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: 10630420
• 负责人: Yibing Qyang
• 金额: $ 6.38万
• 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/10630420
• 关键词: Address; Age; Autologous; Behavior; Bioreactors; Caliber; Caring; Cell Line; Cell physiology; Cells; Data; Development; Disease; Due Process; Endothelial Cells; Endothelium; Genes; Goals; Human; Immunocompromised Host; Implant; Modeling; Operative Surgical Procedures; Patients; Polyglycolic Acid; Procedures; Radioisotopes; Rattus; Research; Research Personnel; SLC5A5 gene; Smooth Muscle Myocytes; Technology Assessment; Thrombosis; Time; Training; Validation; Vascular Graft; Vascular Smooth Muscle; Visualization; Work; X-Ray Computed Tomography; base; endothelial stem cell; health assessment; implantation; in vivo; induced pluripotent stem cell; injury and repair; innovative technologies; parent grant; prevent; repaired; shear stress; single photon emission computed tomography; solute; stem cell derived tissues; stem cells; technology development; time use; tissue stem cells; transcription activator-like effector nucleases; uptake; vascular injury; vascular tissue engineering

Project Summary Tissue engineered vascular grafts (TEVGs) produced through seeding human induced pluripotent stem cell (hiPSC)-derived vascular smooth muscle cells (VSMCs) onto polyglycolic acid (PGA) meshes have been developed to repair patients’ vascular injuries. These hiPSC- TEVGs can be made further applicable through decellularization, allowing them to be stored for long periods and used off the shelf. For large diameter vessels (>6mm inner diameter), these decellularized TEVGs can be implanted directly, with host cells sufficiently able to recellularized the graft over time. However, small diameter TEVGs (2-4mm) are prone to thrombosis if implanted directly, requiring a layer of endothelial cells (ECs) to be coated in the lumen before these procedures. Significant research has been done to create hiPSC-ECs sufficient for endothelialization of small diameter TEVGs, as many patients have autologous ECs unable to be used for this process due to age or disease. For this reason, it is vital for the development of this technology for researchers to know how these coated hiPSC-ECs behave in vivo non- invasively. In this supplemental proposal, this need will be addressed by producing a stable hiPSC line expressing human sodium iodide symporter (hNIS), a solute carrier that confers the ability to uptake radioisotopes into the cell. These hNIS-hiPSCs will be used to derive ECs that can be tracked in vivo non-invasively using SPECT/CT imaging, providing key data to the behavior and function of these cells after implantation. This will be accomplished through the use of transcription activator-like effector nuclease (TALEN) gene editing to insert a constitutively active hNIS cassette into the AAVS1 “safe harbor” locus in stable hiPSCs. hiPSC- TEVGs will then be decellularized and coated with hNIS-ECs derived from the hNIS-hiPSCs, and these grafts will be trained for implantation by undergoing shear stress in a flow bioreactor, to enhance hNIS-EC maturity. These matured hNIS-EC TEVGs will then be used as a aortic interposition grafts in an immunocompromised rat model, which allow for the validation of the functionality of these hNIS-ECs to allow in vivo visualization over time using SPECT/CT imaging. This innovative technology will allow for non-invasive assessment of the health and extent of implanted hiPSC-EC coating in decellularized grafts, providing vital information into the ability of the endothelium to prevent thrombosis in patients who require vascular injury repair.

项目概要 通过播种人体诱导产生的组织工程血管移植物（TEVG） 将多能干细胞 (hiPSC) 来源的血管平滑肌细胞 (VSMC) 移植到聚乙醇酸上 酸性（PGA）网已被开发用于修复患者的血管损伤。 TEVG 可以通过去细胞化进一步应用，从而可以将它们储存起来以供使用。 对于大型容器（内径>6mm），这些是长期使用的。 脱细胞TEVG可以直接植入，宿主细胞足以再细胞化 然而，随着时间的推移，小直径 TEVG（2-4 毫米）很容易形成血栓。 直接植入，需要在管腔内涂覆一层内皮细胞​​（EC） 这些程序已经进行了大量的研究来创建足以用于的 hiPSC-EC。 小直径 TEVG 的内皮化，因为许多患者具有自体 EC，无法 由于年龄或疾病而用于此过程因此，它对于发育至关重要。 这项技术让研究人员能够了解这些包被的 hiPSC-EC 在体内的非 在这个补充提案中，这一需求将通过生产稳定的解决方案来解决。 hiPSC 系表达人碘化钠同向转运体 (hNIS)，这是一种溶质载体，可赋予 这些 hNIS-hiPSC 将用于衍生 EC 可以使用 SPECT/CT 成像进行体内非侵入性追踪，为研究人员提供关键数据 这些细胞在植入后的行为和功能将通过 使用转录激活因子样效应核酸酶 (TALEN) 基因编辑来插入 将组成型活性 hNIS 盒插入稳定 hiPSC 中的 AAVS1“安全港”位点。 然后 TEVG 将被脱细胞并涂有源自 hNIS-hiPSC 的 hNIS-EC， 这些移植物将通过在流动生物反应器中承受剪切应力来进行植入训练， 以提高 hNIS-EC 的成熟度。这些成熟的 hNIS-EC TEVG 将被用作主动脉。 在免疫功能低下的大鼠模型中进行插入移植物，这可以验证 这些 hNIS-EC 的功能允许使用 SPECT/CT 进行体内可视化 这项创新技术将允许对健康和健康进行非侵入性评估。 脱细胞移植物中植入 hiPSC-EC 涂层的程度，为移植物提供重要信息 内皮细胞预防需要血管损伤修复的患者血栓形成的能力。