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：
10214526
负责人：
YUANYUAN no ZHANG
金额：
$ 23.25万
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 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10214526
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 Disease Dose Drug Evaluation Drug toxicity Evaluation Exhibits Genetic Transcription Goals Growth HIV HepG2 Hepatocyte Human In Vitro Individual Injury Institutes Integrase Inhibitors 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

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 Disease Dose Drug Evaluation Drug toxicity Evaluation Exhibits Genetic Transcription Goals Growth HIV HepG2 Hepatocyte Human In Vitro Individual Injury Institutes Integrase Inhibitors 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

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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

概括抗逆转录病毒疗法（ART）显着降低了与HIV相关的发病率和死亡率。但是，治疗艾滋病毒患者的预期寿命增加，艺术的长期安全吸引了人们越来越多的关注。那是非常重要的抗逆转录病毒方案的长期安全性。 2D培养中的人肝细胞细胞系数最多用于评估ART诱导的短期线粒体毒性（MTT）。但是，这些不能用于检测艺术引起的慢性MTT，因为它们无法生存超过毒性测试期间的几天。此外，抗病毒毒性筛查通常需要昂贵的初级例如2D培养系统中的外周血单核细胞。超过90％的药物通过 2D培养临床前研究无法满足随后所需的所需效率或安全边缘试验。显然，2D文化导致MTT筛选测定的预测能力不佳尽管在动物模型中经常进行长期毒性测试，但在较早的临床试验表明，动物毒理学研究可能不适合预测抗病毒的MTT 旨在人类使用。显然，晚期MTT没有足够的2D细胞培养或动物模型用于临床前药物开发。因此，迫切需要在毒性上发展更多以及临床预测性的体外评估，以评估MTT后期潜力的化合物。我们是为了证明茎/祖细胞存在于人尿中，即尿液衍生的干细胞（USC）。这些可以使用简单，非侵入性和低成本程序获得。 USC Express端粒酶活性具有强大的增殖物潜力。我们最近透露，USC的3D文化提供了长期的微环体的细胞生长和增殖，模仿体内条件，这可能具有改善临床前抗病毒药物研究的预测结果。那，是为了开发人类USC的3D培养系统，以进行艺术引起的MTT测试。我们假设人类 USC在3D器官中保持端粒酶活性和线粒体功能，这大大延伸了细胞培养系统的生活，可以长期评估晚期艺术引起的MTT。为了检验这一假设，我们提出以下提议：目标1。评估人类USC的干性特性和线粒体功能与USC的2D培养相比，长期培养的3D器官；目标2。确定细胞毒性，在3D USC培养物中抑制POL-γ，线粒体DNA含量和9抗逆转录病毒药物的MTT谱。我们测试的抗逆转录病毒药物将对3D培养系统中的MTT产生影响，并进行排名基于体外MTT分析的有效浓度。因此，使用患者衍生从尿液到产生3D培养评估的细胞为抗逆转录病毒药物开发提供了有希望的工具在评估抗HIV药物的MTT中的个性化医学。艺术的益处通常受到延迟与药物相关的毒性的限制。作为有监视器通常肝细胞 14 细胞体外临床体外这毒品评估相关的第一的细胞和更好的t 潜在这个项目的目标预料根据干和