CCN PROTEINS IN CARTILAGE FORMATION AND MAINTENANCE

CCN 蛋白在软骨形成和维护中的作用

• 批准号: 8890646
• 负责人: Karen M. Lyons
• 金额: $ 41.18万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-20 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8890646
• 关键词: Address; Adult; Arthritis; Biological Assay; Cartilage; Cell Survival; Cells; Characteristics; Chondrocytes; Chondrogenesis; Circular Dichroism; Data; Defect; Degenerative Disorder; Degenerative polyarthritis; Development; Disease; Epiphysial cartilage; Excision; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Family; Family member; Genes; Hypoxia; In Vitro; Intervertebral disc structure; Knowledge; Lead; Low Back Pain; LoxP-flanked allele; Maintenance; Mediating; Mediator of activation protein; Molecular Chaperones; Mus; Nuclear; Organ Culture Techniques; Outcome; Pathway interactions; Play; Production; Protein Disulfide Isomerase; Proteins; Publications; Regulation; Research; Role; Signal Transduction; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Up-Regulation; WISP1 gene; Work; articular cartilage; base; chondrodysplasia; disability; effective therapy; endoplasmic reticulum stress; hypoxia inducible factor 1; innovation; intervertebral disk degeneration; member; mouse model; mutant; novel; novel therapeutic intervention; nucleus pulposus; overexpression; postnatal; prevent; research study; response; skeletal tissue; targeted treatment; therapy development

DESCRIPTION (provided by applicant): Several members of the CCN family of matricellular proteins are required for chondrocyte proliferation, survival, and ECM production in the growth plate. However, the mechanisms by which CCNs mediate these activities are unknown. Similarly, although some CCN proteins are expressed in adult articular cartilage and intervertebral discs (IVDs), their functions in these tissues are unknown. Thus there is a large gap in knowledge regarding the roles of CCN proteins in healthy adult cartilage and IVDs, and their roles in degenerative diseases such as osteoarthritis (OA) and degenerative disc disease (DDD), for which there are currently no therapies. The studies in this proposal address these fundamental issues. The central hypothesis is that CCNs 1, 2, and 4 have overlapping functions in and are required for chondrogenesis and postnatal maintenance of articular cartilage and nucleus pulposus (NP), and that they mediate their effects in part through regulation of HIF1a and HIF2a, and in part by acting as escorts/chaperones for ECM proteins. This hypothesis will be tested in three specific aims. In Aim 1, the roles of CCNs 1, 2, and 4 in growth plate chondrogenesis and articular cartilage will be determined. Mice carrying floxed alleles of Ccns 1 and 2 have been generated, as have Ccn4-/- mice, and will be used to excise Ccn genes prenatally and postnatally. Preliminary studies support the hypothesis that CCNs 1 and 2 have overlapping functions in postnatal cartilage. Other studies in this aim build on preliminary data showing that loss of Ccn2 leads to decreased nuclear localization of NFkB (RelA), and decreased levels of HIF1a, both of which have pro-survival functions in cartilage. The hypothesis that CCNs induce HIF1a and HIF2a by activating NFkB /RelA is tested. In Aim 2, the roles of CCNs in the formation and maintenance of the nucleus pulposus (NP) of the IVD are tested by generating mice in which excision of CCNs is induced. This aim is based on preliminary data showing that Ccn2 mutants have primary defects in formation of the NP, and these are exacerbated in Ccn1/2 double mutants. Other experiments in this aim test the hypothesis that CCNs mediate their effects on ECM production and cell survival in part through maintaining levels of HIF1a and HIF2a, as in growth plate cartilage. In Aim 3, the ability of CCNs 1 and 2 to act directly as "chaperones" to facilitate ECM assembly is tested. Precedent for this mode of action comes from observations that other secreted proteins have this activity, and from the finding that CCN proteins contain CXXC motifs characteristic of such chaperones. The studies are innovative, because they utilize new mouse models to test two novel modes of action for CCNs: regulation of cell viability and matrix production by NFkB and HIFs, and direct roles in ECM assembly via chaperone-like activity. The proposed research is significant, because it is expected to demonstrate for the first time that CCNs are essential for the maintenance of articular cartilage and IVD. This knowledge has the potential to lead to new therapeutic approaches to the treatment of OA and DDD.

