Micro-Liver Platform Development for Evaluating Drug Disposition and Toxicity In

描述（由申请方提供）：肝毒性是药物化合物（即Rezulin、Prexige）上市前和上市后损耗的主要原因。器官功能的显著物种特异性差异现在需要用旨在评估人类对药物反应的测定来补充动物数据。原代人肝细胞贴壁培养被认为是评价临床前药物代谢、酶诱导和肝脏特异性毒性的金标准。然而，在常规用于药物开发的常规培养条件下，肝细胞显示肝脏特异性功能快速（小时）下降。最近，已经开发了一种稳健的人肝组织模型，其具有优化的微尺度结构，采用行业标准的多孔格式，在体外保留肝脏特异性功能4-6周。该微型平台中的肝细胞分泌肝脏特异性产物，展示功能性CYP 450和结合酶，将分子分泌到胆小管中，并维持与评价药物处置相关的肝脏特异性基因的高水平表达。该小型企业创新研究（SBIR）第二阶段项目的主要目标是进一步开发和优化这些微型人类肝脏培养物，并将其与小型化策略和分析技术相结合，以实现具有成本效益的高通量体外筛选。由于药物性肝损伤（DILI）是急性肝衰竭和药物高损耗率的主要原因，我们将优化我们的小型化人类肝脏，专门用于体外筛选基因型特异性和临床相关的药物处置和偶联DILI。我们在这里开发的技术可能会在开发几类治疗化合物（药物，生物制剂），评估环境毒物的处置和损伤潜力，肝脏生理学和疾病的基础研究，识别新的生物标志物以及肝病的个性化医学中找到广泛的用途。在未来，微技术与组织工程的持续结合可能会刺激其他组织模型的发展，并将其整合到所谓的“芯片上的人类”中。 公共卫生相关性：该项目中提出的研究旨在开发用于高通量筛选（HTS）应用的小型化人类肝脏微阵列，特别是用于评估药物处置和药物诱导的肝损伤，这对患者，监管机构和制药/生物技术行业来说是一个严峻的挑战。在未来，我们的微型微型肝脏HTS系统可能会在药物开发管道中更早地消除有问题的化合物，以减少患者接触不安全药物。我们在这里开发的技术也可能在评估环境毒物的损伤潜力，基础研究和肝病患者的个性化医疗中找到实用性。

DESCRIPTION (provided by applicant): Liver toxicity is the leading cause of pre-launch and post-market attrition of pharmaceutical compounds (i.e. Rezulin, Prexige). Significant species-specific differences in organ functions now necessitate supplementation of animal data with assays designed to assess human responses to drugs. Adherent cultures of primary human hepatocytes are considered to be the gold standard for evaluating preclinical drug metabolism, enzyme induction, and liver-specific toxicity. However, hepatocytes display a rapid (hours) decline in liver-specific functions under conventional culture conditions utilized routinely for drug development. Recently, a robust model of human liver tissue has been developed with optimized microscale architecture in an industry-standard multiwell format that retains liver-specific functions for 4-6 weeks in vitro. Hepatocytes in this microscale platform secrete liver-specific products, display functional CYP450 and conjugation enzymes, secrete molecules into the bile canaliculi, and maintain high levels of expression of liver-specific genes relevant for evaluating drug disposition. The primary objective of this Small Business Innovation Research (SBIR) Phase II project is to further develop and optimize these microscale human liver cultures and couple them with miniaturization strategies and assay technologies for cost-effective high-throughput in vitro screening. Since drug-induced liver injury (DILI) is a leading cause of acute liver failures and the high attrition rate of pharmaceuticals, we will optimize our miniaturized human livers specifically for the in vitro screening of genotype-specific and clinically-relevant drug disposition and coupled DILI. The technologies we develop here may find broad utility in the development of several classes of therapeutic compounds (drugs, biologics), in evaluating the disposition and injury potential of environmental toxicants, in fundamental investigations of liver physiology and disease, in the identification of new biomarkers, and in personalized medicine for liver disease. In the future, continued combination of microtechnology with tissue engineering may spur the development of other tissue models and their integration into the so-called 'human-on-a-chip'. PUBLIC HEALTH RELEVANCE: The studies proposed in this project are aimed towards developing a miniaturized human liver microarray for high-throughput screening (HTS) applications, specifically for evaluating drug disposition and drug-induced liver injury, a serious challenge for patients, regulatory agencies and the pharmaceutical/biotech industry. In the future, our miniaturized micro-liver HTS system may eliminate problematic compounds much earlier in the drug development pipeline towards reducing patient exposure to unsafe drugs. The technologies we develop here may also find utility in assessing the injury potential of environmental toxicants, in basic research, and in personalized medicine for patients with liver disease.