Mechanisms of Synovial Joint Formation

滑膜关节形成机制

• 批准号: 10004500
• 负责人: MOTOMI ENOMOTO-IWAMOTO
• 金额: $ 42.38万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004500
• 关键词: 3-Dimensional; AXIN2 protein; Acute; Address; Adolescent; Adult; Affect; Age; Appearance; Behavior; Biomedical Engineering; Cartilage; Cartilage injury; Cell Count; Cell Lineage; Cell Proliferation; Cell Size; Cells; Chondrocytes; Cilia; Columnar Cell; Data; Degenerative polyarthritis; Development; Developmental Biology; Diphtheria Toxin; Disease; Drug Modelings; Embryonic Development; Epiphysial cartilage; Evolution; Genetic; Growth; Hip region structure; Immunohistochemistry; Injury; Joint by Site; Joint repair; Joints; Knee; Knockout Mice; Lateral; Lead; Left; Life; Limb structure; Location; Maintenance; Maps; Mesenchymal; Modeling; Monitor; Mus; Natural regeneration; Neonatal; Operative Surgical Procedures; Pathway interactions; Performance; Pharmacology; Phenotype; Population; Predisposition; Process; Quality of life; Regulatory Pathway; Reporter; Role; Signal Transduction; Specific qualifier value; Structure; Surface; Synovial joint; Testing; Therapeutic; Thick; Time; Tissues; age related; articular cartilage; base; beta catenin; bone epiphysis; cartilage development; cartilaginous; drug testing; genetic manipulation; image reconstruction; insight; intercalation; joint formation; long bone; lubricin; novel; planar cell polarity; postnatal; prenatal; progenitor; prospective; public health relevance; repaired; skeletal; smoothened signaling pathway; tissue repair; tool; transcriptome sequencing; translational medicine

 DESCRIPTION (provided by applicant): Synovial joints are essential for skeletal function, and much is known about their structure, distinct tissues and susceptibility to congenital and acquired diseases, including osteoarthritis (OA). In contrast, little is known about their developmental biology. Were such information available, it could be used directly or together with bioengineering tools to create new joint repair strategies. This project started several years ago to fill these important gaps. In the limb, joint formation initiates with appearance of a mesenchymal cell interzone at each prospective joint site within the cartilaginous long bone anlagen. However, it had long remained unclear whether the interzone cells serve as a transient joint demarcation point, whether they actually produce joint tissues over time and if so, how they perform that key task. To address these questions, we genetically tracked interzone cells using Gdf5 and found that Gdf5-Cre/R26-lineage cells persisted and produced joint tissues over time, thus representing specialized progenitors with joint formation capacity. We found also that Wnt/β- catenin signaling was important for surface zone cells expressing Prg4/lubricin. Though novel, the data left much unanswered, including how the Gdf5-lineage cells produce different joint tissues, how articular cartilage grows and acquires its important cell columnar organization, whether joint progenitors persist postnatally, and what roles they could have in repair. To address these issues, we created new Gdf5-CreER and Prg4-CreER lines and used others including Axin 2-CreER. Our preliminary data now indicate that: (a) progenitors within the prenatal joint are sequentially specified for contribution to different tissues; (b) lateral expanson of incipient articular cartilage over the growing long bone epiphysis involves progenitors near the groove of Ranvier with an Axin 2-lineage; (c) articular cartilage maturation and columnar organization involve cell intercalation mechanisms rather than appositional growth as suggested by others; and (iv) joint progenitor performance and Prg4 expression are enhanced by Kartogenin. These data lead to our central hypothesis that joint formation is a modular process brought about by progenitors specified sequentially during embryogenesis, with Wnt/β-catenin representing a major regulatory pathway. We also hypothesize that postnatal articular cartilage growth and maturation require specific changes in chondrocyte cytostructure and phenotype, and that joint cells' repair capacity is amenable to pharmacological or genetic stimulation. Our Aims are: (1) To fate-map joint progenitors during prenatal development and postnatal growth; (2) To determine mechanisms by which articular cartilage thickens and attains its columnar organization; and (3) To assess the repair capacity of joint progenitors and their progenies and means to boost it pharmacologically and/or genetically. The project will produce fundamentally new insights into the mechanisms of joint formation and repair and will thus have considerable basic and translational medicine relevance and significance.

