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

• 批准号: 8636032
• 负责人: Sean Patrick Ryder
• 金额: $ 30.93万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8636032
• 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

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展需要专门的细胞与多能祖细胞区分开。初始细胞命运规范在胚胎发生早期。然而，许多成年组织维持祖细胞的细胞利基，可在损害时介导修复。越来越清楚的是，RNA结合蛋白在调节区分决定中起着重要作用。在早期胚胎尚未开始的早期胚胎中，在大脑中，高度分支的细胞形态需要调节距离细胞核的距离。在此提案中，我们描述了一种凝聚力的HTS分析开发策略，以鉴定调节细胞命运的RNA结合蛋白的小分子抑制剂。所描述的测定包括在体外测定中确定阻断RNA结合活性和基于细胞的测定的分子，以识别影响分化状态的化合物。一旦建立了HTS的准备和技术可行性，这些测定法将提交给NIH MLPCN，以进行高吞吐量筛查和探测识别。所鉴定的化合物将用于研究细胞和动物模型和提取物中细胞命运规范的分子机制。独立的结果将直接比较三种不同的RNA结合蛋白对小分子的敏感性。这种分析可能导致制定更有效的目标选择和图书馆设计策略。最后，这里要筛选的靶标与包括脑癌，多发性硬化症和寄生虫感染在内的人类疾病有关。因此，通过HTS鉴定的探针可能最终可以发展为疗法。

项目成果

Efficient generation of transgenic reporter strains and analysis of expression patterns in Caenorhabditis elegans using library MosSCI.

• DOI: 10.1002/dvdy.24426
• 发表时间: 2016-09
• 期刊: Developmental dynamics : an official publication of the American Association of Anatomists
• 作者: Kaymak E;Farley BM;Hay SA;Li C;Ho S;Hartman DJ;Ryder SP
• 通讯作者: Ryder SP