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

基本信息

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
项目类别：
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

项目摘要

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) 是一种硫酸化糖胺聚糖，是软骨中最丰富的成分之一，占组织的 30%。由于其高度负电荷和丰富的丰度，CS长期以来一直被认为是决定关节软骨独特生物力学特性的关键组成部分。然而，我们对CS在体内的生理作用及其与人类疾病的关系的了解是有限的，这主要是由于缺乏通用的小鼠遗传模型。为了将 CS 研究推向新的水平，我们创建了 Chsy1 的条件无效等位基因，该基因编码硫酸软骨素合酶 1。值得注意的是，使用 Prx1-Cre 靶向肢体骨骼系统的条件 Chsy1 敲除小鼠表现出与那些小鼠惊人相似的关节病理学。典型的人类骨关节炎（OA）。这些观察结果，加上之前的体外观察结果，即 CS 对软骨细胞有保护作用，使我们推测 CS 合成能力的下降是人类 OA 发病机制的基础。为了检验这一假设，我们提出以下研究：在目标 1 中，我们将确定 CS 缺陷软骨退化的信号传导基础，重点关注典型的 Notch 通路。在目标 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。在目标 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 发展背景下的软骨细胞特异性和成骨细胞特异性作用。在目标 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，我们希望获得对 OA 软骨退化分子机制的新见解，并将我们在小鼠中的观察结果转化为鉴定人类 OA 的新易感基因。公共卫生相关性：骨关节炎是一种慢性衰弱性疾病，影响着超过 2000 万人的美国。这项研究将利用新型基因小鼠模型研究骨关节炎中软骨损伤的分子机制，然后通过关联研究将观察结果转化为人类骨关节炎。因此，该项目被认为与公共卫生直接且高度相关。