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: 10439796
• 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/10439796
• 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.

因穿透性（如爆炸伤）或钝性（如骨折）导致血管创伤的患者 和交通事故造成的挫伤）创伤经常会对他们的小直径外周造成严重损伤 动脉（2- 4 mm，例如胫动脉或肱动脉），需要手术干预以绕过或替换受伤的 血管节段由于大隐静脉的可及性在血管创伤患者中可能较低， 由于爆炸伤或严重交通事故造成的严重伤害或一个以上下肢的丧失，以及 合成血管移植物的应用可能受到潜在的感染风险的阻碍 由于这些类型损伤的性质，迫切需要血管移植物的替代来源。脱细胞组织工程 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： 血管创伤尽管具有显著机械强度的TEVG可以从培养原代细胞中开发出来， 人血管平滑肌细胞（VSMC）随后去细胞化，该平台存在几个障碍 这就阻止了它最终为血管创伤的受害者服务。有限的可扩展性，有限的可访问性， 并且原代VSMC中供体与供体的功能差异可能阻碍TEVG生产的效率。 此外，由于小直径的TEVG在植入之前需要自体内皮细胞（EC）涂层，以避免 血衣，由于高龄或疾病导致的患者EC的潜在功能障碍，以及 获得和扩增患者的自体EC需要大量时间，这可能妨碍治疗的适用性。 急性血管损伤人诱导多能干细胞（hiPSC），来源于体细胞的可自我再生细胞， 细胞通过干细胞因子的异位表达，为解决并发症提供了一个很好的替代方案 基于原电池的TEVG。由于hiPSC可以分化成几乎任何体细胞类型，包括 VSMC和EC（hiPSC-VSMC和hiPSC-EC），这些细胞提供了无限的细胞来源以获得血管内皮细胞。 细胞以构建质量相当的TEVG（hiPSC-TEVG）。此外，通过工程化表达 人白细胞抗原（HLA）蛋白，可以建立非免疫原性或低免疫原性的通用hiPSC， 使hiPSC-TEVG适合植入任何患者。通过利用hiPSC平台，TEVG 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，用于小直径血管介入。使用 hiPSC-VSMCs，我们已经成功地开发了TEVG，其机械强度接近 隐静脉是血管损伤修复中常见的自体移植物。因此，要实现这一未来的应用 因此，必须建立有效地使hiPSC-TEVG脱细胞的方法， hiPSC-EC的管腔层，并评估随后的体内机械特性以获得临床前疗效， 在这个提案中详细说明。