Enhanced Propagation of Adult Human Renal Epithelial Cells Allowing for Commercia

描述（由申请方提供）：肾细胞疗法有望改善急性肾衰竭（ARF）、急性肾小管坏死（ATN）、多器官衰竭（MOF）、脓毒症、心肾综合征（CRS）和终末期肾病（ESRD）患者的生存率和总体健康状况。为此，正在迅速开发解决肾脏替代疗法中被忽视的生理成分的治疗性生物装置。将这一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1阶段部分开发的增强繁殖（EP）技术从人类移植废弃物中获得。尽管人体组织的可用性仍然有限，但EP方案的开发极大地增强了肾祖细胞的扩增，消除了作为肾细胞治疗临床应用的限制因素的细胞可用性。本提案的总体目标是优化在生物人工肾上皮细胞系统（BRECS）中分离、繁殖、冷冻保存、分化、整合和维持人EP细胞所需的生产方案。接种EP人肾上皮细胞（HREC）的器械将用于证明在已建立的猪败血症模型中的有效性。将使用全血生物测定法在体外监测一组广泛的有效性标志物，并将结果与脓毒症猪模型中的治疗结果相关联。数据将用于制定根据EP方案分离和保存的细胞的有效性放行标准，并满足生物许可申请的效价要求。BRECS治疗的目标初始人群将是脓毒症诱导的全身炎症反应综合征（SIRS）的急性肾衰竭患者。将在该临床目标所需肾脏支持的预计7天临床时间过程中证明维持器械和体外回路的可行性。成功完成II期提案的数据将用于向美国食品药品监督管理局（FDA）提交研究性新药（IND）申请，以进行I/II期临床试验，评价接种了来自增强繁殖方案的HREC的生物可降解材料器械的安全性和有效性。 公共卫生相关性：本研究的长期目标是开发和优化用于分离、扩增和维持人肾上皮细胞（HREC）的增强繁殖（EP）方案的生成和鉴定，以用于生物学应用，从而消除生成HREC的有限组织/细胞来源的问题。本研究提案中针对的具体应用是评估EP HREC在生物人工肾上皮细胞系统（BRECS）中的利用，该系统有效地为各种疾病过程增加了治疗价值，包括全身炎症反应综合征（SIRS）和脓毒症的治疗。在美国，每年有200，000名患者发生严重脓毒症伴SIRS，即使使用重症监护室和广谱抗生素，死亡率也为30- 40%。成功完成本提案中计划的研究将允许在BRECS应用中使用无限的细胞源。该装置将大大减少脓毒症和SIRS的多器官效应，从而改善受这些疾病过程影响的患者的临床结果。

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.