Development of Aquatic Model Resources for Therapeutic Screens

用于治疗筛选的水生模型资源的开发

基本信息

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
项目类别：
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&apos 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

项目摘要

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）：创造新药是国家的一项重要需求，但一个关键障碍是成本不断上升，这通常是由于管道后期失败造成的。尽管针对蛋白质或细胞系的体外筛选提供了重要途径，但对完整脊椎动物的体内筛选可以帮助克服后期失败问题。完整的动物疾病模型提供了一种筛选所有蛋白质和组织的公正方法，其中许多蛋白质和组织可能尚不清楚在疾病中发挥作用。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，以利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，因为它们具有像人体器官一样功能的器官，包括用于吸收的上皮、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，而且它们的体型较小，使得测试变得经济。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星座。在一个创新的屏幕上，这个项目将识别转录疾病特征（TDS），即在疾病中活性发生变化的基因组，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 的变化将比形态改变更早地揭示药物效应，并且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。成果包括改进慢性病治疗方法的筛选和改进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：用于恶性黑色素瘤（MM，一种致命的人类皮肤癌）的转基因青鳉模型，以及用于范可尼贫血（FA，一种抑制干细胞增殖和骨髓衰竭的疾病）的突变斑马鱼模型。生存）。目标1是使用 RNA-seq 用于识别 TDS，从而定义患病鱼与健康鱼。目标 2 是评估 HTG-Edge 和 NanoString nCounter 在测定 TDS 方面的效率。目标 3 和 4 是对 MM 青鳉鱼（目标 3）和 FA 斑马鱼（目标 4）进行试点筛选，以确定将 TDS 从疾病状态转变为健康状态的化合物。 “Hits”将成为鱼类机械分析以及患有 MM 和 FA 疾病的哺乳动物安全性和有效性研究的候选药物。通过开发新颖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影响。结果将通过概述使用转录组疾病特征的创新概念来识别人类疾病的潜在疗法的最佳实践方案来增强研究基础设施，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增强转化研究。