Development of Aquatic Model Resources for Therapeutic Screens

用于治疗筛选的水生模型资源的开发

• 批准号: 9121651
• 负责人: JOHN H. POSTLETHWAIT
• 金额: $ 18.41万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9121651
• 关键词: Adolescent; Age; Animal Disease Models; Animal Model; Animals; Biological Assay; Birth; Brain; Cell Line; Cell Proliferation; Cell Survival; Cells; Chemicals; Chronic Disease; Complement; Complex; Cultured Cells; Dermis; Development; Diabetes Mellitus; Disease; Drug toxicity; Ear; Embryo; Epidermis; Epithelium; Etiology; Excretory function; Eye; Failure; Fanconi' s Anemia; Fish Diseases; Fishes; Gene Expression Profile; Genes; Goals; Hair follicle structure; Health; Heart Diseases; Hematological Disease; Human; In Vitro; Individual; Investigation; Kidney; Larva; Lead; Liver; Longitudinal Studies; Malignant Neoplasms; Mammals; Methodology; Methods; Modeling; Morphology; Mus; Organ; Outcome; Pancytopenia; Patients; Pharmaceutical Preparations; Pharmacotherapy; Play; Preclinical Drug Evaluation; Proteins; Protocols documentation; Research Infrastructure; Research Personnel; Research Project Grants; Resources; Role; Safety; Schizophrenia; Skin Cancer; Stem cells; System; Technology; Testing; Therapeutic; Thyroid Gland; Time; Tissues; Toxic effect; Toxin; Transgenic Organisms; Translational Research; Vertebrates; Water; Whole Organism; Zebrafish; absorption; base; combat; cost; disease phenotype; drug efficacy; hatching; human disease; human tissue; improved; in vivo; innovation; liver metabolism; melanocyte; melanoma; mutant; nano-string; novel; response; screening; skin disorder; small molecule; teleost fish; tissue culture; transcriptome sequencing

DESCRIPTION (provided by applicant): Creating new drugs is an important national need, but a critical barrier is escalating cost, often due to late failures in the pipeline. Although in vitr screens targeting proteins or cell lines provide important avenues, in vivo screens on intact vertebrates can help overcome late-failure problems. Intact animal disease models provide an unbiased approach that screens all proteins and tissues, many of which may not yet be known to play a role in disease. The goal of this resource-related research project is to develop animal models and related materials for using global gene expression patterns to identify novel therapies for human disease. Larval fish provide especially useful subjects for intact-animal drug screens because they have organs that function like human organs, including epithelia for absorption, livers for metabolism, kidneys for excretion, and organs like eyes, ears, brain and thyroid sensitive to drug toxicity. Fish can absorb drugs directly from the water and their small size makes testing economical. Disease states are characterized by constellations of gene activity indicating disease etiology and response to disease. In an innovative screen, this project will identify transcriptional disease signatures (TDSs), suites of genes whose activities change in disease, and will develop efficient methods to identify compounds that return the TDS to that of healthy fish. The project predicts that TDS changes will reveal drug effects earlier than altered morphologies would and will represent a whole-organism response to therapy better than in vitro protein- or cell-based tests. Outcomes include improved screens for chronic disease therapeutics and improved animal models for drug screening. Investigations will use two disparate models for human disease: a transgenic medaka model for malignant melanoma (MM, a deadly human skin cancer), and a mutant zebrafish model for Fanconi anemia (FA, a disease of bone marrow failure that inhibits stem cell proliferation and survival). Aim 1 is to use RNA-seq to identify TDSs that define diseased vs. healthy fish. Aim 2 is to evaluate HTG-Edge and NanoString nCounter for their efficiency in assaying TDSs. Aims 3 and 4 are to conduct pilot screens on MM medaka (Aim 3) and FA zebrafish (Aim 4) to identify compounds that change the TDS from disease to healthy profiles. 'Hits' will be candidates for mechanistic analyses in fish and for safety and efficacy studies in mammals suffering MM and FA disease. Achieving these aims will have an enduring influence on translational research by developing novel but widely applicable drug screen resources and methodologies, as well as improving animal models of human disease and exploiting the advantages of intact-animal screens, thus complementing other screen systems. Results will enhance research infrastructure by outlining best practice protocols for using the innovative concept of transcriptome disease signatures to identify potential therapeutics for human disease and enhance translational research by providing lead compounds for chemical therapies for melanoma and Fanconi anemia.

描述（由申请人提供）：创造新药是一个重要的国家需求，但一个关键的障碍是不断上升的成本，往往是由于后期失败的管道。尽管靶向蛋白质或细胞系的体外筛选提供了重要的途径，但在完整脊椎动物上的体内筛选可以帮助克服后期失败的问题。完整的动物疾病模型提供了一种无偏见的方法，可以筛选所有蛋白质和组织，其中许多蛋白质和组织可能还不知道在疾病中发挥作用。这个与资源相关的研究项目的目标是开发动物模型和相关材料，用于使用全球基因表达模式来识别人类疾病的新疗法。幼鱼为完整动物药物筛选提供了特别有用的受试者，因为它们的器官功能与人体器官相似，包括用于吸收的上皮细胞，用于代谢的肝脏，用于排泄的肾脏，以及对药物毒性敏感的器官，如眼睛，耳朵，大脑和甲状腺。鱼可以直接从水中吸收药物，它们的体积小，使测试经济。疾病状态的特征在于指示疾病病因和对疾病的反应的基因活性的星座。在一个创新的屏幕上，这个项目 将确定转录疾病签名（TDS），其活动在疾病中发生变化的基因套件，并将开发有效的方法来识别将TDS恢复到健康鱼类的化合物。该项目预测，TDS变化将比改变形态更早地揭示药物作用，并且将比基于体外蛋白质或细胞的测试更好地代表整个生物体对治疗的反应。成果包括改善慢性疾病治疗的筛选和改善药物筛选的动物模型。研究将使用两种不同的人类疾病模型：恶性黑色素瘤（MM，一种致命的人类皮肤癌）的转基因青鳉模型，以及范可尼贫血症（FA，一种抑制干细胞增殖和存活的骨髓衰竭疾病）的突变斑马鱼模型。目标1：使用 RNA-seq以鉴定定义患病鱼与健康鱼的TDS。目的2是评估HTG-Edge和NanoString nCounter在测定TDS中的效率。目的3和4是对MM青鳉（目的3）和FA斑马鱼（目的4）进行试点筛选，以鉴定将TDS从疾病改变为健康概况的化合物。“命中”将是鱼类机制分析以及患有MM和FA疾病的哺乳动物安全性和有效性研究的候选者。实现这些目标将通过开发新的但广泛适用的药物筛选资源和方法，以及改善人类疾病的动物模型和利用完整动物筛选的优势，从而补充其他筛选系统，对转化研究产生持久的影响。结果将通过概述使用转录组疾病特征的创新概念的最佳实践方案来增强研究基础设施，以确定人类疾病的潜在治疗方法，并通过为黑色素瘤和范可尼贫血的化学疗法提供先导化合物来增强转化研究。