Enhanced Propagation of Adult Human Renal Epithelial Cells

描述（由申请方提供）：可用的肾衰竭治疗仍然是次优的，急性肾衰竭（ARF）的相关死亡率超过50%，终末期肾病（ESRD）的预期寿命仅为4 - 5年。目前的肾脏替代疗法是非生理性的，仅替代过滤成分，而不考虑肾脏的稳态、调节、代谢和内分泌功能。为了满足对更先进疗法的需求，已经生产了含有猪肾上皮细胞（REC）的体外装置，并证明其在败血症和ARF的临床前大型动物模型中有效。含有人类细胞的基于中空纤维的肾辅助装置（RAD）已经在FDA批准的I/II期和II期临床试验中进行了评估。目前，双肾装置正在开发中，以改善ARF、急性肾小管坏死（ATN）、多器官衰竭（MOF）、败血症和心肾综合征（CRS）的影响。更具体地说，正在为ESRD开发可穿戴生物人工肾（WEBAK）。这项创新技术从大型动物向临床环境的过渡将要求功能性人类REC不仅以适合器械制造的方式产生，而且以足够的数量产生，以治疗从肾脏替代治疗中受益的所有患者人群。目前，仅ESRD就影响了超过430，000名美国患者，每年花费超过250亿美元，预计到2030年ESRD患者人数将增加到224万。脓毒症导致ARF、ATN或MOF，每年影响700，000名患者，目前有350，000名患者患有CRS。本研究建议包括增强的方法分离REC祖细胞群体从人尸体肾脏和繁殖到一个更大的生物量比目前的协议，然后通过终端分化为功能REC。人类REC以这种方式获得将比较传统的分离细胞的产量，体外形态和治疗潜力。每个供体的细胞产量将等同于预计的需求，生产的技术可行性将通过使用冻存的人REC祖细胞构建WEBAK装置来明确证明。成功地将增强的繁殖技术应用于人类REC将为制造用于治疗ARF、ATN、MOF、败血症、CRS和ESRD的显著更大数量的生物制品装置提供生物质，所有疾病都具有数十亿美元的市场，因此，将有助于挽救数千人（如果不是数十万人）的生命。 公共卫生相关性 肾细胞治疗向临床环境的过渡将需要制造含有人源细胞的生物工程装置。目前，人体组织的可用性有限。因此，必须优化来自可用的人肾移植废弃物的肾上皮细胞的扩增和表征，并且在是否可以制造足够数量的装置以有效治疗迅速增加的患有慢性和急性肾脏疾病的患者的背景下评估治疗潜力。

DESCRIPTION (provided by applicant): Available therapies for renal failure continue to be suboptimal with associated mortality rates of over 50% for acute renal failure (ARF) and a life expectancy for end stage renal disease (ESRD) of only 4-5 years. Current renal replacement therapies are non-physiologic, replacing only the filtrative component with no regard to the homeostatic, regulatory, metabolic and endocrine functions of the kidney. To address the need for more advanced therapies, extracorporeal devices containing porcine renal epithelial cells (REC) have been produced and proven to be efficacious in pre-clinical large animal models of sepsis and ARF. A hollow fiber based renal assist device (RAD) containing human cells has been evaluated in FDA-approved Phase I/II and Phase II clinical trials. Currently, biotherapeutic renal devices are in development to ameliorate the effects of ARF, acute tubular necrosis (ATN), multiple organ failure (MOF), sepsis and cardiorenal syndrome (CRS). More specifically, wearable bioartificial kidneys (WEBAK) are being developed for ESRD. The transition of this innovative technology from large animals to the clinical setting will require functional human REC to be generated not only in a manner suitable for device manufacture but in sufficient quantities to treat all patient populations who stand to benefit from renal replacement therapy. ESRD alone currently effects over 430,000 U.S. patients and has an annual cost of more than 25 billion dollars with ESRD patient numbers expected to increase to 2.24 million by 2030. Sepsis leads to ARF, ATN or MOF affecting 700,000 patients annually and currently 350,000 patients suffer from CRS. This research proposal covers enhanced methods for the isolation of REC progenitor cell populations from human cadaver kidneys and propagation to a much larger biomass than current protocols, followed by terminal differentiation to functional REC. Human REC obtained in this manner will be compared to traditionally isolated cells for yield, in-vitro morphology and therapeutic potential. Cell yield per donor will be equated with projected need and technical feasibility of manufacture will be clearly demonstrated by the construction of a WEBAK device using cryopreserved human REC progenitors. Successful application of enhanced propagation techniques to human REC will provide the biomass for the manufacture of a significantly larger number of biotherapeutic devices for treatment of ARF, ATN, MOF, sepsis, CRS and ESRD, all diseases with multibillion dollar markets and, as result, will help save thousands, if not hundreds of thousands, of lives. PUBLIC HEALTH RELEVANCE Transition of renal cell therapy to the clinical setting will require the manufacture of bioengineered devices containing cells of human origin. Currently, the availability of human tissue is limited. Therefore, the amplification and characterization of renal epithelial cells from available human kidney transplant discards must be optimized and therapeutic potential evaluated in the context of whether or not devices can be made in sufficient numbers to effectively treat the burgeoning number of patients suffering from both chronic and acute renal diseases.