HIGH-THROUGHPUT IN VIVO SUBCELLULAR-RESOLUTION VERTEBRATE SCREENING PLATFORM

高通量体内亚细胞分辨率脊椎动物筛选平台

基本信息

批准号：
8477325
负责人：
Mehmet Fatih Yanik
金额：
$ 38.92万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-27 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8477325
关键词：
Animal Model Animals Biodistribution Biological Assay Biology Brain Cancer Biology Cardiovascular system Cell Line Cells Chemicals Communicable Diseases Complex Development Disease Disease Progression Disease model Embryo Endocrine system Environment Extracellular Matrix Failure Fiber Gene Silencing Genes Genetic Genetic Screening Goals Habitats Heart Image Immobilization Immune system In Vitro Injection of therapeutic agent Injury Kidney Larva Lead Liver Mammals Microfluidics Microinjections Microsurgery Modeling Natural regeneration Neoplasm Metastasis Nerve Degeneration Nervous System Physiology Neuraxis Neurobiology Neurons Optics Organ Pancreas Pathogenesis Pharmaceutical Preparations Pharmacologic Substance Phenotype Physiological Preclinical Drug Evaluation Process Resolution Specificity Speed Spinal cord injury Staging System Techniques Technology Testing Time Toxic effect Translational Research United States National Institutes of Health Vertebrates Vision Whole Organism Zebrafish cancer type cellular targeting chemical genetics cost drug candidate drug development fundamental research gene function genome-wide high throughput screening human disease in vivo innovation mutant regenerative screening small molecule

项目摘要

DESCRIPTION (provided by applicant): The ability to study whole organisms makes it possible to study complex in vivo processes that cannot be replicated in vitro such as organ development, liver, pancreas, heart, and neuronal regeneration, cancer metastasis, neural degeneration, infectious disease progression, pathogenesis, cardiovascular, immune, endocrine, and nervous system functions. Cells are not transformed and are in their normal physiological environment of cell-cell, extracellular matrix, and other interactions. Microarray studies and in vitro screens using cell lines and millions of combinatorially synthesized compounds have generated thousands of possible genetic targets and drug candidates. Identification of specificity, potency, toxicity, and biodistribution of pharmaceuticals as well as functions of thousands genes on entire organs like kidney, liver, heart, and brain cannot be done in vitro, and require use of in vivo animal models. Currently, there is significant gap between the throughput and capabilities of in vitro and in vivo assays on vertebrates. As a result, during early stages of drug screening and development, pharmaceuticals cannot be tested in vivo. Failure of tests on animals at later stages of development not only costs dearly, but also slows progress significantly. Yet, high-throughput testing of gene functions and compounds using in vivo vertebrate animal models has so far been significantly limited due to the absence of key technologies. Here, we propose a highly transformative technology that will allow, for the first time, large- scale in vivo genetic and chemical screens at cellular resolution on complex organs of vertebrates such as heart, liver, kidney, pancreas, vision, immune system, and central nervous system. This technology can impact a broad spectrum of fields ranging from neurobiology to regenerative biology, and cancer biology. The proposed high-speed whole-animal manipulation, orientation, immobilization, imaging, microsurgery, and injection platform will enable a dramatic increase in the throughput and complexity with which in vivo assays can be performed (~5-10 seconds per animal depending on the observed phenotype and manipulation complexity instead of the 10-30 minutes it currently takes). Our proposal is highly relevant to NIH's roadmap goals as it will allow systematic and unbiased genome-wide vertebrate studies to dramatically accelerate both fundamental and translational research in identification of gene functions as well as in discovery of drug leads. To demonstrate system capabilities, we will perform the first large-scale in vivo chemical screen for regenerating micro-surgically injured spinal-cord fibers.

描述（由申请人提供）：研究整个生物体的能力使得在体外过程中研究复杂的过程是可以在体外复制的复杂过程，例如器官发育，肝脏，胰腺，心脏和神经元再生，癌症转移，神经退化，感染性疾病进展，病原体，病原体，免疫，免疫，内部和神经系统的功能。细胞不会转化，并且处于细胞细胞，细胞外基质和其他相互作用的正常生理环境中。微阵列研究和体外筛选使用细胞系和数百万合成化合物的化合物产生了数千种可能的遗传靶标和候选药物。鉴定药物的特异性，效力，毒性和生物分布以及数千个基因在肾脏，肝脏，心脏和大脑等整个器官上的功能，不能在体外完成，并且需要使用体内动物模型。当前，体外和体内脊椎动物的吞吐量和能力之间存在显着差距。结果，在药物筛查和开发的早期阶段，不能在体内测试药物。在后来的发展阶段，对动物的测试失败不仅成本高昂，而且还会大大减慢进展。然而，由于缺乏关键技术，使用体内脊椎动物模型对基因功能和化合物进行了高通量测试已受到显着限制。在这里，我们提出了一种高度变革的技术，该技术将首次允许在脊椎动物的复杂器官（例如心脏，肝脏，肾脏，胰腺，胰腺，视觉，免疫系统和中枢神经系统）的复杂器官上进行大规模的体内遗传和化学筛选。这项技术可以影响从神经生物学到再生生物学和癌症生物学的广泛领域。提出的高速全动物操作，取向，固定，成像，显微手术和注射平台将使可以进行体内测定的吞吐量和复杂性急剧增加（根据观察到的表型和操纵的复杂性，而不是目前的10-30分钟，则可以进行体内测定法（〜5-10秒）。我们的建议与NIH的路线图目标高度相关，因为它将允许系统的和无偏的全基因组脊椎动物研究在鉴定基因功能以及发现药物铅的鉴定方面显着加速基本和转化研究。为了展示系统功能，我们将执行第一个大规模的体内化学筛选，用于再生微表面受伤的脊柱纤维。