3D Culture Systems Of Urine-Derived Stem Cell For NTRI-Induced Mitotoxicity Assessment

用于 NTRI 诱导的细胞毒性评估的尿源干细胞 3D 培养系统

基本信息

批准号：
10083026
负责人：
YUANYUAN no ZHANG
金额：
$ 19.38万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-10 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10083026
关键词：
3-Dimensional Accounting Affect Animal Model Animals Anti-HIV Agents Anti-Retroviral Agents Antiviral Agents Attention Biological Assay Biological Sciences Cell Culture System Cell Culture Techniques Cell Differentiation process Cell Line Cell Proliferation Cell Survival Cells Chronic Clinic Clinical Clinical Trials Collaborations Detection Development Disease Dose Drug Evaluation Drug Screening Drug toxicity Environment Evaluation Exhibits Genetic Transcription Goals Growth HIV HepG2 Hepatocyte Human In Vitro Individual Injury Institutes Integrase Life Life Expectancy Longevity Measures Metabolism Mitochondria Mitochondrial DNA Modeling Monitor Morbidity - disease rate Organoids Patients Pharmaceutical Preparations Pharmacology Phenotype Physiological Ploidies Preclinical Drug Development Preclinical Drug Evaluation Primary Cell Cultures Property Regimen Respiration Safety System Telomerase Tenofovir Testing Therapeutic Tissue Banks Toxic effect Toxicity Tests Toxicology Urine Zidovudine antiretroviral therapy base cell growth clinical predictors clinically relevant cost cost effective cytotoxicity drug development drug testing early phase clinical trial human stem cells human tissue improved in vitro Assay in vivo insight monocyte mortality outcome prediction peripheral blood personalized medicine pre-clinical preclinical study preclinical toxicity procedure cost progenitor screening side effect stem stem cells stemness three dimensional cell culture tool two-dimensional

项目摘要

Summary Antiretroviral therapy (ART) has significantly reduced HIV-related morbidity and mortality. However, therapeutic life expectancy for individuals with HIV increased, the long-term safety of ART has gained increasing attention. Thus, it is highly important to the long-term safety of antiretroviral regimens. Human hepatocyte cell lines in 2D culture are most used to evaluate short-term mitochondrial toxicity (MtT) induced by ART. However, these cannot be used in detection of ART-induced chronic MtT because they cannot survive more than days during toxicity testing. In addition, antiviral drug toxicity screening often requires expensive primary such as peripheral blood monocytes in 2D culture systems. More than 90% of drugs that pass through in 2D culture preclinical studies fail to meet the desired efficacy or safety margins required in subsequent trials. Clearly, 2D cultures have contributed to the poor predictive power of MtT screening assays in . Although long-term toxicity tests are often performed in animal models, the high rates of MtT observed in earlier clinical trials suggest that animal toxicology studies may not be suitable for predicting MtT of antiviral intended for human use. Apparently, there are no adequate 2D cell culture or animal models for late MtT for preclinical drug development. Hence, there is an urgent need to develop more toxicologically and clinically predictive in vitro assays to assess compounds for the potential of late MtT. We were the to demonstrate that stem/progenitor cells exist in human urine, i.e., urine-derived stem cells (USC). These can be obtained using simple, non-invasive and low-cost procedures. USC express telomerase activity possess robust proliferative potential. We recently revealed that 3D culture of USC provides a long-term, microenvironmen for cell growth and proliferation, mimicking in vivo conditions, which may have to improve the predictive outcome of preclinical antiviral drug studies. Thus, is to develop 3D culture systems of human USC for ART-induced MtT testing. We hypothesize that human USC maintain telomerase activity and mitochondrial function in 3D organoids, which considerably extends the life of cell culture systems, enabling long-term assessment of late ART-induced MtT . To test this hypothesis, we propose the following: Aim 1. Assess the stemness properties and mitochondrial function of human USC in 3D organoids over long-term culture, compared to 2D culture of USC; Aim 2. Determine the cytotoxicity, inhibition of Pol-γ, mitochondrial DNA content and MtT profiles of 9 antiretroviral drugs in 3D USC cultures. We that the antiretroviral drugs tested will exhibit effects on MtT in the 3D culture systems and be ranked to their effective concentrations based on in vitro MtT assay. Therefore, the use of patients-derived cells from urine to generate 3D culture assay offers a promising tool for antiretroviral drug development personalized medicine in the assessment of MtT of anti-HIV drugs. benefit of ART is often limited by delayed drug-associated toxicity. As has monitor commonly hepatocytes 14 cells vitro clinical vitro the drugs evaluation relevant first cells and better t potential the goal of this project anticipate according stem and

概括抗逆转录病毒治疗（ART）显着降低了艾滋病毒相关的发病率和死亡率。艾滋病毒感染者的预期寿命因此，ART 的长期安全性越来越受到关注。二维培养的人肝细胞系的长期安全性最高。用于评估 ART 引起的短期线粒体毒性 (MtT)。不能用于检测 ART 诱导的慢性 MtT，因为它们的存活时间不能超过此外，抗病毒药物毒性筛查通常需要昂贵的初步费用。例如2D培养系统中的外周血单核细胞，90%以上的药物都在其中通过。 2D 培养临床前研究未能满足后续所需的疗效或安全裕度显然，2D 培养导致 MtT 筛选试验的预测能力较差。尽管长期毒性试验经常在动物模型中进行，但在动物模型中观察到的高 MtT 发生率早期的临床试验表明动物毒理学研究可能不适合预测抗病毒药物的 MtT 显然，没有足够的 2D 细胞培养物或动物模型用于晚期 MtT。因此，迫切需要进行更多的毒理学研究。以及临床预测性体外测定，以评估化合物对晚期 MtT 的潜力。证明人尿液中存在干/祖细胞，即尿源干细胞（USC）。可以使用简单、非侵入性且低成本的程序获得 USC 表达端粒酶活性。我们最近透露，南加州大学的 3D 文化提供了长期的、细胞生长和增殖的微环境，模仿体内条件，这可能有提高临床前抗病毒药物研究的预测结果。旨在开发人类 USC 3D 培养系统，用于 ART 诱导的 MtT 测试。 USC 在 3D 类器官中保持端粒酶活性和线粒体功能，这大大扩展了细胞培养系统的寿命，从而能够对晚期 ART 诱导的 MtT 进行长期评估。我们提出以下建议：目标 1. 评估人类 USC 的干性特性和线粒体功能长期培养的 3D 类器官与 USC 的 2D 培养相比。目标 2. 确定细胞毒性， 3D USC 培养物中 Pol-γ 的抑制、线粒体 DNA 含量和 9 种抗逆转录病毒药物的 MtT 谱。将测试的抗逆转录病毒药物在 3D 培养系统中表现出对 MtT 的影响并进行排名其有效浓度基于体外 MtT 测定，因此使用患者衍生的。从尿液中提取细胞生成 3D 培养测定为抗逆转录病毒药物开发提供了一种有前景的工具个性化医疗在抗 HIV 药物 MtT 评估中的应用 ART 的益处常常受到延迟药物相关毒性的限制。有监视器通常肝细胞 14 细胞体外临床体外这药物评估相关的第一的细胞和更好该项目的潜在目标预料根据干和