Transcriptomics of adherent endothelial cells for improved endothelialization of small-diameter vascular grafts

项目摘要 这项研究的长期目标是在血液接触表面建立自体内皮细胞 可植入的心血管设备。这对于提供血液相容性和最大限度地减少 降低血栓形成、栓塞等不良后果的风险，以提高患者的安全性。 这一目标直接支持了NHLBI促进心脏病治疗和 增进所有人的健康，使他们能够活得更长、更有成效。许多心血管疾病 疾病是用植入式设备治疗的。在拯救生命的同时，这些设备也存在固有的风险 需要进行抗凝治疗的血栓形成和血栓形成。 拟议的研究解决了在小直径内建立内皮细胞的迫切临床需求。 血管移植。需要接受搭桥手术的冠心病患者目前正在接受自体心脏移植 血管收获，因为合成移植物在小直径应用中表现出不佳的通畅性。自体 血管收获与额外的成本和显著的供体部位发病率有关。此外，一个大型和 越来越多的患者由于包括先前存在的血管在内的原因而没有合适的自体血管 疾病、脉络剥离和以前的收成。小直径自体内皮细胞的建立 要达到可接受的通畅率，而不需要自体血管收获，搭桥是必要的。 这项拟议的研究的目标是通过以下方式改善内皮细胞在血管移植生物材料上的保留率 为促进细胞黏附的分子调控策略寻找新的分子信号靶点 力量。这是由我们的假设推动的，即分子信号通路的不同调节涉及 细胞黏附是内皮细胞在植入后抵抗脱离的原因。 具体目标1将确定负责允许内皮细胞亚群的分子信号通路 细胞暴露在生理剪应力下时保持粘连。RNA测序将用于比较 所有细胞和贴壁细胞的转录组。转录组数据将用于鉴定分子 在贴壁细胞亚群中高度差异调节的信号。 特殊目标2将开发和测试改善血管移植物上内皮细胞留存的策略 生物材料。将开发一种分子调控策略，通过上调OR来增强细胞黏附 适当下调关键信号分子。细胞保留率将在以下条件下进行比较 生理剪应力。 特殊目标3将评估种植内皮细胞的小直径血管移植物的生物学性能 使用临床前模型的细胞。最有希望的细胞黏附增强策略将用于 将内皮化移植物植入猪的颈动脉循环并进行临床分析 相关结果包括细胞滞留、通畅、血栓形成、新生内膜增生和炎症。

Project Summary The long-term objective of this study is to establish an autologous endothelium on the blood-contacting surface of implantable cardiovascular devices. This is critically important for providing hemocompatibility and minimizing the risk of thrombosis, embolism, and other adverse outcomes in order to improve patient safety. This objective directly supports the mission of the NHLBI to promote the treatment of heart diseases and enhance the health of all individuals so that they can live longer and more productive lives. Many cardiovascular diseases are treated with implantable devices. While lifesaving, these devices also carry an inherent risk of thrombosis and embolism that requires administration of anticoagulation therapy. The proposed study addresses the dire clinical need for establishing an endothelium within small-diameter vascular grafts. Patients with coronary heart disease who require bypass surgery currently undergo autologous vessel harvest because synthetic grafts demonstrate poor patency in small-diameter applications. Autologous vessel harvest is associated with additional cost and significant donor site morbidity. In addition, a large and growing number of patients do not possess suitable autologous vessel for reasons including preexisting vascular disease, vein stripping, and previous harvest. Establishment of an autologous endothelium on small-diameter bypass grafts is necessary for achieving acceptable patency rates without autologous vessel harvest. The goal of the proposed research is to improve endothelial cell retention on vascular graft biomaterials by identifying novel molecular signaling targets for molecular modulation strategies to promote cellular adhesion strength. This is driven by our hypothesis that differential regulation of molecular signaling pathways involved in cellular adhesion is responsible for a subpopulation of endothelial cells resisting detachment upon implantation. Specific Aim 1 will identify molecular signaling pathways responsible for allowing a subpopulation of endothelial cells to remain adherent upon exposure to physiological shear stress. RNA-sequencing will be used to compare the transcriptome of all cells and adherent cells. The transcriptomics data will be used to identify molecular signals that are highly differentially regulated in adherent cell subpopulations. Specific Aim 2 will develop and test strategies to improve endothelial cell retention on vascular graft biomaterials. A molecular modulation strategy will be developed to enhance cell adhesion by upregulating or downregulating critical signaling molecules as appropriate. Cell retention will be compared under conditions of physiological shear stress. Specific Aim 3 will assess the biological performance of small-diameter vascular grafts seeded with endothelial cells using a preclinical model. The most promising cell adhesion enhancement strategy will be used to endothelialize grafts, which will be implanted into the carotid circulation of swine and analyzed for clinically relevant outcomes including cell retention, patency, thrombosis, neointimal hyperplasia, and inflammation.