Enhanced Propagation of Adult Human Renal Epithelial Cells

描述(由申请人提供)：现有的治疗肾功能衰竭的方法仍然不理想，急性肾功能衰竭(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学的，只替换滤过性成分，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，含有猪肾上皮细胞(REC)的体外装置已经被生产出来，并被证明在临床前的大动物脓毒症和ARF模型中是有效的。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，用于改善ARF、急性肾小管坏死(ATN)、多器官功能衰竭(MOF)、脓毒症和心肾综合征(CRS)的生物治疗性肾脏装置正在开发中。更具体地说，正在为终末期肾病开发可穿戴生物人工肾脏(WEBAK)。这项创新技术从大型动物到临床环境的过渡将要求不仅以适合设备制造的方式产生功能性人类REC，而且需要产生足够数量的REC来治疗所有将从肾脏替代疗法中受益的患者群体。仅ESRD一项目前就影响了超过43万名美国患者，每年的成本超过250亿美元，预计到2030年，ESRD患者数量将增加到224万人。脓毒症导致ARF、ATN或MOF，每年影响70万名患者，目前有35万名患者患有CRS。这项研究建议包括改进的方法，从人身体肾脏分离REC祖细胞群体，并繁殖到比当前方案大得多的生物量，然后向功能性REC终末分化。以这种方式获得的人REC将在产量、体外形态和治疗潜力方面与传统分离的细胞进行比较。每个捐赠者的细胞产量将等同于预计的需求，制造的技术可行性将通过使用冷冻保存的人REC祖细胞构建WEBAK设备而清楚地得到证明。将增强型繁殖技术成功应用于人类REC，将为生产更多用于治疗ARF、ATN、MOF、脓毒症、CRS和ESRD的生物治疗设备提供生物量，所有这些疾病都具有数十亿美元的市场，因此将有助于拯救数千人(如果不是数十万人)的生命。 公共卫生相关性 将肾脏细胞治疗过渡到临床环境将需要制造包含人类来源细胞的生物工程设备。目前，人体组织的可获得性有限。因此，必须从现有的人类肾脏移植废物中优化肾上皮细胞的扩增和鉴定，并根据是否可以制造足够数量的设备来有效治疗迅速增长的慢性和急性肾脏疾病患者来评估治疗潜力。

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.