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

药物性肝损伤（DILI）是患者、临床医生、FDA和制药行业关注的问题 作为临床药物损耗和上市后药物撤回的主要原因。根据FDA的说法，DILI 是过去50年来安全性相关药物停药的最常见原因。作为一个潜在 为了解决这个问题，IQ-MPS附属于国际创新和质量联盟， 药物开发部强调了对人类肝脏微生理系统（MPS）进行鉴定的必要性， DILI使用环境（CoU）以及一套实用的MPS工程和质量要求 执行标准作业程序。在正在进行的NCATS SBIR期间，Lena Biosciences（LB） 开发并商业化了符合这些标准的SLS标准化灌注器官组MPS 先决条件。该SBIR的最终目标是使MPS具有革命性的合成血红蛋白， 血液替代品，使用FDA CDER和IQ-MPS附属机构的指南用于DILI CoU。 我们最近的出版物（前。摩尔Biosci. 2020）表明，MPS恢复细胞的氧化代谢， 多样化的灌注3D肝脏模型，通过线粒体电子传递显著增加细胞呼吸 链，药物代谢所需的CYP 450氧化，以及整体代谢所需的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 功能细胞氧化还原稳态的OXPHOS-感受态模型将提供体内样细胞对以下的敏感性： 药物及其活性代谢物和自由基，以及药物诱导的氧化和亚硝化应激， 导致细胞抗氧化防御的丧失，用于DILI威胁的全面表征，定位 MPS充分满足DILI CoU的生物学鉴定先决条件。 虽然已经报道了导致DILI的许多因素，但迄今为止， 使用原代人细胞进行体外试验的这些因素，以及最常见的体外试验类型 与DILI预防有关。因此，在本SBIR中，我们将重点测试药物 工业界发现最难去风险，检查氧化细胞应激的作用，在细胞的序列 导致DILI的事件，使用一组 分析以分离触发物并识别因果关系，并解决时间和对数倍数变化， 生物标志物，包括FDA指定的用于临床探索的生物标志物，相对于溶媒对照和 与ALT和ALP（临床DILI生物标志物）同时升高的相关性，以分离出 低或中度ALT时的最高对数倍数变化和特异性。 为了成功地进行研究并确保项目的成功，我们组建了一个专家团队 在先进的，OXPHOS-感受态细胞培养（LB），和药物诱导的肝损伤的预测性筛选（博士。 Salman Khetani，UIC）。

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).