Endothelial Function in Response to Topographical Micropatterning

内皮功能对地形微图案的反应

• 批准号: 10535346
• 负责人: Meghan Elizabeth Fallon
• 金额: $ 4.68万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-22 至 2025-08-21
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10535346
• 关键词: Actins; Adult; Affect; Anastomosis - action; Anti-Inflammatory Agents; Attenuated; Autologous Transplantation; Biocompatible Materials; Biological Response Modifiers; Blood; Bypass; Caliber; Cardiovascular Diseases; Cause of Death; Cell Adhesion Molecules; Cell Shape; Cell physiology; Cells; Cellular Morphology; Chronic; Clinical; Complex; Cues; Data; Development; Down-Regulation; Endothelial Cells; Endothelium; Environment; Equilibrium; Exposure to; Failure; Functional disorder; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Goals; Growth; Hemostatic Agents; Hemostatic function; In Vitro; Individual; Inflammatory; Intervention; Liquid substance; Mechanical Stress; Mechanoreceptors; Mediating; Mediator of activation protein; Microtubule Polymerization; Microtubules; Modification; Morphology; Pathway interactions; Patient-Focused Outcomes; Patients; Pattern; Phenotype; Procedures; Proteins; Regulation; Resistance; Role; Side; Signal Pathway; Surface; TNF gene; Techniques; Technology; Thrombosis; Thrombus; Tissue-Specific Gene Expression; Tissues; Transducers; Up-Regulation; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Cell; Vascular Graft; Western Blotting; Work; biomaterial compatibility; cell growth; cytokine; design; graft function; hemodynamics; high risk; immunogenic; immunogenicity; improved; inflammatory milieu; monolayer; mortality; novel; protein activation; protein expression; response; shear stress; tool; transcription factor; transcriptome sequencing

Project Summary Cardiovascular disease affects nearly half of all U.S. adults and is the leading cause of death worldwide. Advanced cases are often treated through vascular grafting to bypass occluded vessels. Synthetic vasculargraft materials suffer from patency complications due to thrombosis and neointimal growth impeding the materials’ long-term function for small-diameter applications. Thus, there is a critical unmet need for improved biocompatible small-diameter vascular grafts in order to support long-term patient outcomes and reduce re- intervention procedures. The in vitro establishment of an endothelial layer on synthetic biomaterials has been suggested to be a solution due to the endothelial cells’ (ECs) homeostatic capabilities to prev ent thrombus formation and limit immunogenicity. Therefore, vascular graft material surfaces which support EC growth and function are a significant clinical need. ECs actively respond to both material surface cues and local hemodynamic fluid shear stress (FSS) to balance hemostasis, immuno-protection, and thrombo-resistance via a variety of mechano-receptors and -transducers. While unidirectional FSS induces morphological whole cell elongation and alignment as well as alignment of cytoskeletal components, oscillatory FSS (located at end-to- side anastomoses of vascular grafts) induces a cobblestoneEC morphology with randomcytoskeletal alignment. Krüppel-like factor 2 (KLF2) and Yes-associated protein (YAP) are transcription factors that are highly sensitive to cell shape and mechanical stresses. KLF2 and YAP have been shown to regulate EC function and phenotype. KLF2 is upregulated under unidirectional FSS, resulting in an anti-inflammatory phenotype. YAP is hyper- activated under oscillatory FSS and induces an immuno-prone phenotype, while remaining inactivated under unidirectional FSS. However, in the absence of FSS, the role of transcription factor regulation in EC morphology and cytoskeletal alignment driven immuno-protection is unknown. EC morphology and cytoskeletal alignment can be controlled using techniques of topographical micropatterning, independent of hemodynamic flow. We and others have shown that topographical micropatterning drivesan anti-inflammatory EC phenotypein static culture, making it a promising tool for synthetic graft surfaces. Studies of micropatterned ECs have shown additive benefits of patterning to unidirectional FSS and the ability to maintain cellular elongation under orthogonal unidirectional flow. This suggests that ECs on micropatterned surfaces may be more resistant to the immunogenic effects of oscillatory flow; yet this has never been directly studied. The proposed work aims to (1) elucidate the mechanism by which transcription factors regulate endothelial morphology driven functions independent of hemodynamic effects and (2) determine the effect of oscillatory flow on micropatterned EC transcription factor regulation and immunogenicity. Determining how ECs respond to topographical micropatterning is critical in designing improved synthetic vascular grafts for small-diameter applications.

项目摘要 心血管疾病几乎影响了所有美国成年人的一半，并且是全球死亡的主要原因。 经常通过血管嫁接以绕过封闭的血管来治疗晚期病例。合成血管移植 由于血栓形成和新内膜生长引起的材料并发症遭受了通知并发症的影响，从而阻碍了材料 小直径应用的长期功能。那是一个至关重要的改进需求 生物相容性的小直径血管移植物，以支持长期的患者预后 干预程序。体外建立在合成生物材料上的内皮层一直是 由于内皮细胞（ECS）的稳态功能，建议是一种解决方案 形成和限制免疫原性。因此，支持EC生长和 功能是巨大的临床需求。 EC对材料表面提示和局部的反应 血液动力学剪切应力（FSS）以平衡止血，免疫保护和抗血栓抗性 多种机械振动器和 - 转移器。而单向FSS诱导形态学全细胞 伸长和比对以及细胞骨架成分的振荡FSS（位于端到 - 血管移植物的侧吻合）诱导具有随机骨骨骼比对的鹅卵石形态。 Krüppel样因子2（KLF2）和与YES相关蛋白（YAP）是高度敏感的转录因子 到细胞形状和机械应力。 KLF2和YAP已显示用于调节EC功能和表型。 KLF2在单向FSS下进行更新，从而导致抗炎表型。 yap是超级 在振荡的FSS下激活并诱导免疫易发的表型，同时仍在灭活下 单向FSS。但是，在没有FSS的情况下，转录因子调节在EC形态中的作用 细胞骨架比对驱动免疫保护尚不清楚。 EC的形态和细胞骨架比对 可以使用与血流动力学无关的地形微观图案的技术来控制。我们和 其他人则表明，地形微图案驱动器抗炎EC型表型静态培养， 使其成为合成移植表面的有前途的工具。对微图案EC的研究添加了 对单向FSS的图案效果以及在正交下保持细胞伸长的能力 单向流。这表明微图表上的EC可能对 振荡流的免疫原性；然而，这从来没有直接研究。拟议的工作旨在（1） 阐明转录因子调节内皮形态驱动功能的机制 独立于血液动力学作用，（2）确定振荡流对微图案EC的影响 转录因子调节和免疫原性。确定EC对地形的反应 微图对于设计改进的小直径应用的合成血管移植物至关重要。