A microphysiological system with a synthetic hemoglobin, Blood Substitute, for mechanistic assessment of drug-induced liver injury

具有合成血红蛋白（血液替代品）的微生理系统，用于药物性肝损伤的机械评估

• 批准号: 10385048
• 负责人: Jelena Vukasinovic
• 金额: $ 22.11万
• 依托单位: LENA BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10385048
• 关键词: 3-Dimensional; Accounting; Acetaminophen; Active Biological Transport; Acute Liver Failure; Affect; Animals; Antioxidants; Bile Acids; Bile Acids and Salts; Biological; Biological Assay; Biological Markers; Biological Sciences; Biotechnology; Blood; Blood Substitutes; Cell Culture Techniques; Cell Death; Cell Membrane Proteins; Cell Respiration; Cell membrane; Cell model; Cell physiology; Cells; Cellular Stress; Cholestasis; Clinical; Complex; Consensus; Development; Drug Industry; Drug Screening; Drug usage; Electron Transport; Energy-Generating Resources; Engineering; Ensure; Enzymes; Erythrocytes; Etiology; Event; Foundations; Free Radicals; Galactose; Glucose; Glutathione; Goals; Heme Group; Hemochromatosis; Hemoglobin; HepG2; Hepatocyte; Hepatotoxicity; Homeostasis; Hospitalization; Human; Immune; In Vitro; Incidence; Industry; Injury; International; Iron; Lead; Lipid Peroxidation; Liver; Marketing; Mediating; Membrane; Membrane Lipids; Methodology; Mitochondria; Modeling; N-acetyl-4-benzoquinoneimine; National Center for Advancing Translational Sciences; Organ; Organelles; Oxidation-Reduction; Oxidative Phosphorylation; Oxidative Stress; Oxygen; Parents; Pathway interactions; Patients; Perfusion; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Positioning Attribute; Prevention; Procedures; Production; Publications; Pump; Reactive Oxygen Species; Reperfusion Injury; Reporting; Respiration; Risk; Role; Running; Safety; Small Business Innovation Research Grant; Specificity; Standardization; Stress; T-Cell Activation; T-Lymphocyte; Testing; Tissues; Toxic effect; Triage; adaptive immunity; adverse drug reaction; analog; base; bile salts; cell growth; cell injury; clinical decision-making; cytokine; cytotoxicity; drug induced liver injury; drug metabolism; drug testing; drug withdrawal; extracellular vesicles; glutathione peroxidase; heme a; human model; in vitro Assay; in vitro testing; in vivo; inhibitor; injury prevention; innovation; insight; liver injury; liver ischemia; liver transplantation; microphysiology system; nitrosative stress; oxidation; peripheral blood; predictive test; response; success

Drug induced liver injury (DILI) is a concern for patients, clinicians, the FDA, and the pharmaceutical industry as the leading cause of clinical drug attrition and post-marketing drug withdrawals. According to the FDA, DILI has been the most frequent cause of safety-related drug withdrawals for the past 50 years. As a potential solution to the problem, the IQ-MPS Affiliate to the International Consortium for Innovation and Quality in Pharmaceutical Development highlighted the need to qualify human liver microphysiological systems (MPS) for DILI context of use (CoU) with a set of pragmatic engineering and quality requirements for the MPS implementation into standard operating procedures. During an ongoing NCATS SBIR, Lena Biosciences (LB) developed and commercialized an SLAS-standardized, Perfused Organ Panel MPS that meets these prerequisites. The ultimate goal of this SBIR is to qualify the MPS with a revolutionary synthetic hemoglobin, Blood Substitute, for DILI CoU using the guidance of the FDA CDER and the IQ-MPS Affiliate. Our recent publication (Front. Mol. Biosci. 2020) shows that the MPS restores cellular oxidative metabolism in diverse, perfused, 3D liver models, significantly increasing cell respiration by mitochondrial electron transport chain, CYP450 oxidation required for metabolism of drugs, and OXPHOS ATP production required for holistic cell function, and all cell processes from active transport of molecules across the cell membrane to organelle function. An OXPHOS-competent model of cellular redox homeostasis will provide in vivo-like cell sensitivity to drugs and their reactive metabolites and free radicals, and drug-induced oxidative and nitrosative stress that leads to the loss of cellular antioxidant defense for comprehensive characterization of DILI threats, positioning the MPS to adequately meet biological qualification prerequisites for DILI CoU. While numerous factors contributing to DILI have been reported, to date there is no consensus on the rank of these factors for in vitro testing using primary human cells, and on the types of in vitro assays that are the most relevant for DILI prevention. Therefore, in this SBIR we will focus on testing those compounds that the drug industry found the most difficult to de-risk, examine the role of oxidative cell stress in the sequence of cellular events that lead to DILI, provide mechanism-informative insight into the DILI sequelae using a battery of assays to isolate the trigger(s) and identify causalities, and resolve temporal and log-fold change in biomarkers, including the FDA-designated biomarkers for clinical exploration, relative to vehicle controls and in relation with the coinciding rise of ALT and ALP, clinical DILI biomarkers, in order to isolate those with the highest log-fold change and specificity at low or moderate ALT. To successfully carry out the studies and ensure the project’s success, we have assembled a team of experts in advanced, OXPHOS-competent cell cultures (LB), and predictive screening of drug-induced livery injury (Dr. Salman Khetani, UIC).

