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

贴壁内皮细胞的转录组学用于改善小直径血管移植物的内皮化

• 批准号: 10365253
• 负责人: Brandon J Tefft
• 金额: $ 39.15万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10365253
• 关键词: Address; Animal Model; Anticoagulation; Area; Autologous; Biocompatible Materials; Biological; Blood; Blood Circulation; Blood flow; Bypass; CRISPR/Cas technology; Caliber; Cardiovascular Diseases; Cardiovascular system; Cell Adhesion; Cell physiology; Cells; Clinical; Coronary; Coronary Artery Bypass; Coronary heart disease; Data; Development; Devices; Embolism; Endothelial Cells; Endothelium; Engineering; Environment; Exposure to; Family suidae; Gene Activation; Gene Expression Profile; Generations; Genes; Goals; Harvest; Health; Heart Diseases; Heart Valves; Hyperplasia; Implant; Individual; Inflammation; Inflammatory Response; Liquid substance; Maintenance; Mediating; Medicine; Mission; Molecular; Molecular Target; Morbidity - disease rate; National Heart, Lung, and Blood Institute; Operative Surgical Procedures; Outcome; Patients; Performance; Phenotype; Physiological; Population; Positioning Attribute; Pre-Clinical Model; Regenerative Medicine; Regenerative engineering; Regulation; Research; Risk; Safety; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Small Interfering RNA; Stents; Surface; TNF gene; Testing; Thrombosis; Tissue Engineering; Vascular Diseases; Vascular Graft; Veins; adverse outcome; base; biomaterial compatibility; carcinoembryonic antigen-related cell adhesion molecules; cell behavior; clinically relevant; cost; design; hemocompatibility; hemodynamics; implantable device; implantation; improved; insight; knock-down; nanoscale; novel; overexpression; patient safety; rational design; risk minimization; shear stress; transcriptome; transcriptome sequencing; transcriptomics

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.

项目概要 本研究的长期目标是在血液接触表面建立自体内皮细胞 植入式心血管装置。这对于提供血液相容性并最大限度地减少 血栓形成、栓塞和其他不良后果的风险，以提高患者安全。 这一目标直接支持 NHLBI 促进心脏病和心脏病治疗的使命 增强所有人的健康，使他们能够活得更长久、更富有成效。许多心血管 疾病通过植入设备进行治疗。在挽救生命的同时，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栓塞。 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 血管移植物。需要搭桥手术的冠心病患者目前正在接受自体移植手术 血管收获，因为合成移植物在小直径应用中表现出通畅性差。自体 血管采集与额外成本和显着的供体部位发病率相关。此外，还有一个大而 由于已有血管等原因，越来越多的患者没有合适的自体血管 疾病、静脉剥离和之前的收获。小直径自体内皮细胞的建立 旁路移植对于在不采集自体血管的情况下获得可接受的通畅率是必要的。 拟议研究的目标是通过以下方式改善血管移植生物材料上的内皮细胞保留： 确定分子调节策略的新分子信号传导靶标，以促进细胞粘附 力量。这是由我们的假设驱动的，即分子信号通路的差异调节涉及 细胞粘附导致内皮细胞亚群在植入时抵抗脱离。 具体目标 1 将确定负责允许内皮细胞亚群形成的分子信号传导途径 细胞在暴露于生理剪切应力时保持贴壁。 RNA测序将用于比较 所有细胞和贴壁细胞的转录组。转录组学数据将用于鉴定分子 在贴壁细胞亚群中受到高度差异调节的信号。 具体目标 2 将开发和测试改善血管移植物上内皮细胞保留的策略 生物材料。将开发一种分子调节策略，通过上调或 适当下调关键信号分子。将在以下条件下比较细胞保留 生理剪切应力。 具体目标 3 将评估接种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。最有前途的细胞粘附增强策略将用于 内皮化移植物，将其植入猪的颈动脉循环中并进行临床分析 相关结果包括细胞滞留、通畅、血栓形成、新内膜增生和炎症。