Readily Available Stem Cell-Based Vascular Grafts for Emergent Surgical Care

用于紧急手术护理的现成干细胞血管移植物

• 批准号: 10414459
• 负责人: Yibing Qyang
• 金额: $ 6.19万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10414459
• 关键词: BCL2 gene; Caliber; Caring; Cell Line; Cell physiology; Cells; Data; Development; Disease; Doxycycline; Elderly; Endothelial Cells; Endothelium; Ensure; Event; Genes; Goals; Human; Immunocompromised Host; Implant; Lead; Liquid substance; Modeling; Operative Surgical Procedures; Patients; Polyglycolic Acid; Population; Problem Solving; Rattus; Research Personnel; Risk; Safety; Smooth Muscle Myocytes; Source; System; Thrombosis; Thymidine Kinase; Time; Tissues; Training; Traumatic injury; Vascular Graft; Vascular Smooth Muscle; Work; acute care; base; clinical application; efficacy testing; implantation; induced pluripotent stem cell; inducible gene expression; injury and repair; parent grant; patient population; repaired; scaffold; shear stress; stem cells; thrombotic; transcription activator-like effector nucleases; vascular injury; vascular tissue engineering

Project Summary Tissue engineered vascular grafts (TEVGs) for traumatic vascular injury repair in small diameter (2- 4mm) vessels can be made from human induced pluripotent stem cell derived vascular smooth muscle cells (hiPSC-VSMC) seeded onto a polyglycolic acid (PGA) scaffold. Furthermore, these TEVGs can be subsequently decellularized and stored long term for use in acute care for traumatic injury. For larger diameter vessel repair, it is adequate to implant an acellular scaffold and allow host cells to migrate from anastomotic edges to line the implanted vessel lumen. However, this model is not effective for smaller diameter vessels that are more prone to thrombosis. Researchers have attempted to solve this problem by coating the lumen of implanted vessels with endothelial cells (ECs) prior to implantation. However, data from implanted grafts suggests there is a short turnover time for implanted EC populations within these grafts. In the case of elderly or diseased patients, they may not retain quality EC function. Therefore, this patient population may display poor integration of host cells into the implanted tissue, which may lead to an increased risk of thrombotic and stenotic events. The purpose of this project is to generate a stable hiPSC line with controllable expression of the pro-survival factor Bcl-2 to generate hiPSC derived endothelial cells (hiPSC-ECs) as a readily available cell source to line the lumen of decellularized TEVGs prior to implantation. A stable hiPSC line with doxycycline inducible expression of the pro-survival factor Bcl-2 will be generated via transcription activator-like effector nucleases (TALEN) gene editing. Additionally, a robust “safety switch” system using ectopically expressed thymidine kinase (TK) will be employed to ensure cells may be removed post implantation should unwanted effects occur. Decellularized TEVGs will have their lumen coated with hiPSC-ECs made from this gene edited Bcl-2+TK+ cell line. These endothelialized TEVGs will undergo fluid shear stress training to enhance hiPSC-EC function prior to implantation into an immunocompromised rat model as an aortic interposition graft to test the efficacy of this doxycycline inducible Bcl-2 system. This platform will allow for a readily available cell line to be used to produce a long lasting hiPSC-EC lumen that can maintain patency of the graft as it fully integrates with patients who may have subpar EC function.

项目概要 用于修复小直径创伤性血管损伤的组织工程血管移植物（TEVG）（2- 4mm）血管可以由人类诱导多能干细胞衍生的血管平滑制成 肌肉细胞（hiPSC-VSMC）接种到聚乙醇酸（PGA）支架上此外，这些。 TEVG 随后可以脱细胞并长期保存，用于创伤性急性护理 对于较大直径的血管修复，植入无细胞支架并允许宿主就足够了。 细胞从吻合边缘迁移到植入的血管腔内。 研究人员发现，对于更容易形成血栓的较小直径血管无效。 试图通过用内皮细胞涂覆植入血管的管腔来解决这个问题 (EC) 植入前然而，植入移植物的数据表明存在短暂的周转。 对于老年或患病患者，在这些移植物中植入 EC 群体的时间。 他们可能无法保留高质量的 EC 功能，因此，该患者群体可能表现不佳。 宿主细胞整合到植入的组织中，这可能导致血栓形成的风险增加 该项目的目的是生成具有可控性的稳定 hiPSC 系。 表达促生存因子 Bcl-2 以产生 hiPSC 衍生内皮细胞 (hiPSC-EC) 作为一种现成的细胞来源，可在植入前排列脱细胞 TEVG 的管腔。 具有多西环素诱导表达促生存因子 Bcl-2 的稳定 hiPSC 系将是 通过类转录激活因子效应核酸酶 (TALEN) 基因编辑生成。 使用异位表达的胸苷激酶（TK）的强大“安全开关”系统将用于 确保在发生不良影响时可以在植入后去除细胞。 TEVG 的管腔将涂有由该基因编辑的 Bcl-2+TK+ 细胞系制成的 hiPSC-EC。 这些内皮化 TEVG 将接受流体剪切应力训练，以增强 hiPSC-EC 功能 在作为主动脉介入移植物植入免疫功能低下的大鼠模型之前，以测试 该多西环素诱导 Bcl-2 系统的功效该平台将允许容易获得的细胞。 用于产生持久的 hiPSC-EC 管腔的生产线，可以保持移植物的通畅，因为它 与 EC 功能可能低于标准的患者完全融合。