A 3D osteoarthritis model targeting patient populations with high risk genetic polymorphisms

针对具有高风险遗传多态性的患者群体的 3D 骨关节炎模型

• 批准号: 9376249
• 负责人: Vincent P Willard
• 金额: $ 22.5万
• 依托单位: CYTEX THERAPEUTICS INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9376249
• 关键词: 3p21; Address; Adult; Affect; Age; Alpha Cell; Arthritis; Biochemical; Biological Assay; Cartilage; Cartilage Matrix; Cell Line; Cells; Chromosomes; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Degenerative polyarthritis; Development; Dimensions; Disease; Disease Progression; Economic Burden; Engineering; Environmental Risk Factor; Etiology; Exhibits; GDF5 gene; Generations; Genetic; Genetic Polymorphism; Genetic Predisposition to Disease; Genetic Screening; Genetic Variation; Genome; Goals; Human; In Vitro; Incidence; Inflammation Mediators; Inflammatory; Joints; Libraries; Link; Longevity; Measures; Mechanics; Mediating; Modeling; Obesity; Operative Surgical Procedures; Pain; Patients; Pharmaceutical Preparations; Phenotype; Population; Preclinical Drug Evaluation; Predisposition; Production; Progressive Disease; Property; Prosthesis; Protocols documentation; Replacement Arthroplasty; Risk; Risk Factors; Role; Single Nucleotide Polymorphism; Somatic Cell; Source; Stimulus; Synovial joint; System; Technology; Testing; Therapeutic; Tissue Engineering; Tissues; Treatment Protocols; United States; Variant; aging population; articular cartilage; base; cartilage degradation; cell type; curative treatments; cytokine; disability; drug testing; effective therapy; genetic profiling; genetic risk factor; genetic signature; genetic variant; genome editing; genome wide association study; high risk; in vitro Model; induced pluripotent stem cell; joint injury; mechanical properties; novel; palliative; patient population; personalized therapeutic; population based; reduce symptoms; repaired; response; screening; success; therapeutic effectiveness; treatment response; treatment strategy

Abstract Osteoarthritis (OA) is a degenerative joint disease that affects an estimated 30 million adults in the United States and results in an economic burden of over $130 billion per year. Although the burden of OA is immense, current non-surgical treatments are only palliative, and no disease-modifying OA drugs (DMOADs) presently exist to address the problem. This lack of success in identifying DMOADs is frequently attributed to the variable causes of OA initiation and the dearth of human cartilage available for screening potential DMOADs. To increase the likelihood of identifying DMOADs, we propose to study a segmented OA population based on defined genetic predisposition to OA development. To produce a nearly unlimited source of human cartilage for use in DMOAD screening, we will use induced pluripotent stem cells (iPSCs) as a cell source for cartilage tissue engineering. The goal of this project is to create a platform screening technology to identify the therapeutic requirements of OA-associated genetic risk factors. Our approach is to create a three-dimensional in vitro model of OA which utilizes iPSC lines that have been modified to contain defined genetic variations. In Aim 1 we will employ genome editing technology to generate OA-associated single-nucleotide polymorphisms (SNPs) in the genome of hiPSCs. Engineered cartilage formed from these edited cells will be characterized using biochemical and micromechanical assays and then treated with inflammatory cytokines to induce OA-like changes in the cartilage. The resulting in vitro OA model will be validated by measuring matrix degradation, loss of mechanical properties, and production of inflammatory mediators. In Aim 2, our iPSC-based model of OA will be transferred to a 96-well plate format to facilitate the development of an OA drug screening platform. A set of model therapeutics known to inhibit inflammatory degradation will be used to validate the sensitivity of the model and to define high-throughput readouts of OA progression. Finally, two libraries of novel bioactive compounds will be screened for their ability to slow OA-associated degradation in our in vitro model. This proposal will help elucidate the mechanism by which genetic variants result in increased risk for OA and will catalyze the development of tailored OA therapeutics by providing a platform technology for identifying therapeutic effectiveness based on defined risk factors.

摘要 骨关节炎（OA）是一种退行性关节疾病，在美国估计有3000万成年人受到影响。 每年造成超过1300亿美元的经济负担。虽然OA的负担是巨大的， 目前的非手术治疗仅是姑息性的，并且目前没有疾病修饰OA药物（DMOAD） 存在是为了解决这个问题。在确定DMOAD方面缺乏成功通常归因于变量 骨关节炎发病的原因和缺乏人类软骨可用于筛选潜在的DMOAD。到 为了增加识别DMOAD的可能性，我们建议研究基于以下内容的分段OA人群： 明确了OA发生的遗传易感性。为人类提供几乎无限的软骨来源， 在DMOAD筛选中，我们将使用诱导多能干细胞（iPSC）作为软骨的细胞来源。 组织工程学该项目的目标是创建一个平台筛选技术，以识别 OA相关遗传风险因素的治疗要求。我们的方法是创造一个三维的 OA的体外模型，其利用已被修饰以含有确定的遗传变异的iPSC系。在 目的1我们将利用基因组编辑技术产生OA相关的单核苷酸多态性 在hiPSC的基因组中存在SNP。由这些编辑的细胞形成的工程软骨将被表征 使用生物化学和微机械测定，然后用炎性细胞因子处理以诱导OA样 软骨的变化。将通过测量基质降解来验证所得体外OA模型， 机械性能的损失和炎症介质的产生。在目标2中，我们基于iPSC的 OA将被转移到96孔板格式，以促进OA药物筛选平台的开发。 将使用一组已知抑制炎症降解的模型治疗剂来验证以下的敏感性： 该模型和定义OA进展的高通量读数。最后，两个新的生物活性库 将在我们的体外模型中筛选化合物减缓OA相关降解的能力。这 这项提案将有助于阐明遗传变异导致OA风险增加的机制， 通过提供平台技术， 基于确定的风险因素的治疗效果。