Effect of shear stress on coronary smooth muscle maturation

剪切应力对冠状动脉平滑肌成熟的影响

• 批准号: 10580556
• 负责人: Laura Ann Dyer
• 金额: $ 39.42万
• 依托单位: UNIVERSITY OF PORTLAND
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-15 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580556
• 关键词: Address; Adult; Aorta; Arteries; Attention; Blood; Blood flow; Chick Embryo; Coronary; Coronary Vessel Anomalies; Coronary artery; Data; Development; Developmental Process; EDN1 gene; Endothelin-1; Endothelium; Environment; Enzymes; Extracellular Matrix; Fetal Development; First Generation College Students; Funding; Future; Goals; Heart; In Vitro; Injections; Knowledge; Lead; Legal patent; Life; Liquid substance; Location; MMP2 gene; MMP9 gene; Metalloproteases; Minor; Molecular; Monitor; Muscle; Muscle Proteins; NOS3 gene; Nitric Oxide Synthase; Pattern; Penetration; Peptide Signal Sequences; Phosphorylation; Physicians; Process; Production; Proliferating; Research; Research Personnel; Research Proposals; Role; Scientist; Signal Induction; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Students; Sudden Death; Techniques; Time; Tissues; Universities; Variant; Vertebrates; Visualization; Work; base; cell behavior; cell motility; coronary vasculature; experimental study; inhibitor; migration; prevent; protein expression; recruit; response; shear stress; spatiotemporal; transcription factor; undergraduate research; undergraduate student

Project Summary The broad objective of this proposal is to understand why the developing coronary arteries are consistently formed at specific locations. During development of the coronary arteries, numerous immature coronary strands connect to the aorta and then remodel to form the two mature arteries observed in the adult heart. During this remodeling process, one of the biggest changes applied to the newly connected coronary vasculature is the sudden onset of fluid shear stress, yet the regulatory role of fluid shear stress on the development of the coronary vasculature is completely unknown. This proposal specifically addresses how the onset of blood flow, which causes shear stress, induces signaling pathways that would promote smooth muscle migration, proliferation, and maturation. Aim 1 examines the environment surrounding the aorta during the early stages of smooth muscle recruitment to the coronaries. Low levels of shear stress are predicted to promote expression of ET1 and thus lead to matrix metalloprotease (MMP) activity, yielding an environment that promotes smooth muscle migration. This aim will evaluate ET1, MMP2, and MMP9 expression as well as smooth muscle proliferation in the developing coronary arteries of the chick embryo. In ovo injection of MMP2 and 9 inhibitors will confirm the necessity of these MMPs for smooth muscle migration. Aim 2 examines the effects of increased levels of shear stress on cell signaling, particularly the transcription factor KLF2 and the nitric oxide synthase eNOS. High levels of shear stress are predicted to induce KLF2 and lead to eNOS phosphorylation, which promote smooth muscle maturation. Asymmetrical shear stress would thus selectively mature smooth muscle, explaining the consistent patterning of the coronary arteries. In ovo inhibition of eNOS activity will confirm whether eNOS activity is required for smooth muscle maturation and coronary artery stability. Most of the proposed techniques are easily accessible to undergraduate research students, and the preliminary data were generated by undergraduate students, supporting the feasibility of this project at the University of Portland. Altogether, this proposal sits at the intersection among physical forces, molecular responses, and cellular behavior, with the end objective of understanding the formation of the coronary arteries. By better understanding the normal development of these arteries, the proposed research will support future work to examine how these developmental processes go awry, leading to life-threatening congenital coronary artery anomalies.

项目摘要 该提议的广泛目标是了解为什么发展的冠状动脉一直是一贯的 在特定位置形成。在冠状动脉发展过程中，许多未成熟的冠状动脉 链连接到主动脉，然后重塑以形成在成年心脏中观察到的两个成熟动脉。 在此重塑过程中，最大的变化之一适用于新连接的冠状动脉 脉管系统是液体剪切应力的突然发作，但是流体剪切应力在 冠状动脉脉管系统的发展是完全未知的。该建议专门解决了如何 血流的发作，导致剪切应力，引起信号传导途径，以促进平滑肌 迁移，增殖和成熟。 AIM 1检查了主动脉周围的环境 平滑肌招募冠状动脉的早期阶段。预计低水平的剪切应力将 促进ET1的表达，从而导致基质金属蛋白酶（MMP）活性，产生环境 这促进了平滑的肌肉迁移。此目的将评估ET1，MMP2和MMP9表达以及 雏鸡胚胎发育中的冠状动脉中的平滑肌增殖。在OVO注入MMP2中 和9个抑制剂将确认这些MMP的平滑肌迁移的必要性。 AIM 2检查 剪切应力水平增加对细胞信号的影响，特别是转录因子KLF2和 一氧化氮合酶eNOS。预计高水平的剪切应力会诱导KLF2并导致eNOS 磷酸化，促进平滑肌成熟。因此，不对称剪切应力将选择性地选择性 成熟的平滑肌，解释了冠状动脉的一致图案。在OVO抑制eNOS中 活动将确认是否需要平滑肌肉成熟和冠状动脉活动 稳定。大多数建议的技术很容易被本科研究专业的学生访问， 初步数据是由本科生生成的，支持该项目的可行性 波特兰大学。总共，该提议位于物理力之间的交集，分子 反应和细胞行为，最终目标是理解冠状动脉的形成 动脉。通过更好地了解这些动脉的正常发展，拟议的研究将支持 未来的工作，以检查这些发展过程如何出现问题，从而导致先天性威胁生命 冠状动脉异常。