Comparative HTS to identify small molecule effectors of cell fate specification

比较 HTS 来识别细胞命运规范的小分子效应物

基本信息

批准号：
8449117
负责人：
Sean Patrick Ryder
金额：
$ 29.84万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8449117
关键词：
Adult Affinity Animal Model Animals Binding Binding Sites Biological Biological Assay Brain Brain Neoplasms Caenorhabditis elegans Cell Extracts Cell Nucleus Cell model Cells Cellular Morphology Chemicals Colon Colon Adenocarcinoma Colorectal Complex Cultured Cells Cytoplasmic Granules Demyelinating Diseases Development Developmental Process Disease Effector Cell Embryo Embryonic Development Epithelial Exploratory/Developmental Grant for Diagnostic Cancer Imaging Fluorescence Polarization Gene Expression Genetic Models Genetic Transcription Genetic Translation Germ Cells Glial Cell Proliferation Glioblastoma Heterogeneous-Nuclear Ribonucleoprotein K Homologous Gene Housing Human Institutes Intervention KH Domain Lead Libraries Location Malignant Neoplasms Malignant neoplasm of brain Measures Mediating Messenger RNA Molecular Monitor Multiple Sclerosis Muscle N-terminal Nematoda Nematode infections Neurons Oligodendroglia Organ Outcome Parasitic infection Parasitic nematode Pharyngeal structure Play Post-Transcriptional Regulation Predisposition Protein Family Proteins RNA RNA Binding RNA Processing RNA Recognition Motif RNA Stability RNA-Binding Proteins Readiness Regulation Reproduction Research Rheumatoid Arthritis Role Specificity Stem cells Stress Tertiary Protein Structure Therapeutic Tissues To specify Transcript Translations United States National Institutes of Health Work Writing Zinc Fingers assay development base cell fate specification comparative cytokine design follow-up genetic regulatory protein high throughput screening human disease in vitro Assay in vivo inhibitor/antagonist interest mRNA Stability medulloblastoma novel therapeutics oocyte maturation pluripotency progenitor repaired response small molecule success tool tumor

项目摘要

DESCRIPTION (provided by applicant): Development of complex tissues and organs requires specialized cells to differentiate from pluripotent progenitors. Initial cell fate specification ocurs early in embryogenesis. However, many adult tissues maintain a niche of progenitor cells that mediate repair upon damage. It is becoming increasingly clear that RNA-binding proteins play an important role in regulating the decision to differentiate. This is particularly true in early embryos, where nascent transcription has not yet begun, and in the brain, where the highly branched cellular morphology necessitates regulation at a distance from the nucleus. In this proposal, we describe a cohesive HTS assay development strategy to identify small molecule inhibitors of the RNA-binding proteins that regulate cell fate. The assays described include in vitro assays to identify molecules that block RNA-binding activity and cell-based assays to identify compounds that effect differentiation state. Once HTS-readiness and technical feasibility are established, these assays will be submitted to the NIH MLPCN for high throughput screening and probe identification. The compounds identified will be used to study the molecular mechanisms of cell fate specification in cell and animal models and in extracts. An independent outcome will be a direct comparison of the susceptibility of three different RNA- binding protein families to small molecules. This analysis may lead to development of more efficient target selection and library design strategy. Finally, the targets to be screened here are relevant to human diseases including brain cancer, multiple sclerosis, and parasitic infection. As such, it is possible that probes identified through HTS could eventually be developed into therapies.

描述（由申请人提供）：复杂组织和器官的发育需要专门的细胞从多能祖细胞中分化出来。初始细胞命运规范发生在胚胎发生的早期。然而，许多成体组织都维持着祖细胞的生态位，这些祖细胞可以介导损伤后的修复。越来越清楚的是，RNA 结合蛋白在调节分化决策中发挥着重要作用。这在早期胚胎中尤其如此，在早期胚胎中，新生转录尚未开始，在大脑中，高度分支的细胞形态需要在远离细胞核的地方进行调节。在本提案中，我们描述了一种内聚 HTS 测定开发策略，以鉴定调节细胞命运的 RNA 结合蛋白的小分子抑制剂。所描述的测定包括鉴定阻断RNA结合活性的分子的体外测定和鉴定影响分化状态的化合物的基于细胞的测定。一旦 HTS 准备就绪和技术可行性确定，这些检测将提交给 NIH MLPCN 进行高通量筛选和探针鉴定。鉴定出的化合物将用于研究细胞和动物模型以及提取物中细胞命运规范的分子机制。一个独立的结果将是直接比较三个不同 RNA 结合蛋白家族对小分子的敏感性。该分析可能会导致开发更有效的目标选择和文库设计策略。最后，这里要筛选的靶点与人类疾病相关，包括脑癌、多发性硬化症和寄生虫感染。因此，通过 HTS 鉴定的探针最终有可能开发成治疗方法。