Hemodynamic Co-Culture Liver Model for Drug Discovery and Assessment

用于药物发现和评估的血流动力学共培养肝脏模型

• 批准号: 8059220
• 负责人: Brett R Blackman
• 金额: $ 19.08万
• 依托单位: HEMOSHEAR THERAPEUTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8059220
• 关键词: Acetaminophen; Address; Albumins; Anatomy; Animals; Atherosclerosis; Biological; Biological Assay; Biological Markers; Biological Models; Blood Vessels; Blood flow; Carbon Dioxide; Cell Differentiation process; Cell Line; Cells; Clinical Trials; Clofibrate; Coculture Techniques; Data; Detection; Development; Devices; Discontinuous Capillary; Disease; Drug toxicity; Electrons; Endothelial Cells; Endothelium; Environment; Enzymes; Epithelium; Evaluation; Exogenous Factors; Exposure to; Failure; Figs - dietary; Gene Expression; Genes; Goals; Hepatocyte; Hepatotoxicity; Human; Immunofluorescence Immunologic; Incubators; Inflammation; Inflammatory; Investigation; Kupffer Cells; Laboratories; Lead; Legal patent; Liquid substance; Liver; Membrane; Metabolic; Metabolism; Microscopic; Modeling; Molecular; Molecular Profiling; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacotherapy; Phase; Phenobarbital; Phenotype; Physiological; Physiology; Preclinical Drug Evaluation; Preclinical Testing; Procedures; Process; Rattus; Relative (related person); Reverse Transcriptase Polymerase Chain Reaction; Rodent; Safety; Sampling; Screening procedure; Side; Small Business Innovation Research Grant; Smooth Muscle Myocytes; Staging; System; Technology; Time; Tissues; Toxic effect; Validation; Xenobiotic Metabolism; base; cell type; chemokine; cytokine; cytotoxicity test; drug development; drug discovery; drug efficacy; hemodynamics; improved; in vivo; liver function; novel; polycarbonate; pre-clinical; preconditioning; response; stellate cell; success; tool

DESCRIPTION (provided by applicant): Drug developers desperately need better tools in the laboratory to improve the 90% failure rate of drugs in clinical trials. Liver toxicity is still the leading cause of drug failure, despite extensive preclinical testing in surrogate animal species. HemoShear develops human surrogate technologies for target ID and validation and screening of compounds for safety and efficacy in the late discovery and early pre-clinical stages. HemoShear has developed a human vascular surrogate technology for identification and validation of novel targets and for screening and selection of optimal compounds for further development (US Patent 60/879,710 under review). Our human vascular surrogate device uniquely mimics the vascular anatomy (co- culture containing endothelial and smooth muscle cells, separated by a porous membrane) and hemodynamic environment during the early stages of atherosclerosis. The device enables investigation of the cellular and molecular mechanisms of human atherosclerosis and the identification of novel biomarkers and transcriptional pathways for development of improved drug therapies. Cell-based surrogate models are being used increasingly during drug development to provide more accurate predictions of human responses. This Phase I SBIR application proposes to develop a hemodynamic co-culture model of the liver using HemoShear's proprietary platform technology that will represent a far superior system with which to screen drug potential for hepatotoxicity, determine mechanisms of liver toxicity and identify novel targets for therapy. It is widely known that static, monoculture hepatocyte models utilized to study drug efficacy and toxicity are not predictive of the in vivo response, and represent ineffective models for target identification and validation in the drug development process because of the inherent loss of differentiated phenotype. Like the vasculature, recreating anatomical and physiological features important to normal liver function is necessary for an effective ex vivo model. For example, the biological response of the liver in vivo to both endogenous substrates as well as exogenous factors is dependent on the direct and indirect 'cross-talk' between the different cell types of the endothelium and epithelium. In addition, their survival and gene expression profiles are largely dependent on the local hemodynamics. Thus, the overall goal of this proposal is to develop a rat liver co-culture surrogate model that mimics in vivo physiology and hemodynamics. The model will consist of sinusoidal endothelial cells (SECs) and hepatocytes in co-culture where the SECs are exposed to sinusoid fluid hemodynamics, recreating in vivo cell phenotypes, ex vivo. If successful, this system will allow pharmaceutical companies to better understand the specific mechanism-of-action for drug efficacy and pre-clinical safety/toxicity compared to current monoculture, static systems. Additionally, success of this Phase I SBIR will lead to a Phase II application to develop a more advanced human model of hepatotoxicity and inflammatory disease, incorporating additional cell types, e.g. Kupffer and stellate cells. PUBLIC HEALTH RELEVANCE: Drug developers desperately need better tools in the laboratory to improve the 90% failure rate of drugs in clinical trials. Liver toxicity is still the leading cause of drug failure, despite extensive preclinical testing in surrogate animal species. HemoShear develops human surrogate technologies for target ID and validation and screening of compounds for safety and efficacy in the late discovery and early pre-clinical stages. This Phase I SBIR application proposes to develop a hemodynamic rat sinusoidal endothelial cell and hepatocyte co-culture model of the liver using HemoShear's proprietary platform technology; which will represent a far superior system to screen drug potential for hepatotoxicity, determine mechanisms of liver toxicity and identify novel targets for therapy. Success of this Phase I SBIR will lead to a Phase II application to develop a more advanced human model of hepatotoxicity and inflammatory disease, incorporating additional cell types, e.g. Kupffer and stellate cells.

