New In Vitro Human Liver Toxicity Bioassay System

新型体外人肝毒性生物测定系统

• 批准号: 8458114
• 负责人: RAJ K. SINGH
• 金额: $ 29.72万
• 依托单位: VIVO BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-19 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8458114
• 关键词: 3-Dimensional; Acute; Adherent Culture; Agreement; Antineoplastic Agents; Area; Basic Science; Biochemical; Biological; Biological Assay; Biomedical Research; Bioreactors; Businesses; Cell Line; Cell Survival; Cell physiology; Cells; Chemicals; Chronic; Clinical; Collagen; Collagen Type I; Controlled Study; Culture Techniques; Development; Diagnostics Research; Dose; Drug Monitoring; Drug toxicity; Enzyme-Linked Immunosorbent Assay; Exhibits; Future; Gel; Goals; Growth; Hepatic; Hepatocyte; Hepatotoxicity; Housing; Human; In Vitro; Joint Ventures; Legal patent; Liver; Maintenance; Marketing; Metabolic; Metabolism; Methods; Microscopy; Microspheres; Modeling; Molecular Analysis; Monitor; Outcome; Perfusion; Pharmacologic Substance; Pharmacotherapy; Phase; Physiological; Polymers; Production; Progress Reports; Protocols documentation; Reaction Time; Relative (related person); Reporting; Reproducibility; Research; Reverse Transcriptase Polymerase Chain Reaction; Sales; Series; Services; Slice; Small Business Innovation Research Grant; Study models; System; Technology; Testing; Time; Tissue Sample; Toxic effect; Toxicity Tests; Toxicogenomics; base; cell type; comparative; density; design; drug metabolism; enzyme activity; fluorescence imaging; gene induction; human tissue; improved; indexing; liver function; matrigel; monolayer; novel; phase 1 study; pre-clinical research; research and development; response; scaffold; screening; tool; toxicant; tumor; two-dimensional

DESCRIPTION (provided by applicant): The overall objective of this SBIR proposal is to develop and commercialize a human 3D or mini- liver bioassay system suitable for real-time analysis of metabolic functions and long-term drug toxicity profiles. At present, liver functions are mostly studied using HepG2 cell line and primary hepatocytes cultured onto collagen or polymer matrices. However, these 2D monolayer models allow only partial or short-term toxicity analysis due to rapid loss of metabolic functions. Interestingly, spheroid formation in 3D matrix/scaffold cultures is shown to improve host cell functions. In Phase I studies, we successfully demonstrated the feasibility of a new human bioscaffold- 3D HuBiogel (high-density gel) for supporting long-term survival and maintenance of primary liver cells. Moreover, integration of rotary bioreactor system allowed efficient production/analysis of viable mini-liver spheroids. We now propose to complete the development of new 3D-liver bioassay platform that exhibits improved metabolic functions, and thus will enable precise drug toxicity testing in vitro. Phase II specific goals are: 1. Establish an efficient hepatic cells cultivation protocol by optimizing 3D HuBiogel scaffolds and rotary bioreactor or perfusion culture strategies; 2. Test functionality of 3D-liver culture model by evaluating key biochemical and metabolism endpoints; and 3. Validate the practical utility of mini-liver assay system via real- time analysis of CYP450 induction and chemical/drug toxicity responses. Comparative control studies will include 3D culture scaffolds of Type-I collagen or Matrigel as well as fresh human liver-slice cultures. High throughput adaptability of 3D-liver bioassay will also be examined for toxicogenomics via CYP-microarrays. Thus, biologic relevance of new marketable 3D-liver bioassay system will be confirmed for improved drug toxicity analysis/prediction in humans. We are confident this advanced in vitro hepatotoxicity model will positively impact basic, preclinical and biomedical research arenas. Commercial Importance & Significance: No acceptable commercial liver toxicity assay currently exists which utilizes a human bio-scaffold culture system. VBI will develop a market-ready 3D-liver assay platform adaptable to HTS applications. Sale of 3D HuBiogel culture kits or in-house bioassay services would have significant world-wide market, as a research & diagnostic tool.

描述（由申请人提供）：该 SBIR 提案的总体目标是开发并商业化适用于实时分析代谢功能和长期药物毒性特征的人体 3D 或微型肝脏生物测定系统。目前，肝功能主要使用在胶原或聚合物基质上培养的 HepG2 细胞系和原代肝细胞来研究。然而，由于代谢功能的快速丧失，这些二维单层模型仅允许部分或短期毒性分析。有趣的是，3D 基质/支架培养物中球体的形成被证明可以改善宿主细胞功能。在第一阶段研究中，我们成功证明了新型人体生物支架——3D HuBiogel（高密度凝胶）支持原代肝细胞长期存活和维持的可行性。此外，旋转生物反应器系统的集成可以有效生产/分析可行的微型肝球体。我们现在建议完成新的 3D 肝脏生物测定平台的开发，该平台具有改善的代谢功能，从而能够实现精确的体外药物毒性测试。 II期具体目标是： 1.通过优化3D HuBiogel支架和旋转生物反应器或灌注培养策略，建立高效的肝细胞培养方案； 2. 通过评估关键生化和代谢终点来测试3D肝脏培养模型的功能； 3. 通过实时分析 CYP450 诱导和化学/药物毒性反应，验证微型肝脏测定系统的实用性。比较对照研究将包括 I 型胶原蛋白或基质胶的 3D 培养支架以及新鲜的人类肝脏切片培养物。 3D 肝脏生物测定的高通量适应性也将通过 CYP 微阵列进行毒物基因组学检查。因此，新的市售 3D 肝脏生物测定系统的生物学相关性将得到证实，以改善人体药物毒性分析/预测。我们相信这种先进的体外肝毒性模型将对基础、临床前和生物医学研究领域产生积极影响。商业重要性和意义：目前尚不存在利用人体生物支架培养系统的可接受的商业肝毒性测定。 VBI 将开发一个适合市场的 3D 肝脏检测平台，适用于 HTS 应用。作为一种研究和诊断工具，3D HuBiogel 培养套件或内部生物测定服务的销售将拥有重要的全球市场。