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: 10189694
• 负责人: Yibing Qyang
• 金额: $ 66.43万
• 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/10189694
• 关键词: Abdomen; Acute; Address; Allogenic; Anastomosis - action; Animal Model; Animals; Arteries; Autologous; Autologous Transplantation; Biomechanics; Bioreactors; Blast Injuries; Blood; Blood Vessels; Blood coagulation; Blunt Trauma; Bypass; Caliber; Cardiovascular Physiology; Caring; Cells; Clinical; Clothing; Coagulation Process; Collagen; Contusions; Cryopreservation; Deposition; Derivation procedure; Diabetes Mellitus; Disease; Ectopic Expression; Elastin; Elderly; Emergency Situation; Emergency treatment; Endothelial Cells; Endothelium; Engineering; Ethical Issues; Extracellular Matrix; Family suidae; Fracture; Functional disorder; Future; Generations; Growth; HLA Antigens; Hemodialysis; Histologic; Human; Immune response; Implant; In Vitro; Injury; Intervention; Lead; Letters; Life; Ligation; Limb structure; Lower Extremity; Mechanics; Modeling; Monitor; Morbidity - disease rate; Nature; Nude Rats; Operative Surgical Procedures; Oxygen; Patient Care; Patients; Performance; Periodicity; Peripheral; Phase II Clinical Trials; Polyglycolic Acid; Pre-Clinical Model; Production; Property; Proteins; Rattus; Reproducibility; Residual state; Saphenous Vein; Smooth Muscle Myocytes; Somatic Cell; Source; Stem Cell Factor; Stretching; Supplementation; Surface; Tensile Strength; Tibial Arteries; Time; Traffic accidents; Transplantation; Trauma; Treatment Efficacy; Variant; Vascular Endothelial Cell; Vascular Graft; Vascular Smooth Muscle; Vascular remodeling; Wait Time; advanced disease; base; brachial artery; cell type; endothelial stem cell; experience; immunogenic; immunosuppressed; implantation; in vivo; in vivo Model; induced pluripotent stem cell; infection risk; injured; injury and repair; limb loss; mechanical properties; morphometry; mortality; patient population; porcine model; pre-clinical; preclinical efficacy; prevent; scaffold; scale up; severe injury; stem cell derived tissues; stem cells; thrombogenesis; tissue stem cells; urgent care; vascular injury; vascular tissue engineering; virtual

Patients who have undergone vascular trauma resulting from penetrating (e.g. blast injury) or blunt (e.g. fracture and contusion from a traffic accident) trauma often experience severe damage to their small-diameter peripheral arteries (2-4mm, e.g. tibial or brachial arteries) and require surgical intervention to bypass or replace the injured vessel segments. As the accessibility of saphenous veins may be low in patients of vascular trauma due to severe injuries or loss of more than one lower extremity as a result of blast injury or severe traffic accident, and the application of synthetic vascular grafts can be hindered by the potential risk of infection due to the contagious nature of these types of injuries, an alternative source of vascular graft is in dire need. Acellular tissue engineered vascular grafts (TEVGs) therefore may offer a readily available treatment for emergency care for patients of vascular trauma. Although TEVGs of significant mechanical strength can be developed from culturing primary human vascular smooth muscle cells (VSMCs) followed by decellularization, this platform has several obstacles which prevent it from ultimately serving victims of vascular trauma. The finite expandability, limited accessibility, and donor-to-donor functional variations among primary VSMCs may hinder the efficiency of TEVG production. Further, as small-diameter TEVGs require autologous endothelial cell (EC) coating prior to implantation to avoid blood clothing, potential dysfunction of patient ECs due to either advanced age or disease, as well as the significant time required to derive and expand the patient's autologous ECs, could prevent applicability in treating acute vascular injury. Human induced pluripotent stem cells (hiPSCs), self-renewable cells derived from somatic cells by the ectopic expression of stem cell factors, provide an excellent alternative to address the complications of primary cell based TEVGs. As hiPSCs can be differentiated into virtually any somatic cell type, including VSMCs and ECs (hiPSC-VSMCs and hiPSC-ECs), these cells provide an unlimited cell source to obtain vascular cells to construct TEVGs of comparable quality (hiPSC-TEVGs). Further, by engineering the expression of human leukocyte antigen (HLA) proteins, non- or low-immunogenic universal hiPSCs could be established, making the hiPSC-TEVGs suitable for implantation into any patient. Through utilizing the hiPSC platform, TEVGs for patients of vascular trauma can be developed on a massive scale, and of predictable and reproducible mechanical strength. Additionally, through developing and cryopreserving allogeneic universal hiPSC-ECs, these cells could be immediately applied to endothelialize TEVGs for small-diameter vascular intervention. Using hiPSC-VSMCs, we have successfully developed TEVGs with mechanical strength approaching that of saphenous veins, the common native grafts in vascular injury repair. Therefore, to achieve this future application of hiPSC-TEVGs, it is essential to establish the approach to efficiently decellularize the hiPSC-TEVGs, provide a luminal layer of hiPSC-ECs, and assess subsequent mechanical properties in vivo for preclinical efficacy as detailed in this proposal.

