New In Vitro Human Liver Toxicity Bioassay System

新型体外人肝毒性生物测定系统

基本信息

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
项目类别：
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

项目摘要

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和商业化人类3D或微型生物测定系统，适用于代谢功能和长期药物毒性概况的实时分析。目前，肝功能主要使用HEPG2细胞系，并在胶原蛋白或聚合物基质上培养的原代肝细胞研究。但是，由于代谢功能的快速丧失，这些2D单层模型仅允许部分或短期毒性分析。有趣的是，3D基质/支架培养物中的球体形成显示可改善宿主细胞功能。在第一阶段的研究中，我们成功地证明了新的人类生物遵循-3D Hubiogel（高密度凝胶）的可行性，以支持原代肝细胞的长期生存和维持。此外，旋转生物反应器系统的整合可以有效地生产/分析可行的微型肝球体。现在，我们建议完成新的3D肝生物测定平台的开发，该平台具有改善的代谢功能，因此将在体外进行精确的药物毒性测试。第二阶段的特定目标是：1。通过优化3D Hubiogel支架和旋转生物反应器或灌注培养策略来建立有效的肝细胞培养方案； 2。通过评估关键的生化和代谢终点来测试3D培养模型的测试功能；和3。通过对CYP450诱导和化学/药物毒性反应的实时分析来验证迷你肝脏测定系统的实际实用性。比较对照研究将包括I型胶原蛋白或Matrigel的3D培养支架以及新鲜的人类肝叶片培养物。 3D肝生物测定的高通量适应性也将通过CYP-微观阵列检查毒性组学。因此，将确认新的可销售3D肝生物测定系统的生物学相关性，以改善人类的药物毒性分析/预测。我们相信，这种先进的体外肝毒性模型将对基本，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：目前没有可接受的商业肝毒性测定法，它利用人类的生物体系培养系统。 VBI将开发适合HTS应用程序的市场就绪的3D肝测定平台。出售3D Hubiogel文化套件或内部生物测定服务将具有重要的全球市场，作为研究和诊断工具。