Enhanced Propagation of Adult Human Renal Epithelial Cells

描述（由申请人提供）：现有的治疗肾衰竭的方法仍然是次优的，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，只替代滤过成分，而不考虑肾脏的稳态、调节、代谢和内分泌功能。为了满足对更先进治疗方法的需求，已经生产了含有猪肾上皮细胞（REC）的体外装置，并在败血症和ARF的临床前大型动物模型中被证明是有效的。一种含有人体细胞的中空纤维肾辅助装置（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，生物治疗肾装置正在开发中，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，可穿戴生物人工肾脏（WEBAK）正在为ESRD开发。这项创新技术从大型动物到临床环境的转变将需要功能性人类REC的产生，不仅要以适合设备制造的方式产生，而且要有足够的数量来治疗所有从肾脏替代疗法中受益的患者。仅ESRD目前就影响了超过43万美国患者，每年的成本超过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70万患者，目前有35万患者患有CRS。该研究计划涵盖了从人尸体肾脏中分离REC祖细胞群的增强方法，并将其繁殖到比当前方法大得多的生物量，然后最终分化为功能性REC。以这种方式获得的人REC将与传统分离的细胞在产量、体外形态和治疗潜力方面进行比较。每个供体的细胞产量将与预计需求相等，制造的技术可行性将通过使用冷冻保存的人类REC祖细胞构建WEBAK设备来清楚地证明。将强化繁殖技术成功应用于人类REC，将为制造数量显著增加的用于治疗ARF、ATN、MOF、败血症、CRS和ESRD的生物治疗装置提供生物质，这些疾病都有数十亿美元的市场，因此，将有助于挽救数千甚至数十万人的生命。

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.