Nell-1, A Cbfa 1 Downstream Target, In Bone Formation

Nell-1，Cbfa 1 下游目标，在骨形成中

• 批准号: 7839166
• 负责人: KANG TING
• 金额: $ 40.07万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7839166
• 关键词: Address; Animals; Antibodies; Applications Grants; Bacteriophage T7; Bacteriophages; Binding; Binding Proteins; Binding Sites; Biological Assay; Bone Growth; Bone Regeneration; Cells; Cephalic; Chondrocytes; Clinical; Co-Immunoprecipitations; Commit; Craniosynostosis; Cysteine-Rich Domain; Data; Development; Down-Regulation; Epidermal Growth Factor; Family; Financial compensation; Foundations; Funding; Genetic Transcription; Goals; Hour; Human; Human Resources; Hypertrophy; In Vitro; Infant; Integrin Binding; Integrins; Investigation; Joint structure of suture of skull; Ligands; Mass Spectrum Analysis; Membrane Proteins; Molecular; N Domain; N-terminal; Natural regeneration; Osteoblasts; Osteogenesis; Parents; Patients; Peptide Signal Sequences; Phage Display; Progress Reports; Protein Isoforms; Proteins; Proteomics; Public Health; Recovery; Regulation; Resolution; Resources; Response Elements; Surgical sutures; System; Techniques; Testing; Therapeutic; Thrombospondin 1; Time; Tissues; Transcript; Transcriptional Regulation; United States National Institutes of Health; Up-Regulation; Work; abstracting; acute myeloid leukemia 1 protein; base; bone; cDNA Library; chordin; cis acting element; in vivo; novel; osteoblast differentiation; osteochondral tissue; osteogenic; parent grant; preclinical study; premature; promoter; public health relevance; receptor; receptor binding; relating to nervous system; response; secretory protein; selective expression

DESCRIPTION (provided by applicant): Project Summery/Abstract This Competitive Revision Application is in response to Notice Number (NOT-OD-09-058) and Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. We previously identified osteoinductive molecule, long Nell-1 [LNell-1; a protein strongly expressed in neural tissue encoding epidermal growth factor (EGF)-like domain] from prematurely fusing sutures in craniosynostosis (CS) patients (Many of these studies were performed before identification of the short Nell-1, SNell-1 isoform. For clarification, we will use "LNell-1" in place of Nell-1 when we are certain that the long form is involved. Meanwhile, Nell-1 can refer to both LNell-1 and SNell-1). LNell-1 is a secretory protein with a signal peptide, an NH2-terminal thrombospondin-1 (TSP1-N)-like module, five chordin-like cysteine-rich (CR) domains, and EGF-like domains. The long term goal of this Competitive Revision and parent grant proposal is to understand the mechanisms behind excessive bone growth in CS patients, so that we can harness and channel the molecules responsible into clinical therapies to regenerate bone. Towards this end, we have worked continuously to better understand the molecular regulation and function of LNell-1 so as to build a strong scientific foundation for therapeutic LNell-1 application. Our progress report and preliminary data for the parent grant Competitive Renewal demonstrate that: 1) runt-related transcription factor 2 (Runx2) regulates LNell-1 through cis-acting element 2 (OSE2) response elements (REs); 2) LNell-1 increases osteoblastic differentiation and can partially compensate for Runx2 haploinsufficiency; and 3) Nell-1 deletion animals demonstrate impaired chondrocyte hypertrophy with delayed bone formation as well as decreased Runx2 and increased osterix expression. For this one-year Competitive Revision we propose to significantly accelerate the tempo of our current investigations on mechanistic regulation of Nell-1 by a concentrated focus on identification of LNell-1 receptors and/or binding proteins (R/BPs). To accomplish this objective, we have obtained preliminary T7 phage display and mass spectrometry (MS)-based proteomic data demonstrating LNell-1 binding to membranous and cytosolic proteins. These data have led to the hypothesis that Nell-1 exerts its function in osteochondral differentiation by serving as a ligand to cell specific and potentially novel receptors/binding proteins. In order to identify LNell-1 R/BPs, three aims representing a logical extension of the parent proposal will be undertaken: Aim A) Candidate receptor approach to examine interaction of LNell-1 with integrin family. Structurally, the TSP1-N-like domain of LNell-1 shares a high degree of similarity with TSP1-N domain where integrin binding sites reside. Aim B) LNell-1 R/BP identification using a T7 phage display system. Aim C) MS-based functional proteomics on the membranous protein fraction binding. This Competitive Revision will also stimulate the economy by enabling hiring of additional staff and increasing hours of current part-time staff. PUBLIC HEALTH RELEVANCE: RELEVANCE TO PUBLIC HEALTH STATEMENT Nell-1, a cause of excessive bone growth in craniosynostosis patients has shown significant promise as a potent bone forming agent in preclinical studies. A better understanding of how and why Nell-1 works to form bone can accelerate the development of clinical therapies to help patients grow better and faster bone.

