Mechanisms of Microvascular Remodeling Progression

微血管重塑进展机制

• 批准号: 9026447
• 负责人: Luis A Martinez-Lemus
• 金额: $ 37.11万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9026447
• 关键词: Actins; Animal Model; Animals; Arteries; Blood Vessels; Caliber; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cells; Clinical; Cytoskeleton; Data; Development; Disease Management; Ear; Elastin; Event; Excision; Extracellular Matrix; Gelatinase A; Goals; Hypertension; Individual; Intervention; Knowledge; LIM Domain Kinase 1; Lead; Life; MMP14 gene; Matrix Metalloproteinases; Measures; Modification; Molecular; Myocardial Infarction; Outcome; Patients; Peptides; Prevalence; Process; Production; Proteins; Public Health; Publications; Reporting; Research; Resistance; Resistance Process; Rho-associated kinase; Risk; Site; Smooth Muscle Myocytes; Staging; Stimulus; Stress Fibers; Stroke; Structure; Subcellular structure; Techniques; Technology; Testing; Therapeutic; Tissues; United States; Vascular Smooth Muscle; Vascular remodeling; Work; base; cofilin; constriction; crosslink; insight; intravital microscopy; normotensive; novel; novel strategies; polymerization; prevent; public health relevance; transglutaminase 2; vasoconstriction

 DESCRIPTION (provided by applicant): Vascular remodeling is an adaptive mechanism for long-term modification of vascular diameter. In hypertension, inward remodeling, that is, the structural reduction of the lumen diameter in resistance vessels, is associated with an increased risk for myocardial infarction and stroke. However, despite its prevalence and clinical importance, the mechanisms that control the inward remodeling process remain largely unknown. Our goal here is to identify mechanisms in the inward remodeling process of the resistance vasculature that may be intervened with novel strategies to prevent, stop or reverse the remodeling process, and consequently diminish the life-threatening cardiovascular events associated with it. Current publications and our own preliminary data indicate that tissue-type transglutaminase (TG2), LIM kinase (LIMK), and matrix metalloproteinase-2 (MMP2) within vascular smooth muscle cells (VSMC) are involved in the remodeling process. Therefore, as we and others have determined that inwardly remodeled resistance vessels have actin cytoskeletal structures that reduce their passive diameters and extracellular matrix (ECM) features characterized by a reduction in the number and size of fenestrae in the internal elastic lamina (IEL): Our hypothesis is that during the early stages of the inward remodeling process in resistance vessels, prolonged vasoconstriction leads to formation of permanent VSMC cytoskeletal structures via the intracellular activity of TG2 and LIM kinase, which in turn stimulate the production of MMP2 and the modification of the ECM, in particular the IEL. We will test our hypothesis in VSMC, isolated resistance arteries and a whole animal model of hypertension. Cells and tissues will come from animals, as well as from normotensive and hypertensive individuals. The expression and activity of the remodeling components tested in our hypotheses will be modulated using pharmacological and molecular means. Experimental outcomes will be measured using traditional and leading-edge techniques in protein and enzymatic activity analyses, as well as, atomic force, multiphoton, and long-term intravital microscopy. Our specific aims will test the hypotheses that: 1) Intracellular TG2 activates RhoA, Rho kinase and LIMK to phosphorylate and inactivate cofilin to favor formation of actin networks and stress-fibers, with TG2 further crosslinking actin structures to make them more persistent; and 2) that LIMK activates MMP14 and leads to expression/secretion of MMP2 from VSMC. Then MMP2 through its elastolytic actions generates elastin peptides that activate VSMC to produce more elastin. This new elastin is incorporated in the IEL and reduces the size and number of fenestrae in the IEL. We expect this study will provide new insights on how cytoskeletal and IEL structures of resistance arteries are modified in hypertension. This knowledge should have a positive impact on strategies for preventing and treating hypertension, and the management of diseases associated with vascular remodeling.

 描述（由申请人提供）：血管重塑是一种长期改变血管直径的适应性机制。在高血压中，向内重构，即阻力血管中管腔直径的结构性减小，与心肌梗死和中风的风险增加相关。然而，尽管它的流行和临床重要性，控制向内重塑过程的机制在很大程度上仍然未知。我们的目标是确定阻力血管系统向内重塑过程中的机制，这些机制可以通过新的策略进行干预，以预防、停止或逆转重塑过程，从而减少与之相关的危及生命的心血管事件。血管平滑肌细胞（VSMC）内的基质金属蛋白酶2（MMP 2）参与重塑过程。因此，正如我们和其他人已经确定的那样，向内重塑的阻力血管具有肌动蛋白细胞骨架结构，其减少了它们的被动直径和细胞外基质（ECM）特征，其特征在于内弹性膜（IEL）中窗孔的数量和大小减少：我们的假设是，在阻力血管向内重塑过程的早期阶段，延长的血管收缩通过TG 2和LIM激酶的细胞内活性导致永久的VSMC细胞骨架结构的形成，其又刺激MMP 2的产生和ECM，特别是IEL的修饰。 我们将在VSMC、分离的阻力动脉和整个高血压动物模型中检验我们的假设。细胞和组织将来自动物，以及来自血压正常和高血压个体。在我们的假设中测试的重塑组分的表达和活性将使用药理学和分子手段来调节。实验结果将使用蛋白质和酶活性分析中的传统和前沿技术，以及原子力，多光子和长期活体显微镜进行测量。我们的具体目标将测试以下假设：1）细胞内TG 2激活RhoA、Rho激酶和LIMK以磷酸化和磷酸化微丝蛋白以促进肌动蛋白网络和应力纤维的形成，其中TG 2进一步交联肌动蛋白结构以使其更持久;和2）LIMK激活MMP 14并导致MMP 2从VSMC表达/分泌。然后MMP 2通过其弹性蛋白溶解作用产生弹性蛋白肽，激活VSMC产生更多的弹性蛋白。这种新的弹性蛋白被纳入IEL中，并减少IEL中窗孔的大小和数量。 我们希望这项研究将提供新的见解，细胞骨架和IEL结构的阻力动脉是如何修改高血压。这些知识应该对预防和治疗高血压的策略以及与血管重塑相关的疾病的管理产生积极的影响。