Hemodynamic Co-Culture Liver Model for Drug Discovery and Assessment

描述(申请人提供)：药物开发者迫切需要实验室中更好的工具来提高临床试验中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试验，但肝脏毒性仍然是药物失败的主要原因。HemoSher开发用于靶标识别的人类替代技术，并在发现后期和临床前早期阶段验证和筛选化合物的安全性和有效性。HemoSher已经开发出一种人体血管替代技术，用于识别和验证新的靶点，并筛选和选择进一步开发的最佳化合物(正在审查的美国专利60/879,710)。我们的人体血管替代设备独一无二地模拟了动脉粥样硬化早期阶段的血管解剖(包含内皮细胞和平滑肌细胞的共培养，由多孔膜隔开)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，并识别新的生物标记物和转录途径，以开发改进的药物治疗。基于细胞的替代模型正越来越多地被用于药物开发，以提供对人类反应的更准确的预测。这一第一阶段的SBIR应用程序建议使用HemoSher的专利平台技术开发肝脏血液动力学共培养模型，该模型将代表一个更优越的系统，用于筛选潜在的肝脏毒性药物，确定肝脏毒性的机制，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、单一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丧失，在药物开发过程中代表着无效的靶点识别和验证模型。与血管系统一样，重建对正常肝功能重要的解剖和生理特征对于有效的体外模型是必要的。例如，体内肝脏对内源性底物和外源性因素的生物反应依赖于不同类型的内皮细胞和上皮细胞之间的直接和间接‘串扰’。此外，它们的存活和基因表达谱在很大程度上依赖于局部血流动力学。因此，这项建议的总体目标是开发一种模拟体内生理学和血流动力学的大鼠肝脏共培养代理模型。该模型将由肝窦内皮细胞(SECs)和肝细胞共同培养组成，其中SECs暴露在正弦流体血流动力学中，在体内重建细胞表型，体外。如果成功，这一系统将使制药公司与目前的单一培养、静态系统相比，更好地了解药物疗效和临床前安全性/毒性的具体作用机制。此外，这项第一阶段SBIR的成功将导致第二阶段的应用，以开发更先进的人体肝毒性和炎症性疾病模型，纳入更多类型的细胞，例如枯否细胞和星状细胞。 与公共卫生相关：药物开发者迫切需要实验室中更好的工具来提高临床试验中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试验，但肝脏毒性仍然是药物失败的主要原因。HemoSher开发用于靶标识别的人类替代技术，并在发现后期和临床前早期阶段验证和筛选化合物的安全性和有效性。这一第一阶段的SBIR应用程序建议使用HemoSher的专利平台技术开发血流动力学大鼠肝窦内皮细胞和肝细胞共培养模型；这将是一个在筛选潜在的肝毒性药物、确定肝脏毒性机制和确定新的治疗靶点方面的优越系统。这项第一阶段SBIR的成功将导致第二阶段的应用，以开发更先进的人类肝毒性和炎症性疾病模型，包括更多类型的细胞，例如枯否细胞和星状细胞。

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.