描述（由申请人提供）：项目摘要/摘要本竞争性修订申请是对通知编号（NOT-OD-09-058）和通知标题的回应：NIH宣布竞争性修订申请的恢复法案资金可用性。 我们先前从颅缝早闭（CS）患者的过早融合缝线中鉴定出骨诱导分子，长Nell-1 [LNell-1;一种在神经组织中强烈表达的蛋白质，编码表皮生长因子（EGF）样结构域]（许多这些研究是在鉴定短Nell-1，SNell-1亚型之前进行的。为了澄清，当我们确定涉及长形式时，我们将使用“LNell-1”代替Nell-1。同时，Nell-1可以指LNell-1和SNell-1）。LNell-1是一种分泌性蛋白，具有信号肽、NH 2-末端血小板反应蛋白-1（TSP 1-N）样模块、五个富含半胱氨酸（CR）的结构域和EGF样结构域。这项竞争性修订和母基金提案的长期目标是了解CS患者骨过度生长背后的机制，以便我们能够利用和引导负责临床治疗的分子来再生骨。为此，我们不断努力，以更好地了解LNell-1的分子调控和功能，从而为LNell-1的治疗应用奠定坚实的科学基础。我们的进展报告和竞争性更新的初步数据表明：1）runx相关转录因子2（Runx 2）通过顺式作用元件2（OSE 2）反应元件（RE）调节LNell-1：2）LNell-1促进成骨细胞分化并部分补偿Runx 2单倍不足;和3）Nell-1缺失动物显示软骨细胞肥大受损，骨形成延迟以及Runx 2降低和osterix表达增加。对于这个为期一年的竞争性修订，我们建议通过集中精力鉴定LNell-1受体和/或结合蛋白（R/BPs），显着加快我们目前对Nell-1机制调节的研究克里思。为了实现这一目标，我们已经获得了初步的T7噬菌体展示和质谱（MS）为基础的蛋白质组数据，证明LNell-1结合膜和胞质蛋白。这些数据导致了Nell-1通过作为细胞特异性和潜在的新型受体/结合蛋白的配体在骨软骨细胞分化中发挥其功能的假设。为了鉴定LNell-1 R/BP，将进行代表母提案的逻辑延伸的三个目标：目标A）检测LNell-1与整联蛋白家族的相互作用的候选受体方法。在结构上，LNell-1的TSP 1-N样结构域与整合素结合位点所在的TSP 1-N结构域具有高度的相似性。目的B）利用T7噬菌体展示系统鉴定LNell-1 R/BP。目的C）基于质谱的膜蛋白组分结合功能蛋白质组学研究。这一竞争性修订还将通过雇用更多的工作人员和增加目前兼职工作人员的工作时间来刺激经济。 公共卫生关系：与公共卫生声明的相关性Nell-1是颅缝早闭患者骨过度生长的一个原因，在临床前研究中显示出作为有效骨形成剂的重大前景。更好地了解Nell-1如何以及为什么形成骨骼可以加速临床治疗的发展，以帮助患者更好更快地生长骨骼。

项目成果

Preparation and characterization of trace elements-multidoped injectable biomimetic materials for minimally invasive treatment of osteoporotic bone trauma.

• DOI: 10.1002/jbm.a.32936
• 发表时间: 2010-12
• 期刊: Journal of biomedical materials research. Part A
• 作者: Xianyan Yang;Yilai Gan;Xin Gao;Li Zhao;Changyou Gao;Xinli Zhang;Yanbo Feng;K. Ting;Z. Gou

The use of BMP-2 coupled - Nanosilver-PLGA composite grafts to induce bone repair in grossly infected segmental defects.

BMP-2耦合的使用 - 纳米层-PLGA复合移植物在严重感染的节段缺陷中诱导骨修复。

• DOI: 10.1016/j.biomaterials.2010.08.041
• 发表时间: 2010-12
• 期刊: BIOMATERIALS
• 影响因子: 14
• 作者: Zheng, Zhong;Yin, Wei;Zara, Janette N.;Li, Weiming;Kwak, Jinny;Mamidi, Rachna;Lee, Min;Siu, Ronald K.;Ngo, Richard;Wang, Joyce;Carpenter, Doug;Zhang, Xinli;Wu, Benjamin;Ting, Kang;Soo, Chia
• 通讯作者: Soo, Chia

A new function of Nell-1 protein in repressing adipogenic differentiation.

• DOI: 10.1016/j.bbrc.2011.06.111
• 发表时间: 2011-07-22
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: James, Aaron W.;Pan, Angel;Chiang, Michael;Zara, Janette N.;Zhang, Xinli;Ting, Kang;Soo, Chia
• 通讯作者: Soo, Chia

The role of NELL-1, a growth factor associated with craniosynostosis, in promoting bone regeneration.

• DOI: 10.1177/0022034510376401
• 发表时间: 2010-09
• 期刊: Journal of dental research
• 影响因子: 7.6
• 作者: Zhang X;Zara J;Siu RK;Ting K;Soo C
• 通讯作者: Soo C

NELL-1 binds to APR3 affecting human osteoblast proliferation and differentiation.

• DOI: 10.1016/j.febslet.2011.06.024
• 发表时间: 2011-08-04
• 期刊: FEBS letters
• 影响因子: 3.5
• 作者: Zou X;Shen J;Chen F;Ting K;Zheng Z;Pang S;Zara JN;Adams JS;Soo C;Zhang X
• 通讯作者: Zhang X