Joint degeneration: Somatic mosaic analysis in a transgenic mouse

关节退化：转基因小鼠的体细胞嵌合分析

• 批准号: 7136599
• 负责人: Stephanos Kyrkanides
• 金额: $ 19.5万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7136599
• 关键词: arthritis; genetically modified animals; joints; laboratory mouse; model; osteoarthritis; pathology; prostaglandin endoperoxide synthase; prostaglandins; transfection

DESCRIPTION (provided by applicant): Osteoarthritis (OA) is the most common form of arthritis and a major cause of activity limitation and physical disability in the elderly: 80%-90% of humans over the age of 65 suffer from OA. Available treatments for OA are mostly palliative with limited specificity. In fact, the recently identified serious side effects of some intra- oral COX-2 selective inhibitors (i.e. Vioxx, Bextra) underscore the need for the development of novel anti- inflammatory therapies for OA. The purpose of this research proposal is to characterize a mouse model of degenerative joint disease following the conditional induction of intra-articular, low level IL-1beta expression in the adult mouse and to explore the utility of siRNA-based anti-inflammatory therapy in the management of temporomandibular joint (TMJ) osteoarthritis. To this end, we will employ somatic mosaic analysis in the Col1-IL1beta-XAT transgenic mouse model recently developed in our laboratory, which utilizes a Cre/loxP based molecular genetic method to temporally and spatially activate a germline-transmitted recombinational substrate containing a dormant IL-1beta transcription unit. Preliminary data demonstrate the development of OA-like pathology in the knees and TMJ of Col1-IL1beta-XAT transgenic mice after intra-articular Cre administration as assessed by histopathological, immunohistochemical and behavioral studies. The following specific aims are proposed to further develop and characterize this model: (1) Characterize the development of TMJ arthritis following somatic mosaic analysis in the Col1- IL1beta-XAT transgenic mouse in terms of joint pathology and orofacial pain; (2) evaluate the effectiveness of intra-articular anti-mPGES-1 and anti-COX-2 siRNA therapy versus available pharmacologic inhibitors in the TMJ of Col1-IL1beta-XAT transgenic mice. We have recently developed small inhibitory RNA (siRNA) constructs capable of attenuating COX-2, as well as other members of the cyclooxygenase-prostaglandin pathway, including COX-1, mPGES-1 and cPGES. We propose to express these siRNA constructs intra-articulalry using a feline immuno-deficiency viral platform, FIV(siRNA). The proposed studies will contribute in the evaluation of novel therapeutic targets (i.e. mPGES- 1) as well as contribute to the development of new therapeutic tools (siRNA). Our proposal is submitted in response to the NIDCR program announcement PA-04-139 entitled "Joint degeneration: Mouse models", which solicits research proposals employing genetically defined and genetically modified mouse models to explore the biological mechanisms underlying OA and novel therapeutics such as siRNA technology. To this end, we have brought together a multidisciplinary team of experts from the University of Rochester that have successfully collaborated and published in the past and who will contribute the necessary expertise for the completion of this project.

描述（由申请人提供）：骨关节炎（OA）是关节炎的最常见形式，也是老年人活动受限和身体残疾的主要原因：80%-90%的65岁以上的人患有OA。现有的OA治疗大多是姑息性的，特异性有限。事实上，最近发现的一些口服考克斯-2选择性抑制剂（即万络，倍他乐）的严重副作用强调了开发新的OA抗炎疗法的必要性.本研究的目的是在成年小鼠关节内条件性诱导低水平IL-1 β表达后表征退行性关节疾病的小鼠模型，并探索基于siRNA的抗炎治疗在颞下颌关节（TMJ）骨关节炎管理中的实用性。为此，我们将在我们实验室最近开发的Col 1-IL-1 β-XAT转基因小鼠模型中采用体细胞嵌合体分析，该模型利用基于Cre/loxP的分子遗传学方法在时间和空间上激活含有休眠IL-1 β转录单位的种系传递重组底物。初步数据表明，通过组织病理学、免疫组织化学和行为学研究评估，关节内给予Cre后，Col 1-IL 1 β-XAT转基因小鼠膝关节和TMJ中OA样病理学的发展。本研究的主要目的是：（1）通过体细胞嵌合体分析，对Col 1-IL 1 β-XAT转基因小鼠TMJ关节炎的发生发展进行关节病理学和口面疼痛的表征;（2）评估关节内抗mPGES-1和抗COX-2 siRNA治疗与现有药物抑制剂相比在Col 1-IL 1 β-XAT转基因小鼠。我们最近开发了能够减弱考克斯-2以及环加氧酶-前列腺素途径的其他成员（包括考克斯-1、mPGES-1和cPGES）的小抑制性RNA（siRNA）构建体。我们建议使用猫免疫缺陷病毒平台FIV（siRNA）表达这些siRNA构建体。拟议的研究将有助于评价新的治疗靶点（即mPGES- 1），并有助于开发新的治疗工具（siRNA）。我们的提案是为了响应NIDCR项目公告PA-04-139“关节变性：小鼠模型”而提交的，该公告征求研究提案，采用遗传定义和遗传修饰的小鼠模型来探索OA的生物学机制和新疗法，如siRNA技术。为此，我们汇集了来自罗切斯特大学的多学科专家团队，他们过去曾成功合作并发表过论文，他们将为完成该项目贡献必要的专业知识。