CAREER: Towards a stem cell-derived 3D human liver array for high-throughput screening

职业：开发用于高通量筛选的干细胞衍生 3D 人类肝脏阵列

• 批准号: 1557348
• 负责人: Salman Khetani
• 金额: $ 36.79万
• 依托单位: University of Illinois at Chicago
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1557348&HistoricalAwards=false
• 关键词: CAREER; Towards; stem; cell; derived

1351909 Khetani, Salman R.Drug-induced liver injury (DILI) is a leading cause of acute liver failures and liver transplants in humans that can lead to restrictions on drug use, black-box warnings on drugs, and the pre-launch and post-market attrition of pharmaceuticals. In addition to its impact on the well-being of patients, the economic impact of DILI is significant (~$1B and 12-15 years to bring 1 successful drug to market). Animal testing that is required by the Food and Drug Administration (FDA) is not fully representative of the human condition due to significant and evolutionary differences between animals and humans in liver functions. Therefore, preclinical screening of compounds for potential toxicity using human cells now constitutes a critical part of the drug development pipeline. Induced pluripotent stem cell-derived human liver cells, called hepatocytes (iHeps), can not only provide a near unlimited source of cells for screening millions of compounds, but also enable personalized (patient-specific) drug screening and therapeutic applications in the future. However, iHeps created to date are not as functionally mature as adult primary hepatocytes (isolated from donor livers) which is required to adequately predict clinical outcomes before the drugs are tested on live patients in clinical trials. To address this technology gap, the investigator will utilize microfabrication tools adapted from the semiconductor (microchip) industry to engineer a miniaturized 3D human liver mimic using iHeps and other liver supportive cell types that displays functional maturity and stability for several weeks in vitro (outside the body). This liver mimic will be characterized using several important markers for its functional maturity relative to adult primary hepatocytes. Additionally, the liver mimic will be probed for its ability to provide clinically meaningful information on DILI due to single drug and multiple drug treatments in attempts to model clinical drug dosing regimens. The investigator will integrate his research with educational programs designed to build a continuum of mentorship and scientific community in the nascent biomedical engineering (BME) undergraduate program at Colorado State University (CSU) and in the Northern Colorado region. In particular, the investigator will design and implement: a) a new course that exposes CSU BME college students to mentored research experiences in a formal research laboratory setting; b) a step-wise program that exposes high school students to career options in BME and culminates in hands-on summer research experiences in CSU BME laboratories; and, c) recurrent learning modules that expose K-6 students in an after-school program to advances in science and engineering. The iHep-based 3D human liver mimic that will be developed could ultimately lead to the development of efficacious and safe therapeutics earlier in drug development towards increasing the likelihood of clinical success and limiting patient exposure to unsafe drugs. This platform may also be applicable for evaluating the injury potential of industrial chemicals that contaminate the environment, in promoting a better cellular/molecular understanding of diseases of the liver, and in helping to design and implement personalized medicine strategies. More broadly, the engineered device could ultimately be integrated with other tissue models into a single integrated system that provides a multi-organ (i.e. 'whole body') level of understanding of various exogenous insults. The aforementioned educational efforts will develop a robust mechanism to get students of different age groups interested in science and engineering education and to get each other involved in teaching and implementing programs for younger students.Due to the interdisciplinary nature of the project, this CAREER award by the Biotechnology, Biochemical, and Biomass Engineering Program of the CBET Division is co-funded by the Instrument Development for Biological Research Program of the Division of Biological Infrastructure.

1351909 Khetani，Salman R.药物性肝损伤（DILI）是人类急性肝功能衰竭和肝移植的主要原因，可能导致药物使用限制、药物黑盒警告以及药物上市前和上市后的损耗。 除了对患者健康的影响外，DILI的经济影响也很显著（约10亿美元和12-15年才能将1种成功的药物推向市场）。美国食品药品监督管理局（FDA）要求的动物试验并不能完全代表人类的状况，因为动物和人类在肝功能方面存在显著的进化差异。因此，使用人类细胞对化合物进行潜在毒性的临床前筛选现在构成了药物开发管道的关键部分。诱导多能干细胞衍生的人肝细胞，称为肝细胞（iHep），不仅可以提供几乎无限的细胞来源，用于筛选数百万种化合物，而且还可以在未来实现个性化（患者特异性）药物筛选和治疗应用。然而，迄今为止创建的iHep在功能上不如成人原代肝细胞（从供体肝脏分离）成熟，这需要在临床试验中对活体患者进行药物测试之前充分预测临床结果。为了解决这一技术差距，研究人员将利用半导体（微芯片）行业的微制造工具，使用iHep和其他肝脏支持细胞类型设计小型化3D人类肝脏模拟物，这些细胞在体外（体外）显示功能成熟和稳定性数周。将使用几种重要标志物表征该肝脏模拟物相对于成年原代肝细胞的功能成熟度。此外，将探索肝脏模拟物提供关于由于单一药物和多种药物治疗导致的DILI的临床有意义信息的能力，以尝试模拟临床药物给药方案。调查员将整合他的研究与教育计划，旨在建立一个连续的导师和科学界在新生的生物医学工程（BME）本科课程在科罗拉多州立大学（CSU）和北方科罗拉多地区。特别是，调查员将设计和实施：a）一个新的课程，使CSU BME大学生在正式的研究实验室环境中获得指导性的研究经验; B）一个逐步的计划，使高中生在BME中获得职业选择，并在CSU BME实验室的实践夏季研究经验中达到高潮;以及，c）经常性学习模块，使K-6学生在课后计划中了解科学和工程的进步。即将开发的基于iHep的3D人类肝脏模拟物可能最终导致在药物开发早期开发有效和安全的治疗方法，以增加临床成功的可能性并限制患者暴露于不安全的药物。该平台还可用于评估污染环境的工业化学品的潜在伤害，促进对肝脏疾病更好的细胞/分子理解，并帮助设计和实施个性化医疗策略。更广泛地说，工程装置最终可以与其他组织模型集成到单个集成系统中，该系统提供对各种外源性损伤的多器官（即“全身”）水平的理解。上述教育工作将建立一个强大的机制，让不同年龄段的学生对科学和工程教育感兴趣，并让彼此参与教学和实施针对年轻学生的计划。由于该项目的跨学科性质，生物技术，生物化学，CBET部门的生物质工程项目由生物基础设施部门的生物研究项目仪器开发共同资助。