Drug Discovery for Multiple Hereditary Exostoses

多种遗传性外生骨疣的药物发现

基本信息

批准号：
8735612
负责人：
Jeffrey D Esko
金额：
$ 37.7万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-16 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8735612
关键词：
Abbreviations Affect Biological Assay Cartilage Cartilaginous exostosis Cataloging Catalogs Cell surface Cells Cellular biology Chemicals Chinese Hamster Chinese Hamster Ovary Cell Chondrocytes Collaborations Data Analyses Defect Development Dimethyl Sulfoxide Disaccharides Disease Disease model Dose Drug Formulations Drug Kinetics Elements Enzymes Epiphysial cartilage Exostoses Family Femur Fibroblast Growth Factor Flow Cytometry Frequencies Generations Genes Genomics Growth Factor Heparan Sulfate Biosynthesis Heparitin Sulfate Hereditary Multiple Exostoses Human Image Image Analysis Informatics Institutes Lead Libraries Link Liquid Chromatography Mass Spectrum Analysis Measures Mesenchymal Metabolism Molecular Weight Morphologic artifacts Mus Mutation Natural History Outcome Ovary Patients Pharmaceutical Chemistry Pharmaceutical Preparations Phenocopy Phycoerythrin Physiology Pilot Projects Rare Diseases Reducing Agents Signal Pathway Signal Transduction Source Structure Testing Therapeutic Toxic effect Toxicology Validation analog base cheminformatics drug candidate drug discovery high throughput screening human disease in vivo innovation insight long bone monolayer neglect pre-clinical public health relevance research study response restoration rib bone structure screening skeletal stem

项目摘要

DESCRIPTION (provided by applicant): Multiple Hereditary Exostoses (MHE) is an autosomal dominant disorder characterized by the formation of ectopic cartilage-capped growth plate-like exostoses next to long bones and other skeletal elements. MHE results from mutations in the genes Ext1 or Ext2, which diminish the capacity of cells in the growth plate and the surrounding perichondrium to make heparan sulfate. The mechanism by which a change in heparan sulfate content causes ectopic osteochondromas is unknown, but evidence suggests that the decrease in heparan sulfate affects multiple signaling pathways through which growth factors regulate the organization and proliferation of chondrocytes in the growth plate. Regardless of the mechanism, the primary defect is in the assembly of heparan sulfate, suggesting that restoring the level of heparan sulfate would diminish the frequency of exostoses. All cells make heparan sulfate through a common mechanism. Thus, we propose to use Chinese hamster ovary (CHO) cells that are functionally hemizygous for Ext1 and to employ a primary cell- based screen to find potential drug candidates that augment heparan sulfate expression. Pilot studies have been done in collaboration with the High Content Screening Core in the Conrad Prebys Center for Chemical Genomics at the Sanford-Burnham Institute. Assay optimization, validation and final implementation of the proposed image-based high-throughput screening assay will be accomplished. The Cheminformatics and Informatics Core at Sanford-Burnham will assist in data analysis, artifact filtering, replicate hit confirmation, and generation of dose response profiles. Secondary assays will test positive hits for their impact on heparan sulfate content and structure. Tertiary assays will measure if the hits modulate heparan sulfate expression in mouse chondrocytes and perichondrial cells and in human chondrocytes. The resultant rank ordering of potency of confirmed hit sets and chemotypes merged with additional secondary and tertiary assay results will aid in hit-to-lead identification. The Cheminformatics Core will search for commercially available analogs to support limited structure-activity profiling. Agents that enhance heparan sulfate synthesis will be evaluated by formulation, stability, pharmacokinetics, and toxicity and their capacity to reduce exostoses in Ext1+/-;Ext2+/- mice. The central hypothesis is that altering key enzymes involved in heparan sulfate metabolism can result in restoration of functionally normal levels of heparan sulfate and reduction of exostoses in mice, which would serve as a proof-of-principle for pharmacological manipulation of exostosis formation in MHE patients.

描述（由申请人提供）：多个遗传外遗体（MHE）是一种常染色体显性疾病，其特征是形成了异位软骨限制的生长板状的外遗体，旁边是长骨和其他骨骼元素。 MHE是由Ext1或Ext2基因突变引起的，这会降低生长板中细胞和周围的perichondrium中硫酸乙酰肝素的能力。硫酸乙酰肝素含量的变化引起异位骨软骨瘤的机制尚不清楚，但证据表明，乙酰硫酸乙酰肝素硫酸盐的下降会影响多种信号通路，从而调节生长板中软骨细胞的组织和增殖。无论机制如何，主要缺陷都是在硫酸乙酰肝素的组装中，这表明恢复硫酸乙酰肝素的水平会降低外骨的频率。所有细胞都通过一种共同的机制使硫酸乙酰肝素使硫酸盐。因此，我们建议使用用于Ext1功能上半合子的中国仓鼠卵巢（CHO）细胞，并采用基于原始细胞的筛选来找到增强硫酸乙酰肝素表达的潜在药物候选物。试点研究是与Sanford-Burnham Institute的Conrad Prebys化学基因组学中心的高素质筛选核心合作进行的。将完成测定优化，验证和最终实施，对基于图像的高通量筛选测定法进行。 Sanford-Burnham的化学信息学和信息学核心将有助于数据分析，人工制品过滤，重复命中确认以及剂量反应概况的产生。次要测定将测试其对硫酸乙酰肝素含量和结构的影响。如果命中率调节小鼠软骨细胞和围缘细胞和人软骨细胞中的heparan硫酸盐表达，则第三次分析将测量。确认的命中集的效力和额外的二级和三级测定结果合并的确认命中集和化学型的效力将有助于命中率识别。化学信息学核心将搜索市售的类似物，以支持有限的结构活动分析。增强硫酸乙酰肝素合成的药物将通过配方，稳定性，药代动力学和毒性及其减少Ext1 +/-; ext2 +/-鼠标减少外骨的能力进行评估。中心假设是，改变参与硫酸乙酰肝素代谢的关键酶会导致硫酸乙酰肝素的功能正常水平恢复小鼠的雌雄同体，这将是MHE患者中外胞症形成的药理学操纵的原则证明。