Hemodynamic Co-Culture Liver Model for Drug Discovery and Assessment

描述（由申请人提供）：药物开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。肝毒性仍然是药物失败的主要原因，尽管在替代动物物种中进行了广泛的临床前试验。在后期发现和早期临床前阶段，haemshear开发人类替代技术，用于目标识别和化合物的安全性和有效性的验证和筛选。hemshear开发了一种人体血管替代技术，用于鉴定和验证新靶点，以及筛选和选择进一步开发的最佳化合物（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了动脉粥样硬化早期的血管解剖（包含内皮细胞和平滑肌细胞的共培养，由多孔膜分开）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，以开发改进的药物治疗。基于细胞的替代模型越来越多地用于药物开发，以提供更准确的人类反应预测。这项I期SBIR申请建议使用hemshear的专有平台技术开发肝脏血流动力学共培养模型，该模型将代表一个非常优越的系统，用于筛选潜在的肝毒性药物，确定肝毒性机制并确定新的治疗靶点。众所周知，用于研究药物疗效和毒性的静态、单培养肝细胞模型不能预测体内反应，并且由于固有的分化表型损失，在药物开发过程中作为靶点识别和验证的无效模型。像血管系统一样，重建对正常肝功能重要的解剖和生理特征是建立有效的离体模型所必需的。例如，肝脏在体内对内源性底物和外源性因子的生物反应依赖于内皮和上皮不同细胞类型之间的直接和间接“串扰”。此外，它们的生存和基因表达谱在很大程度上依赖于局部血流动力学。因此，本研究的总体目标是开发一种模拟体内生理和血流动力学的大鼠肝脏共培养替代模型。该模型将由窦状内皮细胞（SECs）和肝细胞共同培养，其中SECs暴露于窦状流体血流动力学中，在体外重建体内细胞表型。如果成功，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，而不是目前单一的静态系统。此外，第一阶段SBIR的成功将导致第二阶段的申请，以开发更先进的肝毒性和炎症疾病的人类模型，纳入更多的细胞类型，如Kupffer和星状细胞。

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.