Human Enterohepatic Cell Model for Predictive Toxicology

用于预测毒理学的人类肠肝细胞模型

• 批准号: 7012944
• 负责人: Jeffrey M. Macdonald
• 金额: $ 34.77万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7012944
• 关键词: NIH Roadmap Initiative tag; bioengineering /biomedical engineering; biomaterial development /preparation; biomimetics; bioreactors; cell cell interaction; cell line; drug metabolism; drug screening /evaluation; extracellular matrix; human tissue; intestinal mucosa; liver; liver cells; metabolomics; model design /development; nuclear magnetic resonance spectroscopy; pharmacokinetics; postmortem; tissue /cell culture; tissue engineering; total artificial organ; toxicology

DESCRIPTION (provided by applicant): It costs half a trillion dollars to bring a drug candidate to market. This expense is causing pharmaceutical companies to focus on compounds that have the greatest earning potential, orphan the development of critically needed drugs that have low revenue projections, and transfer high research costs to the consumer. All of these industry trends significantly increase the number of patients who can not access needed medicine because it is unavailable or unaffordable. The pharmaceutical industry is attempting to address this problem by heavily investing in human pre-clinical research as a mechanism to control cost and enhance the success of drug development. The industry is anxious for new tools to support the human pre-clinical research initiative. There is a particular need for advanced in vitro model systems to evaluate the toxicity of chemicals and drugs. This project's goal is to develop such a system. Abstract: The current in vitro technology for testing drug candidates is based on two-dimensional (2-D) sandwich livercell culturing techniques developed four decades ago. These In vivo models are complicated by the presence of structural and functional heterogeneity of biochemical pathways at the tissue and organism levels, and do not allow for mechanisms to be clearly defined or reproducibly examined. An in vitro system that is a much better model of a human liver is needed. Over the past dozen years this research team has been developing a state-of-the-art multicoaxial bioreactor (MCB) for creating the first human bioartificial liver. Over the past four years the team has focused on identifying the optimum human liver cell population for seeding the three-dimensional (3-D) bioreactor cultures. It has been determined that an unfractionated mixture of human liver cells shown to contain hepatic stem/progenitors provides favorable bioreactor results. In addition to this work the team has developed versatile NMR-compatible bioreactors that can obtain in situ metabolomics and fluxomics data. In this project we will create the first 3-D human bioartificial entero-hepatic organ-system and establish the feasibility of using this model system to evaluate the toxicity of chemicals and drugs. The proposed technology will be based on incorporating a defined population of human liver cells in an extracellular matrix thus creating a microenvironment composed of a precise composition of insoluble factors to interact with the cells. The encapsulated hepatocytes will be placed in a bioreactor compartment adjacent to a compartment containing a human derived- intestinal cell line, CaCo-2. This will be achieved using two multiple compartment bioreactor designs; a multicoaxial bioreactor (MCB) and a NMR-compatible bioreactor. The proposed bioreactor designs contain at least 4 compartments. This will permit the two cell types to interact across a small space and be perfused by separate plasma/intestinal compartments representing the blood and the intestinal compartments. This will mimic blood flow of the human entero-hepatic system. The advantage of the MCB bioreactor is that the intestinal barrier is better replicated. The advantage of the NMR-compatible bioreactor is that a novel interleaved NMR method can obtain in situ metabolomic and fluxomic data simultaneously from the two tissues. The expected result of this project will be an artificial human liver model that will be used with computational metabolomic and fluxomic analysis to identify toxicological and pharmacological drug targets.

描述（由申请人提供）： 将一种候选药物推向市场要花费5万亿美元。这一费用导致制药公司专注于具有最大盈利潜力的化合物，忽视了收入预测较低的急需药物的开发，并将高昂的研究成本转移给消费者。所有这些行业趋势都大大增加了无法获得所需药物的患者数量，因为药物不可用或负担不起。制药业正试图通过大量投资于人类临床前研究来解决这个问题，作为控制成本和提高药物开发成功率的机制。该行业迫切需要新的工具来支持人类临床前研究计划。特别需要先进的体外模型系统来评估化学品和药物的毒性。这个项目的目标就是开发这样一个系统。 摘要： 目前用于测试候选药物的体外技术是基于40年前开发的二维（2-D）夹心肝细胞培养技术。这些体内模型由于在组织和生物体水平存在生化途径的结构和功能异质性而变得复杂，并且不允许明确定义或可重复检查机制。需要一种更好的人体肝脏模型的体外系统。在过去的十几年里，该研究团队一直在开发最先进的多同轴生物反应器（MCB），以创建第一个人类生物人工肝。在过去的四年里，该团队一直致力于确定用于接种三维（3-D）生物反应器培养物的最佳人类肝细胞群。已经确定，显示含有肝干细胞/祖细胞的人肝细胞的未分级混合物提供有利的生物反应器结果。除了这项工作，该团队还开发了多功能NMR兼容的生物反应器，可以获得原位代谢组学和通量组学数据。 本计画将建立第一个三维人体生物人工肝肠器官系统，并探讨利用此模型系统评估化学药品毒性的可行性。所提出的技术将基于将确定的人类肝细胞群体并入细胞外基质中，从而创建由不溶性因子的精确组合物组成的微环境以与细胞相互作用。将包封的肝细胞置于与含有人源性肠细胞系CaCo-2的隔室相邻的生物反应器隔室中。这将使用两种多隔室生物反应器设计实现;多同轴生物反应器（MCB）和NMR兼容生物反应器。拟定的生物反应器设计包含至少4个隔室。这将允许两种细胞类型在小空间内相互作用，并由代表血液和肠隔室的单独血浆/肠隔室灌注。这将模拟人体肠-肝系统的血流。MCB生物反应器的优点是肠道屏障可以更好地复制。NMR兼容生物反应器的优点是，一种新的交错NMR方法可以同时从两种组织中获得原位代谢组学和通量组学数据。该项目的预期结果将是一个人工人肝模型，该模型将与计算代谢组学和通量组学分析一起使用，以确定毒理学和药理学药物靶点。