Endothelial cell transplantation for multi-organ repair to counter radiation injury

摘要 暴露在电离辐射中通常是致命的，因为急性辐射综合征(ARS)表现为 胃肠-ARS(GI-ARS)和造血-ARS(H-ARS)。急性辐射的延迟效应 暴露(DEARE)会导致多器官功能障碍综合征(MODS)。的共同点 辐射引起的多器官衰竭是由于内皮细胞和淋巴管内皮细胞的损伤， 导致渗漏、凝血障碍和炎症，为感染、硬化症和 肿瘤发生学。辐射诱导内皮细胞功能障碍的分子基础尚不清楚。我们的目标 是通过静脉移植容易获得的、不需要的血管内皮细胞来利用其再生功能。 货架，同种异体人类内皮细胞，以减轻ARS，DEARE和MODS。我们的中心假设是辐射 受损的血管和淋巴管内皮细胞功能障碍，无法执行其血管 功能或提供促进组织愈合所需的指导性信号，从而导致ARS和 迪尔。我们认为，照射后一天或几天移植正常的促再生内皮细胞可以 抢救辐射损伤内皮细胞多器官缺损区，促进无疤痕愈合。我们已经展示了 组织特异性内皮细胞通过产生血管分泌生长因子来协调受损器官的修复 没有纤维化。人内皮细胞静脉移植亚致死性造血恢复 经辐射的啮齿动物和经致死辐射的非人类灵长类(NHP) 肿瘤发生学。提议的实验的基本原理是，如果我们知道如何有效地生成 大量现成的GMP级人脐静脉内皮细胞(HUVECs)作为“同种异体血管” ，我们将使用NHP大动物模型辐射模型来确定HUVEC的药代动力学 移植，将其作为支持器官的最终或中间放射对策 辐射后修复。我们将通过解决这些目标来检验这一假设：1)制造丰富的 猴内皮细胞(MUVECs)和人内皮细胞(HUVECs)的功能性临床级主细胞库 静脉移植。2)确定同种/异种MUVEC的关键参数和 人脐静脉内皮细胞移植到受体小鼠体内减轻照射后H-ARS和GI-ARS损伤 引发纤维化..3)使用辫尾猕猴NHP辐射模型来确定计划、安全性 移植微血管内皮细胞和人脐静脉内皮细胞修复多器官损伤的效果 不留疤痕。拟议研究的完成将使同种异体现成的治疗用途成为可能 “人类内皮细胞”暂时回到受损的血管床上，这些血管床被辐射器官修复 血管生成和血管壁龛功能促进器官修复、疤痕形成。这些研究的成功 将为急慢性疾病的治疗提供现成的医疗对策(MCM) 预防危及生命的并发症的辐射综合征。 1

Abstract Exposure to ionizing radiation is often fatal due to acute radiation syndromes (ARS) manifested as Gastrointestinal-ARS (GI-ARS) and Hematopoietic-ARS (H-ARS). Delayed effects of acute radiation exposure (DEARE) lead to multi-organ dysfunction syndrome (MODS). A common denominator of radiation induced multi-organ failure is due to damage to endothelial cells (ECs) and lymphatic ECs, resulting in leakiness, coagulopathy and inflammation, setting up stage for infection, sclerosis and tumorigenesis. The molecular basis of radiation-induced EC dysfunction is not well understood. Our goal is to capitalize on the regenerative function of ECs by intravenously transplanting readily-available, off-the- shelf, allogeneic human ECs to mitigate ARS, DEARE and MODS. Our central hypothesis is that radiation damaged blood vessel and lymphatic ECs become dysfunctional and fail to perform their vascular functions or supply the instructive signals required to promote tissue healing thereby leading to ARS and DEARE. We propose that transplantation of normal pro-regenerative ECs a day or days after radiation can rescue the multi-organ defects of radiation-injured ECs and promote scar-free healing. We have shown that tissue-specific ECs by producing angiocrine growth factors orchestrate the repair of injured organs without fibrosis. Intravenous transplantation of human ECs restores hematopoietic recovery in sublethally irradiated rodents and lethally irradiated pigtail macaque non-human primates (NHP) without fibrosis or tumorigenesis. The Rationale for the proposed experiments is that if we know how to efficiently generate abundant off-the-shelf GMP-grade human umbilical vein ECs (HUVECs) as a “generic allogeneic vascular graft”, we will use NHP large animal model radiation models to determine the pharmacokinetics of HUVEC transplantation to use them as a definitive or intermediary radiation countermeasure to support organ repair post-radiation. We will test this hypothesis by addressing these Aims: 1) Manufacture of abundant functional clinical-grade master cell banks of monkey ECs (MUVECs) and human (HUVECs) for intravenous transplantation. 2) Identify the critical parameters for allogeneic/xenogeneic MUVEC and HUVEC transplantation into recipient mice to mitigate post-irradiation H-ARS and GI-ARS injury without provoking fibrosis..3) Employ pigtail macaque NHP radiation models to determine the scheduling, safety and efficacy of transplanting MUVECs and HUVECs to rejuvenate vascular niche for multi-organ repair without scarring. Completion of the proposed studies will enable therapeutic use of allogeneic off-the-shelf “human ECs” that transiently home to the disrupted vascular beds of irradiated organs restoring angiogenesis and vascular niche functions promoting organ repair, scarring. The success of these studies will provide for a readily available medical counter measure (MCM) for the treatment of acute and chronic radiation syndromes preventing life threatening complications. 1