CAREER: Vascular Stent Geometry Dictates Endothelial Cell Wound Healing and Phenotype

心脏病，包括冠状动脉疾病，是美国和世界范围内的主要死亡原因。 冠状动脉疾病是由动脉粥样硬化阻塞心脏的一条或多条血管引起的，动脉粥样硬化是斑块的积聚，可导致心脏病发作。 支架通常用于治疗冠状动脉疾病患者以防止血管闭合。 然而，支架也带来临床并发症，通常与引入支架后在血管中产生的伤口的延迟愈合有关。 内皮细胞是血管内部的细胞，在将支架引入血管后的伤口愈合中起关键作用。该教师早期职业发展计划（CAREER）项目的目标是研究单个支架支柱周围的血流环境如何影响内皮细胞，从而促进或阻止伤口愈合。 具体来说，该项目将研究如何使用分子生物学和工程学相结合的方法来研究内皮细胞基因的表达受到支架支柱附近血流的影响。 这一知识的进步将支持开发新的治疗方法，以加速引入支架后的伤口愈合，并减少与支架相关的并发症。 教育和推广活动将与实验室研究相结合。 该奖项还将为从小学到本科和研究生的不同教育水平的学生提供科学、技术、工程和数学（STEM）的机会。总体研究目标是确定支架周围流体流动环境中内皮细胞伤口愈合的遗传基础，这是为支架术后患者开发靶向治疗的关键一步。 该奖项研究的假设是支架支柱周围的流体流动环境阻碍了由内皮细胞指导的正常伤口愈合过程。 这将通过三个研究目标得到支持。 在目标1中，该项目将在模拟支架的体外环境中识别出阻止或促进伤口愈合的基因。 在目标2中，该项目将开发一种独特的高通量流体流动系统，该系统将能够同时测试多个基因和流动条件，快速推进结果的收集。 目标3将通过使用CRISPR/Cas9介导的敲除/敲入直接修饰内皮细胞中的基因表达以拯救定向细胞迁移并促进伤口愈合来直接测试参与内皮细胞定向伤口愈合的基因。 该项目还包括若干教育目标。 大学生将通过课程接触到工程和医学的融合，课外实验室模块将有助于提高对跨学科STEM职业的认识。 该奖项还将扩大现有的STEM试点项目，面向在家上学的小学生，这是一个经常被忽视但在美国不断增长的学生群体，为了开发一个互补的科学课程，目标是达到其他在家上学的社区在国家和全国范围内。这个奖项反映了NSF的法定使命，并已被认为是值得通过评估使用基金会的智力价值和更广泛的影响支持审查标准。

Heart disease, including coronary artery disease, is the leading cause of death in both the USA and worldwide. Coronary artery disease is caused by blockage of one or more blood vessels of the heart by atherosclerosis, the build-up of plaques, which can lead to a heart attack. Stents are commonly used to treat coronary artery disease patients to prevent closure of the blood vessel. However, stents also carry clinical complications, often related to delayed healing of the wound that is created in the blood vessel after the stent is introduced. Endothelial cells, the cells lining the inside of blood vessels, play a key role in wound healing after a stent is introduced into the blood vessel. The goal of this Faculty Early Career Development Program (CAREER) project is to investigate how the blood flow environment around the individual stent struts affects endothelial cells so as to promote or prevent wound healing. Specifically, the project will study how expression of endothelial cell genes is affected by blood flow adjacent to the stent struts using a combined molecular biology and engineering approach. This advancement in knowledge will support the development of new treatment approaches to accelerate wound healing after introduction of the stent and decrease the complications associated with stents. Education and outreach activities will be integrated with the laboratory research. This award will also foster Science, Technology, Engineering and Math (STEM) opportunities at different educational levels, ranging from elementary school to undergraduate and graduate students.The overall research objective is to identify the genetic basis of endothelial cell wound healing in the fluid flow-environment surrounding a stent, a crucial step toward developing targeted therapies for patients after stenting. The hypothesis being investigated through this award is that the fluid flow environment around the stent strut impedes the normal wound healing process that is directed by endothelial cells. This will be supported through three research aims. In aim 1, the project will identify genes that prevent or promote wound healing in an in vitro environment that simulates stents. In aim 2, the project will develop a unique, high-throughput fluid flow system that will enable testing of multiple genes and flow conditions simultaneously, rapidly advancing the collection of results. Aim 3 will directly test the genes involved in endothelial cell-directed wound healing by using CRISPR/Cas9 mediated knock-out/knock-in to directly modify gene expression in endothelial cells to rescue directed cell migration and promote wound healing. The project also includes several educational objectives. University students will be exposed to the integration of engineering and medicine through courses, and extracurricular laboratory modules will contribute to increased awareness of interdisciplinary STEM careers. The award will also expand an existing pilot program in STEM for homeschooled elementary students, an often overlooked but rising student population in the US, in order to develop a complementary science curriculum with the goal to reach other homeschooling communities in the state and nationwide.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.