Role of CD164 in Skeletal Myogenesis

CD164 在骨骼肌生成中的作用

基本信息

批准号：
7193462
负责人：
Robert S. Krauss
金额：
$ 31.82万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-02-18 至 2008-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7193462
关键词：
Adhesions Affinity BHLH Protein Biochemical Biological Blood C-terminal Carbohydrates Cell Differentiation process Cell Line Cell Proliferation Regulation Cell Surface Proteins Cell surface Cells Chimeric Proteins Class Cysteine Cytoplasmic Tail Data Effector Cell Endosomes Enzymes Epitopes Family Feedback Genes Genetic Transcription Hematopoietic Leukocyte Trafficking Leukocytes Ligands Light Link Lysosomes Mediating Methods Molecular Genetics Mucin-2 Staining Method Mucins Mus Muscle Muscle Development Mutagenesis Mutation Myf-6 myogenic factor Myoblasts Myogenin Neuraminidase O-sialoglycoprotein endopeptidase Pattern Peptide Signal Sequences Phenotype Plasmids Play Polysaccharides Process Proliferating Proline Protein Overexpression Proteins Protocols documentation Rate Reagent Regulation Reporting Research Role Selectins Serine Skeletal Muscle Skeletal system Structure Testing Threonine Tissues Yeasts base cDNA Library carbohydrate structure expression cloning in vivo inhibitor/antagonist myocyte-specific enhancer-binding factor 2 myogenesis receptor research study sialomucin sialomucins transcription factor

项目摘要

DESCRIPTION (provided by applicant): Determination and differentiation of the skeletal muscle lineage is controlled by the MyoD family of myogenic transcription factors. These factors, along with the MEF-2 family of transcription factors, auto-activate and cross-activate the expression of each other, resulting in an autoregulatory, positive feedback network that maintains the myogenic phenotype. Determination and differentiation of cells in this lineage also require cell-cell contact between muscle precursors. However, the identities of the cell surface proteins involved in this phenomenon, and their specific roles in myogenesis, are not well understood. Sialomucins are cell surface proteins characterized by mucin domains, which are proline-, serine-, and threonine-rich regions on which a high percentage of the serine and threonine residues are O-glycosylated. These O-linked glycans are central to sialomucin function, as specific sialomucins are known to serve as high affinity ligands for selectins; selectins are transmembrane receptors that recognize sialylated carbohydrate structures on these sialomucins during leukocyte trafficking. The biological roles of sialomucins outside of leukocyte trafficking are poorly understood. We have identified Cd164 as a gene expressed in proliferating C2C12 myoblasts that is specifically upregulated during differentiation. Cd164 encodes a widely expressed cell surface sialomucin that has been implicated in regulation of cell proliferation, differentiation and adhesion of hematopoietic precursors. Stable overexpression of Cd164 enhances myoblast differentiation. Treatment of C2C12 cells with sialidase or O-sialoglycoprotease, two enzymes which destroy functional epitopes on CD164, also inhibits differentiation. Taken together, we hypothesize that CD164 is a likely effector of the need of cell-cell contact in myogenesis and that carbohydrate-based cell recognition may be involved in this process. The Specific Aims are: 1) to identify structural determinants of the pro-myogenic activity of CD164; 2) to identify counter-receptors for CD164; and 3) to investigate the role of CD164 in myogenesis in vivo. These experiments will provide information critical to molecular and genetic understanding of CD164's role in myogenesis. This research should therefore shed light on fundamental processes by which skeletal muscles develop.

描述（由申请人提供）：骨骼肌谱系的确定和分化由肌原性转录因子的Myod家族控制。这些因素以及MEF-2转录因子家族自动激活和交叉激活相互表达，从而导致自动调节的积极反馈网络维持肌源性表型。该谱系中细胞的测定和分化还需要肌肉前体之间的细胞细胞接触。然而，尚不清楚该现象中涉及的细胞表面蛋白的身份及其在肌发生中的特定作用。唾液蛋白是以粘蛋白结构域为特征的细胞表面蛋白，这些蛋白是脯氨酸，丝氨酸和苏氨酸富的区域，在该区域上，丝氨酸和苏氨酸残基很高的丝氨酸和苏氨酸残基是O-糖基化的。这些O连接的甘氨酸是唾液酸素功能的核心，因为已知特定的唾液蛋白被称为Selectins的高亲和力配体。 Selectins是跨膜受体，在白细胞运输过程中识别这些唾液酸蛋白上的硫化碳水化合物结构。人们对白细胞贩运之外的唾液核素的生物学作用知之甚少。我们已经将CD164鉴定为在分化过程中特异性上调的C2C12成肌细胞增殖的基因。 CD164编码了一种广泛表达的细胞表面唾液素蛋白，该细胞表面与细胞增殖，分化和造血前体的粘附有关。 CD164的稳定过表达增强了肌细胞的分化。用唾液酸酶或O-乳糖糖蛋白酶治疗C2C12细胞，两种酶破坏了CD164上的功能表位，也抑制了分化。综上所述，我们假设CD164可能是肌发生中细胞 - 细胞接触的可能效应因子，并且基于碳水化合物的细胞识别可能参与了这一过程。具体目的是：1）确定CD164促肌活性活性的结构决定因素； 2）识别CD164的反感应器； 3）研究CD164在体内肌发生中的作用。这些实验将提供对CD164在肌发生中作用的分子和遗传理解至关重要的信息。因此，这项研究应阐明骨骼肌发展的基本过程。