 描述(申请人提供)：滑膜关节对骨骼功能至关重要，对其结构、不同的组织以及对先天性和后天疾病的易感性，包括骨关节炎(OA)，已有很多了解。相比之下，人们对它们的发育生物学知之甚少。如果有这样的信息，它可以直接使用或与生物工程工具一起创建新的联合修复策略。这个项目开始了好几年。 以填补这些重要的空白。在肢体中，关节的形成始于软骨长骨胶原内每个预期关节部位的间质细胞间带的出现。然而，长期以来，人们一直不清楚带间细胞是否作为临时的关节分界点，它们是否真的随着时间的推移产生关节组织，如果是的话，它们如何执行这一关键任务。为了解决这些问题，我们使用Gdf5对区域间细胞进行了遗传学追踪，发现Gdf5-Cre/R26系细胞随着时间的推移持续存在并产生关节组织，从而代表具有关节形成能力的特异祖细胞。我们还发现Wnt/β-catenin信号对表达Prg4/Luusin的表面区细胞很重要。虽然这些数据是新的，但仍有许多未回答的问题，包括GDF5系细胞如何产生不同的关节组织，关节软骨如何生长并获得其重要的细胞柱状组织， 联合祖先是否会在出生后持续存在，以及他们在修复过程中可能扮演什么角色。为了解决这些问题，我们创造了新的Gdf5-Creer和Prg4-Creer品系，并使用了包括Axin 2-Creer在内的其他品系。我们的初步数据现在表明：(A)产前关节内的祖细胞被顺序地指定为对不同组织的贡献；(B)早期关节软骨在生长的长骨骨骺上的侧向扩展涉及Ranvier沟附近的祖细胞，具有轴突2谱系；(C)关节软骨的成熟和柱状组织涉及细胞插入机制，而不是其他人所建议的同位生长；以及(Iv)Kartogenin促进了关节祖细胞的表现和Prg4的表达。这些数据导致了我们的中心假设，即关节形成是一个由胚胎发育过程中顺序指定的祖细胞带来的模块化过程，Wnt/β-Catenin是一个主要的调控途径。我们还假设，出生后关节软骨的生长和成熟需要软骨细胞结构和表型的特定变化，关节细胞的修复能力受到药物或遗传刺激的影响。我们的目标是：(1)绘制关节祖细胞在出生前发育和出生后生长的命运图；(2)确定关节软骨增厚并获得其柱状组织的机制；以及(3)评估关节祖细胞及其后代的修复能力，以及从药物和/或遗传上促进关节软骨修复的方法。该项目将从根本上对关节形成和修复的机制产生新的见解，因此将具有相当大的基础和转化医学相关性和意义。

项目成果

Reduced EGFR signaling enhances cartilage destruction in a mouse osteoarthritis model.

• DOI: 10.1038/boneres.2014.15
• 发表时间: 2014
• 期刊: BONE RESEARCH
• 影响因子: 12.7
• 作者: Zhang, Xianrong;Zhu, Ji;Liu, Fei;Li, Yumei;Chandra, Abhishek;Levin, L. Scott;Beier, Frank;Enomoto-Iwamoto, Motomi;Qin, Ling
• 通讯作者: Qin, Ling

Cell origin, volume and arrangement are drivers of articular cartilage formation, morphogenesis and response to injury in mouse limbs.

• DOI: 10.1016/j.ydbio.2017.04.006
• 发表时间: 2017-06-01
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Decker RS;Um HB;Dyment NA;Cottingham N;Usami Y;Enomoto-Iwamoto M;Kronenberg MS;Maye P;Rowe DW;Koyama E;Pacifici M
• 通讯作者: Pacifici M

Roles of β-catenin signaling in phenotypic expression and proliferation of articular cartilage superficial zone cells.

• DOI: 10.1038/labinvest.2011.144
• 发表时间: 2011-12
• 期刊: Laboratory investigation; a journal of technical methods and pathology

Resident mesenchymal progenitors of articular cartilage.

• DOI: 10.1016/j.matbio.2014.08.015
• 发表时间: 2014-10
• 期刊: MATRIX BIOLOGY
• 影响因子: 6.9
• 作者: Candela, Maria Elena;Yasuhara, Rika;Iwamoto, Masahiro;Enomoto-Iwamoto, Motomi
• 通讯作者: Enomoto-Iwamoto, Motomi

Distribution of slow-cycling cells in epiphyseal cartilage and requirement of β-catenin signaling for their maintenance in growth plate.

慢循环细胞在附后软骨中的分布以及β-catenin信号在生长板中维持的需求。

• DOI: 10.1002/jor.22583
• 发表时间: 2014-05
• 期刊: JOURNAL OF ORTHOPAEDIC RESEARCH
• 影响因子: 2.8
• 作者: Candela, Maria Elena;Cantley, Leslie;Yasuaha, Rika;Iwamoto, Masahiro;Pacifici, Maurizio;Enomoto-Iwamoto, Motomi
• 通讯作者: Enomoto-Iwamoto, Motomi