Cbfβ mediates articular cartilage regeneration and repair in aging

Cbfβ 介导衰老过程中的关节软骨再生和修复

• 批准号: 9564595
• 负责人: Wei Chen
• 金额: $ 37.13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9564595
• 关键词: Adult; Affect; Age; Aging; American; Animal Model; Arthritis; Binding Sites; Biological Assay; Bone Diseases; Candidate Disease Gene; Cartilage; Cell Lineage; Cells; Chondrocytes; Chondrogenesis; Complex; Core-Binding Factor; Cytomegalovirus; Data; Degenerative polyarthritis; Development; Disease; Female; Gene Expression; Gene Expression Profile; Genes; Genetic Predisposition to Disease; Goals; Hip region structure; Homeostasis; Immunologist; In Vitro; Injection of therapeutic agent; Interdisciplinary Study; Investigation; Joints; Knee; Knee Osteoarthritis; Knock-out; Knockout Mice; Light; Luciferases; Mediating; Mesenchymal Stem Cells; Metabolic Bone Diseases; Mus; Natural regeneration; Nature; Operative Surgical Procedures; Osteoblasts; Pain; Pain management; Pathogenesis; Pathologic; Patients; Phenotype; Physiological; Plant Roots; Play; RNA Interference; Regulation; Research; Role; Shoulder; Signal Pathway; Signal Transduction; Skeletal Development; Subfamily lentivirinae; Synovial Membrane; System; Tamoxifen; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; Vertebral column; WNT Signaling Pathway; aged; aging population; articular cartilage; base; beta catenin; bone; bone cell; cartilage degradation; cartilage regeneration; cartilage repair; chromatin immunoprecipitation; design; early onset; gain of function; improved; insight; knock-down; male; mechanical load; mouse model; novel strategies; overexpression; postnatal; prevent; promoter; skeletal disorder; small hairpin RNA; subchondral bone; transcription factor

The long term goal of this study is to understand the mechanisms mediating osteoarthritis in aging. The specific goal of this study is to characterize the mechanism underlying how Cbfβ mediates cartilage regeneration and repair in osteoarthritis. Osteoarthritis (OA) is the most common form of arthritis affecting the knees, hips and spines, inflicting pain and physical limitation on over 70% of Americans between the age of 55 and 70. Current therapeutic options for OA are still limited to pain management and surgical intervention representing a significant concern in the aging population. Recent studies have shed light on the nature of OA genetic susceptibility and confirmed a number of candidate genes involved in the damage of the synovium, articular cartilage, and subchondral bone in OA pathogenesis, including Wnt signaling. However, the root causes of the disease remain unclear. Having investigated the gene expression patterns between OA patients and normal donors, and in combining with the known regulation data, Qingyou et al. identified a number of transcriptional factors and other genes which may play important roles in the development of OA including Core-binding factor beta (Cbfβ). The PI’s lab has extensively study Cbfβ function in osteoblasts and chondrocytes during mouse skeletal development. Nevertheless, the function of Cbfβ in OA pathogenesis and articular cartilage regeneration and repair remains unclear. In order to study OA pathogenesis in chondrocyte- specific Cbfβ deficient mice, we generated the inducible conditional knockout (CKO) Cbfβf/fCol2α1-CreER mice using Tamoxifen injections. We discovered that Cbfβ CKO mice developed spontaneous OA at the age of 3.5 months, showing severe OA phenotype at the shoulders, knees, hips and spines. Our data demonstrated that Wnt canonical signaling was down-regulated and Yap expression was up-regulated in Cbfβf/fCol2α1-CreER mice and that Cbfβ overexpression mediated by AAV-CMV-Cbfβ has significant protection against OA. Based on our preliminary data, we hypothesize that deficiency of Cbfβ is one of the main causes of cartilage degeneration in OA and aging and that overexpression of Cbfβ enhances cartilage regeneration and repair in OA by regulating Wnt signaling and Yap signaling. We will test this hypothesis through three specific aims. In Aim 1, we will determine the roles of Cbfβ in aging articular chondrocyte homeostasis through extensive phenotypic analyses of adult and aged, female and male Cbfβf/fCol2α1-CreER mice in physiological and pathological conditions. In Aim 2, we will define the function of Cbfβ in cartilage regeneration and repair and preventing OA genesis in aging by characterizing Cbfβ gain-of-function mouse model. We will dissect the mechanism underlying how Cbfβ regulates Wnt and Yap signaling during chondrogenesis and articular cartilage regeneration and repair in Aim 3. The proposed study will provide important insights into the roles of Cbfβ in OA by elucidating the mechanism by which Cbfβ regulates Wnt and Yap signaling in articular cartilage regeneration and repair. A multidisciplinary research team been established to achieve the research goals.

本研究的长期目标是了解衰老过程中骨关节炎的介导机制。的 本研究的具体目标是表征Cbfβ如何介导软骨的潜在机制 骨关节炎的再生和修复。骨关节炎（OA）是影响关节炎的最常见形式， 膝盖，臀部和脊柱，造成疼痛和身体限制超过70%的美国人之间的55岁 和70.目前OA的治疗选择仍然局限于疼痛管理和手术干预 这在老龄化人口中是一个值得关注的问题。最近的研究揭示了OA的本质 遗传易感性，并确认了一些参与滑膜损伤的候选基因， 关节软骨和软骨下骨在OA发病机制中的作用，包括Wnt信号传导。然而，根 这种疾病的病因尚不清楚。研究了OA患者和正常人之间的基因表达模式， 和正常供体，结合已知的调控数据，Qingyou等鉴定了一些 转录因子和其他基因，可能发挥重要作用的发展，OA，包括 核心结合因子β（Cbfβ）。PI的实验室广泛研究了Cbfβ在成骨细胞中的功能， 小鼠骨骼发育过程中的软骨细胞。然而，Cbfβ在OA发病机制中的作用和 关节软骨的再生和修复仍不清楚。为了研究OA在软骨细胞中的发病机制， 特异性Cbfβ缺陷小鼠，我们产生了诱导型条件性敲除（CKO）Cbfβf/fCol 2 α1-CreER小鼠 使用他莫昔芬注射。我们发现Cbfβ CKO小鼠在3.5岁时发生自发性OA 月，在肩、膝、髋和脊柱处显示严重OA表型。我们的数据表明， 在Cbfβf/fCol 2 α1-CreER中，Wnt经典信号转导下调，雅普表达上调 AAV-CMV-Cbfβ介导的Cbfβ过表达对OA具有明显的保护作用。 根据我们的初步数据，我们假设Cbfβ缺乏是软骨损伤的主要原因之一， Cbfβ的过度表达增强了骨关节炎软骨的再生和修复， 通过调节Wnt信号和雅普信号来调节OA。我们将通过三个具体目标来检验这一假设。在 目的1、通过广泛的体外实验，研究Cbfβ在老化关节软骨细胞内环境稳定中的作用。 成年和老年、雌性和雄性Cbfβf/fCol 2 α1-CreER小鼠在生理和 病理条件。在目标2中，我们将定义Cbfβ在软骨再生和修复中的功能， 通过表征Cbfβ功能获得性小鼠模型预防衰老中的OA发生。我们将解剖 Cbfβ在软骨形成和关节软骨形成过程中调节Wnt和雅普信号的机制 Aim 3中的软骨再生和修复。拟议的研究将为以下方面的作用提供重要见解： 通过阐明Cbfβ调节关节软骨中Wnt和雅普信号的机制， 再生和修复。为了实现研究目标，建立了一个多学科的研究团队。