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: 7268038
• 负责人: Jeffrey M. Macdonald
• 金额: $ 33.76万
• 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/7268038
• 关键词: 3-Dimensional; Address; Adsorption; Alginates; Artificial Liver; Arts; Biochemical; Biochemical Pathway; Bioinformatics; Biological Models; Biomedical Engineering; Bioreactors; Blood; Blood flow; Cell Culture Techniques; Cell Extracts; Cell Line; Cell Therapy; Cell model; Cells; Chemicals; Collaborations; Collagen Type I; Compatible; Computer software; Condition; Cost Control; Culture Techniques; Cultured Cells; Data; Databases; Development; Drug Delivery Systems; Drug Industry; Encapsulated; Extracellular Matrix; Gases; Genotype; Glutamine; Glycine; Goals; Hepatic; Hepatocyte; Heterogeneity; Human; In Situ; In Vitro; Industry; Insulin; Intestines; Liver; Marketing; Medicine; Metabolic; Metabolic Pathway; Metabolism; Methods; Modeling; Monitor; Multinuclear NMR; Numbers; Organism; Orphan; Oxygen; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase III Clinical Trials; Phenobarbital; Phenotype; Plasma; Population; Pyruvate; Pyruvates; Rate; Rattus; Reaction; Recipe; Research; Resources; Stem cells; Stream; Sum; System; Technology; Therapeutic; Time; Tissues; Today; Toxic effect; Toxicology; Work; Writing; abstracting; analytical method; base; body system; cell preparation; cell type; cost; day; design; drug development; drug testing; in vitro Model; in vivo Model; mecarzole; metabolomics; novel; pre-clinical research; progenitor; stem; success; tool; trend; troglitazone; two-dimensional

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）： 将候选药物推向市场需要花费五万亿美元。这项费用导致制药公司专注于具有最大盈利潜力的化合物，放弃开发收入预测较低的急需药物，并将高昂的研究成本转嫁给消费者。所有这些行业趋势都显着增加了因无法获得或负担不起所需药物而无法获得所需药物的患者数量。制药行业正试图通过大力投资人类临床前研究来解决这个问题，以此作为控制成本和提高药物开发成功率的机制。业界迫切需要新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。该项目的目标是开发这样一个系统。 抽象的： 目前用于测试候选药物的体外技术基于四十年前开发的二维 (2-D) 夹心肝细胞培养技术。这些体内模型因组织和生物体水平上生化途径的结构和功能异质性的存在而变得复杂，并且不允许明确定义或可重复地检查机制。我们需要一种更好的人类肝脏模型体外系统。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），以创造第一个人类生物人工肝脏。在过去的四年里，该团队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，含有肝干/祖细胞的人肝细胞的未分级混合物提供了有利的生物反应器结果。除了这项工作之外，该团队还开发了多功能的 NMR 兼容生物反应器，可以获得原位代谢组学和通量组学数据。 在这个项目中，我们将创建第一个 3D 人体生物人工肠肝器官系统，并建立使用该模型系统评估化学品和药物毒性的可行性。所提出的技术将基于将特定的人类肝细胞群纳入细胞外基质中，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。封装的肝细胞将被放置在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。这将通过两个多隔室生物反应器设计来实现；多同轴生物反应器 (MCB) 和 NMR 兼容生物反应器。所提议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一个小空间内相互作用，并通过代表血液和肠室的单独血浆/肠室进行灌注。这将模拟人类肠肝系统的血流。 MCB生物反应器的优点是可以更好地复制肠道屏障。核磁共振兼容生物反应器的优点是，一种新颖的交错核磁共振方法可以同时从两种组织获得原位代谢组和通量组数据。该项目的预期结果将是一个人造人类肝脏模型，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。