miRNA-regulation at focal adhesions establishes vascular mechanohomeostasis

粘着斑处的 miRNA 调节建立血管机械稳态

基本信息

批准号：
10656557
负责人：
Stefania Nicoli
金额：
$ 20.83万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10656557
关键词：
3&apos Untranslated Regions Actins Actomyosin Address Adhesions Aging Aorta Arteriosclerosis Atherosclerosis Basic Science Binding Biological Assay Blood Vessels Blood flow Buffers CRISPR/Cas technology Cardiovascular Diseases Cell Adhesion Cell Communication Cell Culture Techniques Cells Cerebrovascular system Cytoskeletal Modeling Cytoskeleton Data Development Disease Embryo Endothelial Cells Endothelium Environment Etiology Event Extracellular Matrix Focal Adhesions Gene Expression Gene Expression Regulation Genes Grant Hematopoietic Homeostasis Human Hypertension In Vitro Integrins Mechanical Stress Mechanics Mediating Medicine Messenger RNA Metabolism MicroRNAs Modeling Molecular Mutation Pathology Phenotype Physiological Poison Poisoning Post-Transcriptional Regulation Predisposition Process Property Proteins RNA Regulation Reporting Research Role Science Sentinel Stress Testing Time Tissues Transgenic Organisms Translating Vascular System Visualization Zebrafish blood vessel development cardiovascular health cell behavior hemodynamics in vivo insight mechanical behavior mechanical force mechanical properties mechanical signal mechanotransduction mutant nerve stem cell novel posttranscriptional protein complex resilience response spatiotemporal stem cell differentiation

项目摘要

PROJECT SUMMARY Endothelial cells are exceptional sentinels of tissues' mechanical properties. Endothelial cells sense the extracellular matrix of the vascular wall via integrin-based adhesions (focal adhesions), and homotypic adhesion between neighboring tissue cells to regulate changes in matrix composition, turnover and stiffening. Furthermore, endothelial cells sense and adjust to blood flow hemodynamics through dynamic processes involving changes in actomyosin and other cytoskeletal stresses, remodeling of focal adhesions and cell adhesions, and cytoskeleton reorganization. How endothelial cells can buffer changes in forces associated with both blood flow and extracellular matrix remains unclear. Here, we will study how endothelial cells regulate matrix and blood flow forces at focal adhesion, and how these mechanism(s) sustain vascular mechanical homeostasis. Cell homeostasis hinges on the activity of microRNAs (miRNAs) through a mechanism that is resilient and regulates gene expression rapidly. We recently discovered a miRNA network activated by changes in matrix stiffness. These miRNAs preferentially target cytoskeletal, adhesion and matrix (CAM) genes for post- transcriptional regulation of protein levels both in vitro and in vivo. We concluded that miRNA-dependent regulation of CAM genes is critical for mediating cell-matrix interactions and allows endothelial cells to maintain mechanical homeostasis under conditions of changing stiffness. Preliminary data presented in this application shows the novel and exciting observation that miRNAs and CAM mRNAs are uniquely localized at focal adhesions in endothelial cells. We hypothesize that CAM mRNAs are localized to and translated at focal adhesions, and that miRNAs post-transcriptionally regulate CAM protein levels at focal adhesions to buffer the effect of matrix and/or hemodynamic changes and maintain mechanical homeostasis. We will test this new hypothesis by setting up 2D cell culture models and the zebrafish vascular system to characterize matrix and/or hemodynamic properties of endothelial cells that lack of miRNA regulation at focal adhesions.

项目摘要内皮细胞是组织机械性能的特殊哨兵。内皮细胞感觉血管壁的细胞外基质通过基于整合素的粘附（焦点粘连）和同型相邻组织细胞之间的粘附以调节基质组成，周转和僵硬的变化。此外，内皮细胞感知并通过动态过程适应血流血流动力学涉及肌动球蛋白和其他细胞骨架应激的变化，重塑局灶性粘连和细胞粘连和细胞骨架重组。内皮细胞如何缓解相关力的力变化血流和细胞外基质都尚不清楚。在这里，我们将研究内皮细胞如何调节基质和局灶性粘附时的血液流动力，以及这些机制如何维持血管机械稳态。细胞稳态通过一种具有弹性的机制对microRNA（miRNA）的活性取决于并迅速调节基因表达。我们最近发现了一个通过变化激活的miRNA网络矩阵刚度。这些miRNA优先靶向细胞骨架，粘附和基质（CAM）基因体外和体内蛋白质水平的转录调节。我们得出结论，miRNA依赖性 CAM基因的调节对于介导细胞 - 矩阵相互作用至关重要，并允许内皮细胞维持在变化刚度的条件下，机械稳态。本申请中提供的初步数据显示了miRNA的新颖而令人兴奋的观察凸轮mRNA在内皮细胞中的局灶性粘连中独特地定位。我们假设凸轮 mRNA位于焦点粘附处和翻译，并在转录后进行miRNA 调节焦点粘附处的CAM蛋白水平，以缓冲基质和/或血液动力学的效果改变并保持机械稳态。我们将通过设置2D单元格来检验这一新假设培养模型和斑马鱼血管系统，以表征基质和/或血液动力学特性缺乏miRNA调节的内皮细胞在局灶性粘附处。