Astrocyte-specific ligand discovery by phage display

通过噬菌体展示发现星形胶质细胞特异性配体

• 批准号: 8995701
• 负责人: Philip J Horner
• 金额: $ 26.1万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8995701
• 关键词: Affinity; Animal Model; Astrobiology; Astrocytes; Award; Bacteriophages; Base Sequence; Behavior; Binding; Biological Markers; Biology; Biomedical Engineering; Brain; Cell Separation; Cell physiology; Cell surface; Cells; Cellular biology; Cognition; Complex; Confocal Microscopy; Data; Development; Disease; Enzymes; Evaluation; Exhibits; Flow Cytometry; Fluorescence; Genetic Markers; Glial Fibrillary Acidic Protein; Goals; Health; Heterogeneity; Homeostasis; Image; Immunity; In Vitro; Knowledge; Label; Lead; Libraries; Ligands; Locomotion; Mediating; Metabolic; Methodology; Molecular; Molecular Probes; Mus; Nerve Degeneration; Nervous System Trauma; Neuraxis; Neurobiology; Neuroglia; Neuronal Plasticity; Neurons; Neurotransmitters; Oligodendroglia; Outcome; Pathway interactions; Peptide Phage Display Library; Peptides; Phage Display; Play; Population; Preclinical Drug Evaluation; Process; Property; Publishing; Recording of previous events; Reporter; Research; Research Personnel; Role; Sensory; Slice; Sorting - Cell Movement; Specificity; Spinal Cord; Technology; Testing; Work; astrocyte progenitor; base; biomarker identification; design; drug discovery; experience; gliogenesis; in vitro testing; in vivo; in vivo imaging; macroglia; macrophage; molecular imaging; molecular marker; neurotransmission; novel; protein aminoacid sequence; receptor; screening; tool

 DESCRIPTION (provided by applicant): Astrocytes have been proposed to play critical roles in plasticity as well as neuronal degeneration. Despite their abundance and key homeostatic roles very little is known regarding how astrocytes are generated and the mechanisms whereby astrocytes achieve specialized functions. In contrast, there are over 130 sub-classes of neurons and the transcriptional network regarding many of these lineages have been discovered. This is in part due to the significant emphasis that has been placed on understanding neurons but also due to the number of molecular and genetic markers that have been identified for different classes of neurons. The latter has facilitated the development of lineage specific tools and animal models to decipher the various roles of neuronal subtypes in divergent behaviors such as cognition, locomotion and sensory function. Here we propose to utilize a phage display screening approach to identify unique molecular aspects of astrocytes and astrocyte subtypes in order to accelerate our understanding of astrocyte function in health and disease. Cell-based phage screening is a powerful technology that involves the selection of peptide ligands from over 2 billion unique candidate peptide sequences based on desired cell binding properties and without a priori knowledge of specific receptors. We will combine our expertise in gliogenesis and glial cell biology with that of bioengineering, drug screening and discovery to develop novel tools for subclassifying astrocytes and also for direct imaging of astrocytes in vivo. We propose: Aim 1. Identification of astrocyte-specific peptides by phage display library selection, Aim 2. Evaluation of astrocyte-specific peptides as cell-specific markers for astrocytic populations and Aim 3. Development of a molecular probe for in vivo imaging of astrocytes. This proposal takes advantage of the unique research capabilities of the two principle investigators and their long-standing collaborative history. We expect that the tools and technologies developed through this proposed project will enable rapid advances in glial biology through the identification of unique biomarkers of astrocytes in general as well as astrocyte subtypes classified by function.

 描述（由申请人提供）：已提出星形胶质细胞在可塑性和神经元变性中起关键作用。尽管它们的丰富和关键的稳态作用，但关于星形胶质细胞如何产生以及星形胶质细胞实现专门功能的机制知之甚少。相比之下，有超过130个神经元子类，并且已经发现了关于这些谱系中的许多谱系的转录网络。这部分是由于对理解神经元的重视，但也由于已为不同类别的神经元确定的分子和遗传标记的数量。后者促进了谱系特异性工具和动物模型的发展，以破译神经元亚型在不同行为（如认知、运动和感觉功能）中的各种作用。在这里，我们建议利用噬菌体展示筛选方法来识别星形胶质细胞和星形胶质细胞亚型的独特分子方面，以加速我们对星形胶质细胞在健康和疾病中功能的理解。基于细胞的噬菌体筛选是一种强大的技术，其涉及基于所需的细胞结合特性从超过20亿个独特的候选肽序列中选择肽配体，而无需特异性受体的先验知识。我们将联合收割机结合我们在胶质细胞生成和胶质细胞生物学方面的专业知识，以及生物工程、药物筛选和发现方面的专业知识，开发用于星形胶质细胞亚分类和星形胶质细胞体内直接成像的新工具。我们建议：通过噬菌体展示文库选择鉴定星形胶质细胞特异性肽，目的2。评估星形胶质细胞特异性肽作为星形胶质细胞群体的细胞特异性标志物和Aim 3。星形胶质细胞活体成像分子探针的研制。该提案利用了两位主要研究人员的独特研究能力及其长期合作历史。我们预计，通过这个拟议的项目开发的工具和技术将通过识别一般星形胶质细胞的独特生物标志物以及按功能分类的星形胶质细胞亚型，使胶质细胞生物学的快速发展成为可能。