描述（由申请人提供）：生长板中的软骨细胞增殖，生存和ECM产生需要CCN的几个成员。但是，CCN介导这些活动的机制尚不清楚。同样，尽管某些CCN蛋白在成人关节软骨和椎间盘（IVD）中表达，但它们在这些组织中的功能尚不清楚。因此，关于CCN蛋白在健康的成人软骨和IVD中的作用的知识差距很大，及其在退行性疾病中的作用，例如骨关节炎（OA）（OA）和退行性椎间盘疾病（DDD），目前尚无治疗疗法。该提案中的研究解决了这些基本问题。中心假设是CCNS 1、2和4具有重叠的功能，并且需要对关节软骨和核核（NP）（NP）的出生和产后维持（NP）的维护，并且它们通过调节HIF1A和HIF2A的调节，部分地介导了它们的作用。该假设将以三个特定目的进行检验。在AIM 1中，将确定CCNS 1、2和4在生长板软骨发生和关节软骨中的作用。与CCN4 - / - 小鼠一起产生了携带CCN 1和2等位基因的小鼠，并将用于产前和产后对CCN基因进行消费。初步研究支持以下假设：CCNS 1和2在产后软骨中具有重叠的功能。此目标的其他研究基于初步数据，表明CCN2的损失导致NFKB（RELA）的核定位降低，而HIF1A的水平降低，这两者在软骨中都具有亲生性功能。测试了CCN通过激活NFKB /RELA诱导HIF1A和HIF2A的假设。在AIM 2中，通过产生诱导CCNS切除的小鼠来测试CCN在IVD核（NP）的形成和维持中的作用。该目的基于初步数据，表明CCN2突变体在形成NP时具有主要缺陷，并且在CCN1/2双突变体中会加剧这些缺陷。此目标中的其他实验检验了CCNS在维持HIF1A和HIF2A的水平（如生长板软骨中）介导其对ECM产生和细胞存活的影响的假设。在AIM 3中，测试了CCN 1和2直接充当“伴侣”促进ECM组装的能力。这种作用方式的先例来自于观察到其他分泌蛋白具有这种活性，并且从发现CCN蛋白包含此类伴侣特征的CCN蛋白的发现。这些研究具有创新性，因为它们利用新的小鼠模型来测试CCN的两种新型作用模式：NFKB和HIF对细胞活力和基质产生的调节，并通过类似伴侣的活性在ECM组装中直接作用。拟议的研究很重要，因为预计将首次证明CCN对于维持关节软骨和IVD至关重要。这些知识有可能导致OA和DDD治疗的新治疗方法。

项目成果

CCN family 2/connective tissue growth factor modulates BMP signalling as a signal conductor, which action regulates the proliferation and differentiation of chondrocytes.

• DOI: 10.1093/jb/mvn159
• 发表时间: 2009-02
• 期刊: Journal of biochemistry
• 影响因子: 2.7
• 作者: Maeda A;Nishida T;Aoyama E;Kubota S;Lyons KM;Kuboki T;Takigawa M
• 通讯作者: Takigawa M

CCN family 2/connective tissue growth factor (CCN2/CTGF) regulates the expression of Vegf through Hif-1alpha expression in a chondrocytic cell line, HCS-2/8, under hypoxic condition.

• DOI: 10.1016/j.bone.2008.08.125
• 发表时间: 2009-01
• 期刊: BONE
• 影响因子: 4.1
• 作者: Nishida, Takashi;Kondo, Seiji;Maeda, Azusa;Kubota, Satoshi;Lyons, Karen M.;Takigawa, Masaharu
• 通讯作者: Takigawa, Masaharu

CCN family 2/connective tissue growth factor (CCN2/CTGF) promotes osteoclastogenesis via induction of and interaction with dendritic cell-specific transmembrane protein (DC-STAMP).

• DOI: 10.1002/jbmr.222
• 发表时间: 2011-02
• 期刊: JOURNAL OF BONE AND MINERAL RESEARCH
• 影响因子: 6.2
• 作者: Nishida, Takashi;Emura, Kenji;Kubota, Satoshi;Lyons, Karen M.;Takigawa, Masaharu
• 通讯作者: Takigawa, Masaharu

CCN2/CTGF is required for matrix organization and to protect growth plate chondrocytes from cellular stress.

• DOI: 10.1007/s12079-013-0201-y
• 发表时间: 2013-08
• 期刊: JOURNAL OF CELL COMMUNICATION AND SIGNALING
• 影响因子: 4.1
• 作者: Hall-Glenn, Faith;Aivazi, Armen;Akopyan, Lusi;Ong, Jessica R.;Baxter, Ruth R.;Benya, Paul D.;Goldschmeding, Roel;van Nieuwenhoven, Frans A.;Hunziker, Ernst B.;Lyons, Karen M.
• 通讯作者: Lyons, Karen M.

CCN2 as a novel molecule supporting energy metabolism of chondrocytes.

• DOI: 10.1002/jcb.24728
• 发表时间: 2014-05
• 期刊: JOURNAL OF CELLULAR BIOCHEMISTRY
• 影响因子: 4
• 作者: Maeda-Uematsu, Aya;Kubota, Satoshi;Kawaki, Harumi;Kawata, Kazumi;Miyake, Yoshiaki;Hattori, Takako;Nishida, Takashi;Moritani, Norifumi;Lyons, Karen M.;Iida, Seiji;Takigawa, Masaharu
• 通讯作者: Takigawa, Masaharu