使用遗传(他莫昔芬可诱导的全球、VSMC特异性和EC特异性LOXL2-/-小鼠)的组合， 和药理学(PAT-1251)靶向LOXL2的方法，我们将确定1)LOXL2的细胞来源 老化的血管，2)研究LOXL2靶向是否可以阻止或逆转衰老中的血管硬化，以及3) 确定VSMC硬化或ECM沉积是LOXL2在 老旧的船只。总之，这些研究将促进我们对LOXL2蛋白生物化学和生物学的了解， 建立LOXL2在血管系统中的调节机制，通过 其中LOXL2驱动与年龄相关的血管硬化，并为发现和 开发新的有效的治疗方法用于临床治疗血管僵硬。

项目成果

Lysyl oxidase-like 2 processing by factor Xa modulates its activity and substrate preference.

• DOI: 10.1038/s42003-023-04748-8
• 发表时间: 2023-04-07
• 期刊: COMMUNICATIONS BIOLOGY
• 影响因子: 5.9
• 作者: Wang, Huilei;Poe, Alan;Martinez Yus, Marta;Pak, Lydia;Nandakumar, Kavitha;Santhanam, Lakshmi
• 通讯作者: Santhanam, Lakshmi

Mitochondrial-triggered immune responses mechanistically connect drug-induced steatohepatitis and cardiomyopathy associated with nonalcoholic steatohepatitis.

线粒体触发的免疫反应在机制上将药物引起的脂肪性肝炎和与非酒精性脂肪性肝炎相关的心肌病联系起来。

• DOI: 10.1038/s41423-021-00724-6
• 发表时间: 2021
• 期刊: Cellular & molecular immunology
• 影响因子: 24.1
• 作者: Wang,Haoran;Pai,Sananda;Ishiyama,Shun;Guo,Xin;Gao,WeiDong;Santhanam,Lakshmi;Gabrielson,Kathy;Njoku,DoloresB
• 通讯作者: Njoku,DoloresB