药物性肝损伤 (DILI) 是患者、新来者、FDA 和制药行业关注的问题 据 FDA 称，DILI 是临床药物消耗和上市后药物撤回的主要原因。 过去 50 年来，它一直是安全相关药物停药的最常见原因。 为了解决这一问题，IQ-MPS 隶属于国际创新与质量联盟 药物开发强调需要验证人类肝脏微生理系统（MPS）的资格 DILI 使用环境 (CoU) 以及 MPS 的一组实用工程和质量要求 在正在进行的 NCATS SBIR 中，Lena Biosciences (LB) 实施了标准操作程序。 开发并商业化了 SLAS 标准化的灌注器官面板 MPS，以满足这些要求 该 SBIR 的最终目标是通过革命性的合成血红蛋白来鉴定 MPS， 血液替代品，用于 DILI CoU，使用 FDA CDER 和 IQ-MPS 附属机构的指导。 我们最近发表的文章（Front. Mol. Biosci. 2020）表明 MPS 可恢复细胞氧化代谢 多样化、灌注的 3D 肝脏模型，通过线粒体电子传输显着增加细胞呼吸 链、药物代谢所需的 CYP450 氧化以及整体所需的 OXPHOS ATP 产生 细胞功能，以及从分子主动转运穿过细胞膜到细胞器的所有细胞过程 具有 OXPHOS 功能的细胞氧化还原稳态模型将提供类似体内的细胞敏感性。 药物及其反应性代谢物和自由基，以及药物诱导的氧化和亚硝化应激 导致细胞抗氧化防御能力丧失，无法全面表征 DILI 威胁、定位 MPS 充分满足 DILI CoU 的生物学资格先决条件。 尽管已经报道了许多导致 DILI 的因素，但迄今为止，对于 DILI 的排名尚未达成共识。 使用原代人类细胞进行体外测试的这些因素，以及最重要的体外测定类型 与 DILI 预防相关，因此，在本 SBIR 中，我们将重点测试该药物的那些化合物。 业界发现最难消除风险的是，检查氧化细胞应激在细胞序列中的作用 导致 DILI 的事件，使用一系列方法提供对 DILI 后遗症的机制信息洞察 分离触发因素并确定因果关系并解决时间和对数倍数变化的分析 生物标志物，包括 FDA 指定的用于临床探索的生物标志物，相对于载体对照和 与 ALT 和 ALP（临床 DILI 生物标志物）同时升高的关系，以便分离出那些 低或中等 ALT 时具有最高的对数倍数变化和特异性。 为了顺利开展研究并确保项目成功，我们组建了专家团队 先进的 OXPHOS 感受态细胞培养物 (LB) 以及药物引起的肝损伤的预测性筛查（Dr. 萨尔曼·赫塔尼 (Salman Khetani，UIC)。