Enhanced production of human hepatocytes from livers declined for transplant

肝脏产生的人类肝细胞产量因移植而下降

基本信息

批准号：
9140604
负责人：
MARK G CLEMENS
金额：
$ 36.61万
依托单位：
HEPATOSYS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-17 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9140604
关键词：
Address Animal Model Biomedical Engineering Cardiac Death Cell Line Cells Clinical Trials Collaborations Coupled Cryopreservation Development Drug Approval Failure Goals Gold Health Health Care Costs Hepatocyte Hepatotoxicity Hour Human Incubated Investments Ischemia Left Legal patent Liver Marketing Membrane Metabolism Methods Modification Organ Organ Preservation Outcome Perfusion Pharmaceutical Preparations Phase Preclinical Drug Evaluation Procedures Process Production Rattus Research Research Personnel Resuscitation Series Site Source System Technology Testing Time Toxicity Tests Translating Transplantation Warm Ischemia Xenobiotics base commercialization cost design drug development drug testing efficacy testing experience flexibility functional restoration improved improved functioning innovation innovative technologies liver injury novel therapeutics oxidation pre-clinical preclinical study screening species difference

项目摘要

DESCRIPTION (provided by applicant): Liver toxicity is a major barrier to the FDA approval of many new drugs. Moreover, since the liver is the major site of metabolism of most xenobiotics, it is essential to know how the liver metabolizes a potential new drug. Because of the many species differences between humans and commonly used animal models, testing on human hepatocytes remains the gold standard for drug screening. However, because the source of these hepatocytes is usually from livers that have been declined for use in transplantation, typically because of extended ischemic time, the supply of high quality cells is severely limited. The long-term goal of this project is to reliably improve the availability and viability of primary human hepatocytes isolated from non-transplantable Donation after Cardiac Death (DCD) livers through the modification of a hypothermic machine perfusion (HMP) resuscitation system that we have developed to restore function to ischemia damaged DCD livers for use in transplantation. Human hepatocytes are typically isolated from the pool of non-transplantable DCD organs which has been estimated to be as high as 40% of the current donor pool. However, inconsistent yield and quality of the isolated hepatocytes from these donors has led to limited availability resulting in higher costs for these cells. Major factors contributing to this inconsistency are the extended periods of warm ischemia experienced by these organs and the preservation process (simple cold storage) which does not resuscitate these organs. The HepatoSys solution (HS) coupled with HMP was developed to restore depleted energy stores, maintain membrane stability, and minimize oxidation damage resulting in a resuscitated liver. Our results indicate that our process can be used to successfully isolate high quality hepatocytes from otherwise unusable livers. In addition, we will add further commercial value to the technology by testing prolonged HMP of livers to provide greater flexibility in making fresh cells available, and the ability of additional oxygenated cold storage of hepatocytes in the HS to improve the quality of freshly isolated and cryopreserved hepatocytes. To achieve these goals, we propose the following aims. Specific Aim 1: Test the efficacy of prolonged HMP of livers with the HepatoSys solution on yield, viability, plateability and function of fresh hepatocytes isolated from rat DCD livers. Test the effect of up to 48 hours HMP followed by hepatocyte isolation on yield, viability, plateability and function of freshly plated isolated hepatocytes from DCD rats. Specific Aim 2: Test the efficacy of additional oxygenated cold storage (CS) of fresh hepatocytes after isolation in the HepatoSys solution on yield, viability, plateability and functio from DCD rat livers Test the ability of additional CS of isolated hepatocytes in the HS from DCD rats to maintain or improve the viability, plateability of freshly isolated rat hepatocytes. Specifc Aim 3: Test the efficacy of optimized conditions identified in aims 1 and 2 on yield, viability, plateability and function of freshly isolated hepatocytes from a small group of human DCD livers. Completion of these specific aims will provide a proof of concept that this technology has a potentially commercially valuable method to obtain viable, plateable and functional hepatocytes from livers currently not usable while providing greater flexibility in making freshly isolated hepatocytes available which will allow TRL and HepatoSys to increase market share. A Phase II application will seek to add further commercial value to the technology by expanding the pool to even poorer quality donor livers and improving the quality of cryopreserved hepatocytes.

描述（由适用提供）：肝毒性是FDA批准许多新药的主要障碍。此外，由于肝脏是大多数异种生物的代谢的主要部位，因此必须知道肝脏如何代谢潜在的新药。由于人类与常用动物模型之间存在许多物种差异，因此对人肝细胞的测试仍然是药物筛查的金标准。但是，由于这些肝细胞的来源通常来自肝脏被降低用于移植的肝脏，通常是由于延长的缺血时间，因此高质量细胞的供应受到严重限制。该项目的长期目标是可靠地提高主要的可用性和生存能力心脏死亡后的不可移植捐赠分离的人肝细胞（DCD）通过修改低温机器灌注（HMP）复苏系统而生存，我们已经开发出来恢复缺血功能损害DCD的功能，以损害DCD的生命，以便在移植中使用。人肝细胞通常是从不可移植的DCD器官库中分离出来的，据估计，该器官估计高达当前供体池的40％。但是，这些供体的孤立肝细胞的产量和质量不一致导致可用性有限，从而导致这些细胞的成本更高。导致这种不一致的主要因素是这些器官经历的温暖缺血的延长和保存过程（简单的冷藏）不会恢复这些器官。开发了与HMP结合的HEPATOSYS溶液（HS），以恢复已部署的能量储存，保持膜稳定性并最大程度地减少氧化损伤，从而导致肝脏复苏。我们的结果表明，我们的过程可用于成功地将高质量的肝细胞与原本无法使用的生活隔离。此外，我们将通过测试延长寿命的延长HMP来为该技术增加进一步的商业价值，从而提供更大的灵活性，以使新鲜细胞可用，以及HS中额外的氧化肝细胞的额外氧合冷储存的能力，以提高新鲜分离和冷冻保存的肝细胞的质量。为了实现这些目标，我们提出以下目标。特定目标1：通过hepatosys解决方案测试新鲜肝细胞的产量，生存力，平稳性和功能，测试寿命延长寿命的效率从老鼠DCD生命。测试高达48小时HMP的影响，然后测试肝细胞分离的效果，生存能力，平稳性和功能，使新鲜镀的DCD大鼠分离的分离的肝细胞的作用。具体目的2：测试新鲜肝细胞的额外氧化冷储存（Cs）在肝脏溶液中分离后的产量，可行性，平稳性和功能性的DCD大鼠生命测试的能力，可从DCD大鼠中维持或提高新鲜型号的HS中分离的肝细胞的其他CS的能力。特定目标3：测试目标1和2中确定的优化条件的效率，其产量，生存能力，平稳性以及来自一小群人DCD客人的新鲜分离肝细胞的功能。这些特定目标的完成将提供概念证明，该技术具有一种潜在的商业价值的方法，可以从目前无法使用的肝脏获得可行，可静止和功能的肝细胞，同时提供更大的灵活性，可以使新鲜隔离的肝细胞可用，从而可以允许TRL和Hepatosys增加市场份额。第二阶段的应用程序将寻求通过将池扩展到较差的质量供体生活并提高冷冻保存的肝细胞的质量来提高该技术的进一步商业价值。