Enhanced Propagation of Adult Human Renal Epithelial Cells Allowing for Commercia

描述(申请人提供)：肾细胞疗法有望改善患有急性肾功能衰竭(ARF)、急性肾小管坏死(ATN)、多器官衰竭(MOF)、败血症、心肾综合征(CRS)和终末期肾病(ESRD)的患者的存活率和整体健康。为此，针对肾脏替代疗法中被忽视的生理成分的治疗性生物装置正在迅速发展。将这项创新技术转移到临床环境将需要使用大量人类来源的细胞制造设备。细胞将使用本提案第一阶段中开发的增强繁殖(EP)技术从人类移植废物中提取。尽管人体组织的可获得性仍然有限，但EP方案的发展极大地增强了肾细胞前体细胞的扩增，消除了细胞可获得性这一限制因素，使肾脏细胞治疗的临床应用受到限制。这项建议的总体目标是优化在生物人工肾上皮细胞系统(BRECS)中分离、增殖、冷冻保存、分化、整合和维护人EP细胞所需的方案。种植EP人肾上皮细胞(HRECs)的装置将用于证明在已建立的猪脓毒症模型中的疗效。在败血症猪模型中，将使用全血生物测定和与治疗结果相关的结果在体外监测广泛的疗效标记物。数据将被用来为EP协议下分离和维护的细胞制定与有效性相关的释放标准，并满足生物许可证申请的效力要求。BRECS治疗的初始目标人群将是合并脓毒症引起的全身炎症反应综合征(SIRS)的急性肾功能衰竭患者。维持该装置和体外循环的可行性将在这一临床目标所需的肾脏支持的预计7天临床时间过程中得到证明。从成功完成第二阶段提案获得的数据将用于向食品和药物管理局(FDA)提交一份研究新药(IND)，以进行I/II阶段临床试验，评估从增强型繁殖方案中获得的以HREC为种子的生物治疗设备的安全性和有效性。 公共卫生相关性：这项研究的长期目标是开发和优化用于生物应用的人肾上皮细胞(HREC)分离、扩增和维护的增强型繁殖(EP)方案的生成和鉴定，从而消除用于生成HREC的有限组织/细胞来源的问题。这项研究建议的具体应用是评估EP HRECs在生物人工肾上皮细胞系统(BRECS)中的使用情况，该系统可有效地增加各种疾病过程的治疗价值，包括治疗全身炎症反应综合征(SIRS)和脓毒症。在美国，每年有20万名患者发生严重的全身炎症反应综合征，即使使用重症监护病房和广谱抗生素，死亡率也高达30%-40%。成功完成本提案中计划的研究将允许在金砖四国的应用中使用无限的细胞来源。该设备将极大地减少脓毒症和全身炎症反应综合征的多器官影响，从而改善受这些疾病过程影响的患者的临床结局。

DESCRIPTION (provided by applicant): Renal cell therapy promises to improve the survival and overall health of patients suffering from acute renal failure (ARF), acute tubular necrosis (ATN), multiple organ failure (MOF), sepsis, cardiorenal syndrome (CRS) and end stage renal disease (ESRD). To this end, therapeutic biological devices addressing the neglected physiologic component of renal replacement therapy are rapidly being developed. Transfer of this innovative technology to the clinical setting will require the manufacture of devices using a large number of cells of human origin. Cells will be derived from human transplant discards using enhanced propagation (EP) techniques developed in the Phase 1 component of this proposal. Although the availability of human tissue remains limited, development of the EP protocol has greatly enhanced the amplification of renal cell progenitors, removing cell availability as the limiting factor to the clinical application of renal cell therapy. The overall goal of this proposal is to optimize the protocols necessary for the isolation, propagation, cryopreservation, differentiation, integration and maintenance of human EP cells in a bioartificial renal epithelial cell system (BRECS) with regard to manufacturing. Devices seeded with EP human renal epithelial cells (HRECs) will be used to demonstrate efficacy in a well established porcine model of sepsis. A broad panel of efficacy markers will be monitored in vitro using a whole blood bioassay and results correlated with therapy outcome in the septic pig model. Data will be used to develop release criteria for cells isolated and maintained under EP protocols relative to efficacy and to satisfy the potency requirement of the Biologic License Application. The targeted initial population for BRECS therapy will be acute renal failure patients with sepsis induced Systemic Inflammatory Response Syndrome (SIRS). The feasibility of maintaining the device and extracorporeal circuit will be demonstrated over the projected 7 day clinical time course of required renal support for this clinical target. Data derived from the successful completion of the Phase II proposal will be used for an Investigational New Drug (IND) submission to the Food and Drug Administration (FDA) for a Phase I/II clinical trial evaluating the safety and efficacy of biotherapeutic devices seeded with HREC derived from enhanced propagation protocols. PUBLIC HEALTH RELEVANCE: The long-term goal of this research is to develop and optimize the generation and qualification of enhanced propagation (EP) protocols for the isolation, expansion and maintenance of human renal epithelial cells (HREC) for use in biologic applications, thus eliminating the issue of limited tissue/cell sources for generation of HRECs. The specific application targeted in this research proposal is for the assessment of utilization of EP HRECs in a bioartificial renal epithelial cell system (BRECS) that effectively adds therapeutic value to a variety of disease processes, including the treatment of Systemic Inflammatory Response Syndrome (SIRS) and sepsis. Severe sepsis with SIRS occurs in 200,000 patients annually in the U.S. and has a mortality rate of 30-40%, even with use of intensive care units and broad spectrum antibiotics. Successful completion of the planned studies in this proposal will allow for an unlimited cell source for use in the BRECS application. This device would greatly reduce the multiorgan effects of sepsis and SIRS, thus improving the clinical outcome of patients affected by these disease processes.