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: 8742187
• 负责人: JOHN H. POSTLETHWAIT
• 金额: $ 67.58万
• 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/8742187
• 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; public health relevance; 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(由申请者提供)：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，通常是由于管道中的后期失败。尽管在VITR中，针对蛋白质或细胞系的筛选提供了重要的途径，但对完整的脊椎动物进行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公正的方法，可以筛选所有蛋白质和组织，其中许多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，利用全球基因表达模式来确定人类疾病的新疗法。幼鱼为完整的动物药物筛选提供了特别有用的对象，因为它们的器官功能类似于人类器官，包括用于吸收的上皮细胞、用于新陈代谢的肝脏、用于排泄的肾脏，以及对药物毒性敏感的眼睛、耳朵、大脑和甲状腺等器官。鱼可以直接从水中吸收药物，它们的小尺寸使得测试变得经济。疾病状态的特征是指示疾病病因和对疾病的反应的基因活动的星座。在一个创新的屏幕上，这个项目 将识别转录疾病信号(TDSS)，即其活性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使TDS恢复到健康鱼类的化合物。该项目预测，TDS的变化将比形态变化更早地揭示药物效应，并且将比体外基于蛋白质或细胞的测试更好地代表整个有机体对治疗的反应。结果包括改进了慢性病治疗的筛查和改进了药物筛选的动物模型。研究将使用两种截然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)的转基因青竹模型，以及范科尼贫血(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是使用 RNA-SEQ用于识别定义患病鱼与健康鱼的TDSS。目的2评价HTG-Edge和NanoStringnCounter在TDSS检测中的有效性。目标3和4是对青海MM(Aim 3)和FA斑马鱼(Aim 4)进行试点筛查，以确定将TDS从疾病状态转变为健康状态的化合物。HITS将被用于鱼类的机械分析，以及患有MM和FA疾病的哺乳动物的安全性和有效性研究。通过开发新的但广泛适用的药物筛选资源和方法，以及改进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，实现这些目标将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签名的创新概念来确定人类疾病的潜在疗法的最佳实践方案来加强研究基础设施，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加强翻译研究。