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）：生长板中软骨细胞增殖、存活和 ECM 产生需要基质细胞蛋白 CCN 家族的几个成员。然而，CCN 介导这些活动的机制尚不清楚。同样，虽然一些 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 蛋白在健康成人软骨和 IVD 中的作用，以及它们在骨关节炎 (OA) 和退行性椎间盘疾病 (DDD) 等退行性疾病中的作用，目前尚无治疗方法。本提案中的研究解决了这些基本问题。核心假设是，CCN 1、2 和 4 在关节软骨和髓核 (NP) 的软骨形成和出生后维持中具有重叠功能，并且是它们所必需的，并且它们部分通过调节 HIF1a 和 HIF2a 介导其作用，部分通过充当 ECM 蛋白的护卫/伴侣来介导其作用。该假设将在三个具体目标上进行检验。在目标 1 中，将确定 CCN 1、2 和 4 在生长板软骨形成和关节软骨中的作用。携带 Ccns 1 和 2 的 floxed 等位基因的小鼠已经产生，Ccn4-/- 小鼠也已产生，并将用于产前和产后切除 Ccn 基因。初步研究支持 CCN 1 和 2 在产后软骨中具有重叠功能的假设。该目标的其他研究建立在初步数据的基础上，初步数据表明 Ccn2 的缺失会导致 NFkB (RelA) 的核定位减少，并降低 HIF1a 的水平，这两者在软骨中都具有促生存功能。 CCN 通过激活 NFkB /RelA 诱导 HIF1a 和 HIF2a 的假设得到了检验。在目标 2 中，通过诱导 CCN 切除的小鼠来测试 CCN 在 IVD 髓核 (NP) 形成和维持中的作用。这一目标基于初步数据，表明 Ccn2 突变体在 NP 形成方面具有主要缺陷，并且这些缺陷在 Ccn1/2 双突变体中加剧。该目的的其他实验测试了这样的假设：CCN 部分通过维持 HIF1a 和 HIF2a 的水平（如生长板软骨中的情况）来介导其对 ECM 产生和细胞存活的影响。在目标 3 中，测试了 CCN 1 和 2 直接充当“伴侣”以促进 ECM 组装的能力。这种作用模式的先例来自于对其他分泌蛋白具有这种活性的观察，以及 CCN 蛋白含有此类伴侣特有的 CXXC 基序的发现。这些研究具有创新性，因为他们利用新的小鼠模型来测试 CCN 的两种新作用模式：NFkB 和 HIF 对细胞活力和基质产生的调节，以及通过类伴侣活性在 ECM 组装中的直接作用。拟议的研究意义重大，因为它有望首次证明 CCN 对于维持关节软骨和 IVD 至关重要。这些知识有可能带来治疗 OA 和 DDD 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

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

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