Genetic dissection of the role of chondroitin sulfate in cartilage

硫酸软骨素在软骨中作用的基因解析

• 批准号: 8440194
• 负责人: YU YAMAGUCHI
• 金额: $ 43.88万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-15 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8440194
• 关键词: Abbreviations; Address; Affect; Alleles; Anabolism; Antioxidants; Biochemical; Biological; Biomechanics; Cartilage; Catabolism; Cell Death; Cell Survival; Cells; Charge; Chondrocytes; Chondroitin Sulfate Proteoglycan; Chondroitin Sulfates; Chronic; Collaborations; Core Protein; Data; Degenerative polyarthritis; Degradation Pathway; Dermatan Sulfate; Development; Diagnosis; Disease; Dissection; Event; Exhibits; Extracellular Matrix; Genes; Genetic; Genetic Models; Genetic Polymorphism; Health; Heparitin Sulfate; Human; In Vitro; Japan; Joints; Knock-out; Knockout Mice; Laboratories; Limb structure; Mechanical Stress; Mechanics; Metabolism; Minor; Modeling; Molecular; Mus; Osteoblasts; Pathogenesis; Pathology; Pathway interactions; Phase; Physiological; Play; Predisposing Factor; Prevention therapy; Property; Public Health; Reporting; Research; Right-On; Role; Signal Pathway; Signal Transduction; Skeletal system; Structure; Susceptibility Gene; Synovitis; Tendon structure; Tissues; Translating; Weight; age related; aggrecan; articular cartilage; base; bone; cell type; chondroitin synthase; editorial; genome wide association study; human disease; in vivo; insight; mouse model; neglect; notch protein; novel; polysulfated glycosaminoglycan; research study; response

DESCRIPTION (provided by applicant): Chondroitin sulfate (CS), a sulfated glycosaminoglycan, is one of the most abundant components of cartilage, comprising as much as 30% of the tissue. Because of its highly negative charge and sheer abundance, CS has long been assumed to be a key component to determine the unique biomechanical property of articular cartilage. Nevertheless, our understanding of the physiological role of CS in vivo and it involvement in human diseases is limited, mainly due to the lack of versatile mouse genetic models. To advance CS research to the next level, we have created a conditional null allele of Chsy1, the gene encoding chondroitin sulfate synthase 1. Remarkably, conditional Chsy1 knockout mice targeted to the limb skeletal system using Prx1-Cre exhibit joint pathologies strikingly similar to those of classical human osteoarthritis (OA). These observations, together with the previous in vitro observation that CS is protective to chondrocytes, led us to hypothesize that a decreased capability in CS synthesis underlie the pathogenesis of human OA. To examine this hypothesis, we propose the following studies: In Aim 1, we will determine the signaling basis of the degeneration of CS-deficient cartilage, focusing on the canonical Notch pathway. In Aim 2, we will employ an in vitro mechanical stress model to characterize cell death and antioxidant responses of CS-deficient chondrocytes to mechanical stress. In Aim 3, we will turn to additional conditional knockout experiments to further dissect chondrocyte-specific and osteoblast-specific roles of CS in the context of OA development. In Aim 4, collaborating with an expert in human association study, we will search for OA-associated, functional SNPs in human CHSY1 and other CS synthesizing genes. By these studies, we wish to gain novel insight into the molecular mechanisms of cartilage degeneration in OA, and to translate our observations in mice into the identification of novel susceptibility genes for human OA. PUBLIC HEALTH RELEVANCE: Osteoarthritis is a chronic debilitating disease that affects more than 20 million people in the US. This research will investigate the molecular mechanisms of cartilage damage seen in osteoarthritis using a novel genetic mouse model, and then translate the observation into human osteoarthritis by association study. Thus this project is considered directly as well as highly relevant to public health.

描述（由申请人提供）：硫酸软骨素（CS）是一种磺化糖胺聚糖，是软骨中最丰富的成分之一，占组织的30%。由于其高负电荷和丰富的数量，CS一直被认为是决定关节软骨独特生物力学特性的关键成分。然而，我们对CS在体内的生理作用及其在人类疾病中的作用的了解是有限的，主要是由于缺乏通用的小鼠遗传模型。为了将CS的研究推进到一个新的水平，我们创建了一个Chsy1的条件空等位基因，该基因编码硫酸软骨素合成酶1。值得注意的是，使用Prx1-Cre靶向肢体骨骼系统的条件Chsy1基因敲除小鼠表现出与经典人类骨关节炎（OA）惊人相似的关节病理。这些观察结果，加上之前的体外观察，CS对软骨细胞有保护作用，使我们假设CS合成能力的下降是人类OA发病的基础。为了验证这一假设，我们提出了以下研究：在Aim 1中，我们将确定cs缺陷软骨退行性变的信号基础，重点关注典型的Notch通路。在Aim 2中，我们将采用体外机械应力模型来表征cs缺陷软骨细胞对机械应力的细胞死亡和抗氧化反应。在Aim 3中，我们将转向额外的条件敲除实验，以进一步剖析CS在OA发展中的软骨细胞特异性和成骨细胞特异性作用。在Aim 4中，我们将与人类关联研究专家合作，在人类CHSY1和其他CS合成基因中寻找与oa相关的功能性snp。通过这些研究，我们希望对OA软骨退变的分子机制有新的认识，并将我们在小鼠中的观察结果转化为人类OA新的易感基因的鉴定。