描述（申请人提供）：药物研发人员迫切需要实验室中有更好的工具来改善药物在临床试验中90%的失败率。尽管在替代动物物种中进行了广泛的临床前测试，但肝毒性仍然是药物失败的主要原因。 HemoShear 开发人类替代技术，用于目标识别以及化合物验证和筛选，以确保后期发现和早期临床前阶段的安全性和有效性。 HemoShear 开发了一种人体血管替代技术，用于识别和验证新靶点以及筛选和选择最佳化合物以进行进一步开发（美国专利 60/879,710 正在审查中）。我们的人体血管替代装置独特地模拟动脉粥样硬化早期阶段的血管解剖结构（包含内皮细胞和平滑肌细胞的共培养物，由多孔膜分隔）和血流动力学环境。该设备能够研究人类动脉粥样硬化的细胞和分子机制，并识别新的生物标志物和转录途径，以开发改进的药物疗法。基于细胞的替代模型在药物开发过程中得到越来越多的使用，以提供更准确的人类反应预测。这一 I 期 SBIR 申请计划使用 HemoShear 的专有平台技术开发肝脏的血流动力学共培养模型，该模型将代表一个极其优越的系统，可用于筛选药物的肝毒性潜力、确定肝毒性机制并确定新的治疗靶点。众所周知，用于研究药物功效和毒性的静态单一培养肝细胞模型不能预测体内反应，并且由于分化表型的固有损失，代表了药物开发过程中目标识别和验证的无效模型。与脉管系统一样，重建对正常肝功能重要的解剖和生理特征对于有效的离体模型是必要的。例如，体内肝脏对内源性底物和外源性因子的生物反应取决于内皮和上皮不同细胞类型之间的直接和间接“串扰”。此外，它们的存活和基因表达谱在很大程度上取决于局部血流动力学。因此，该提案的总体目标是开发模拟体内生理学和血流动力学的大鼠肝脏共培养替代模型。该模型将由共培养的正弦内皮细胞 (SEC) 和肝细胞组成，其中 SEC 暴露于正弦流体血流动力学，离体重现体内细胞表型。如果成功，与当前的单一培养静态系统相比，该系统将使制药公司能够更好地了解药物功效和临床前安全性/毒性的具体作用机制。此外，第一期 SBIR 的成功将导致第二期应用，以开发更先进的肝毒性和炎症性疾病人类模型，纳入其他细胞类型，例如枯否细胞和星状细胞。 公共健康相关性：药物研发人员迫切需要更好的实验室工具来改善临床试验中 90% 的药物失败率。尽管在替代动物物种中进行了广泛的临床前测试，但肝毒性仍然是药物失败的主要原因。 HemoShear 开发人类替代技术，用于目标识别以及化合物验证和筛选，以确保后期发现和早期临床前阶段的安全性和有效性。该 I 期 SBIR 申请计划使用 HemoShear 的专有平台技术开发血液动力学大鼠肝窦内皮细胞和肝细胞共培养肝脏模型；这将代表一个更加优越的系统，用于筛选药物的肝毒性潜力、确定肝毒性机制并确定新的治疗靶点。 I 期 SBIR 的成功将导致 II 期应用，以开发更先进的肝毒性和炎症性疾病人类模型，纳入其他细胞类型，例如枯否细胞和星状细胞。