遭受过穿透性损伤（例如爆炸伤）或钝性损伤（例如骨折）造成的血管损伤的患者 和交通事故造成的挫伤）外伤通常会对其小直径外周造成严重损害 动脉（2-4mm，例如胫骨或肱动脉），需要手术干预以绕过或替换受伤的部位 血管段。由于血管外伤患者的隐静脉通达性可能较低 因爆炸伤或严重交通事故而导致一侧以上下肢严重受伤或丧失，以及 合成血管移植物的应用可能会因传染性病毒引起的潜在感染风险而受到阻碍 由于这些类型损伤的性质，迫切需要血管移植的替代来源。无细胞组织工程 因此，血管移植物（TEVG）可以为以下患者的紧急护理提供一种现成的治疗方法： 血管外伤。尽管具有显着机械强度的 TEVG 可以通过培养初级 人血管平滑肌细胞（VSMC）脱细胞化，这个平台有几个障碍 这阻止了它最终为血管创伤患者提供服务。有限的可扩展性、有限的可访问性、 原代 VSMC 之间的供体间功能差异可能会阻碍 TEVG 的生产效率。 此外，由于小直径 TEVG 在植入前需要自体内皮细胞 (EC) 涂层，以避免 血衣、患者 ECs 由于高龄或疾病而潜在的功能障碍，以及 获得和扩展患者自体 EC 所需的大量时间可能会妨碍其在治疗中的适用性 急性血管损伤。人类诱导多能干细胞 (hiPSC)，源自体细胞的自我更新细胞 细胞通过干细胞因子的异位表达，提供了解决并发症的绝佳替代方案 基于原代细胞的 TEVG。由于 hiPSC 几乎可以分化成任何体细胞类型，包括 VSMCs和ECs（hiPSC-VSMCs和hiPSC-ECs），这些细胞提供了无限的细胞来源以获得血管 细胞构建质量相当的 TEVG (hiPSC-TEVG)。此外，通过设计表达 可以建立人类白细胞抗原（HLA）蛋白、非或低免疫原性通用 hiPSC， 使 hiPSC-TEVG 适合植入任何患者体内。通过利用 hiPSC 平台，TEVG 针对血管创伤患者的治疗可以大规模开发，并且具有可预测性和可重复性 机械强度。此外，通过开发和冷冻保存同种异体通用 hiPSC-EC， 这些细胞可以立即应用于内皮化 TEVG，以进行小直径血管干预。使用 hiPSC-VSMCs，我们已成功开发出机械强度接近的TEVGs 大隐静脉是血管损伤修复中常见的天然移植物。因此，要实现这一未来的应用 对于 hiPSC-TEVG，有必要建立有效去细胞化 hiPSC-TEVG 的方法，提供 hiPSC-EC 的管腔层，并评估随后的体内机械特性以了解临床前功效 详细说明在本提案中。