LMP-1 is a critical regulator of BMP signaling and BMP responsiveness

LMP-1 是 BMP 信号传导和 BMP 反应性的关键调节因子

• 批准号: 7798565
• 负责人: SCOTT D. BODEN
• 金额: $ 24.08万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7798565
• 关键词: Address; Affinity; BMPR1A gene; Binding; Binding Proteins; Biological Assay; Bone Morphogenetic Proteins; Bone Transplantation; COPS5 gene; Cell Culture Techniques; Cell Nucleus; Cell membrane; Cells; Chemicals; Clinical; Complementary DNA; Complex; Data; Defect; Dose; Drug Design; Environment; Family member; Gene Expression; Genes; Genetic Transcription; Goals; Grant; Healed; Human; Implant; In Vitro; Keratectomy, Subepithelial, Laser-Assisted; Knowledge; LIM Domain Protein; Laboratories; Lead; Learning; Length; Leukocytes; Light; Location; Mediating; Mesenchymal Stem Cells; Methods; Modeling; Names; Nude Rats; Orthopedics; Oryctolagus cuniculus; Osteogenesis; Patients; Phosphorylation; Physiological; Plasmids; Preparation; Principal Investigator; Protein Family; Proteins; Publishing; Rattus; Recombinants; Regulation; Reporting; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Series; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Smad Proteins; Smad protein; Smad4 Protein; Solid; Spinal Fusion; Techniques; Technology; Testing; Time; Transforming Growth Factor beta; Translations; Ubiquitin; Ubiquitination; Vertebral column; Western Blotting; base; bone; bone morphogenetic protein 2; bone morphogenetic protein receptors; cell growth regulation; chronic pain; collected works; cost; design; experience; fetal; healing; improved; in vitro testing; in vivo; member; mimetics; mineralization; novel; osteoblast differentiation; overexpression; prevent; programs; receptor; reconstruction; research study; small molecule

DESCRIPTION (provided by applicant): PROJECT SUMMARY: LMP-1 is an intracellular LIM domain protein identified in our laboratory which has demonstrated an ability to dramatically increase cellular responsiveness to BMP-2 in vitro. The long term goals of this proposal are to confirm the hypothesis that LMP-1 modulates cellular responsiveness to BMPs through regulation of proteasomal degradation of key molecules that are important for BMP-2 signaling. The specific aims are to demonstrate that LMP-1 increases responsiveness to BMP-2 by: 1) interrupting Smurf1-mediated proteasomal degradation of R-Smads (Smad1); 2) blocking interaction of Smurf1 with I-Smads (Smad6), interrupting l-Smad/Smurf1-mediated proteasomal degradation of the BMP receptor (BMPR1A); and, 3) interrupting Jab1-mediated proteasomal degradation of the common Smad (Smad4). Our final aim will identify the specific interacting motifs in LMP-1, enabling design of a small molecule to mimic the ability of LMP-1 to enhance cellular responsiveness to BMP-2. The methods employed will include protein isolation, in vitro transcription/translation, Western blotting, ELSA, ubiquitination assays, real-time RT-PCR, computational drug design techniques, in vitro testing of BMP-2 responsiveness, and in vivo testing of ectopic bone formation. RELEVANCE: One of the greatest clinical challenges in Orthopaedics is the inability to consistently generate bone for spinal fusion and bone defect reconstruction. Use of iliac crest bone graft may fail to achieve solid bony fusion in up to 45% of patients, and up to 25% may experience chronic pain at the donor site. The information learned from these experiments will further the understanding of the regulation of cellular responsiveness to bone morphogenetic proteins, the key regulators of bone formation, and potentially lead to the design of small molecules that could greatly enhance the potency of BMPs, thereby making them more clinically affordable for patients.

描述（由申请人提供）： 项目摘要：LMP-1 是我们实验室鉴定的一种细胞内 LIM 结构域蛋白，已证明其能够在体外显着提高细胞对 BMP-2 的反应性。该提案的长期目标是证实 LMP-1 通过调节对 BMP-2 信号传导重要的关键分子的蛋白酶体降解来调节细胞对 BMP 的反应性的假设。具体目的是证明 LMP-1 通过以下方式增加对 BMP-2 的反应性：1) 中断 Smurf1 介导的 R-Smads (Smad1) 蛋白酶体降解； 2) 阻断Smurf1与I-Smads (Smad6)的相互作用，中断l-Smad/Smurf1介导的BMP受体(BMPR1A)的蛋白酶体降解； 3) 中断 Jab1 介导的常见 Smad (Smad4) 的蛋白酶体降解。我们的最终目标是确定 LMP-1 中的特定相互作用基序，从而能够设计一种小分子来模拟 LMP-1 的能力，从而增强细胞对 BMP-2 的反应性。所采用的方法包括蛋白质分离、体外转录/翻译、蛋白质印迹、ELSA、泛素化测定、实时RT-PCR、计算药物设计技术、BMP-2反应性的体外测试以及异位骨形成的体内测试。相关性：骨科最大的临床挑战之一是无法持续生成用于脊柱融合和骨缺损重建的骨骼。使用髂嵴骨移植可能无法在多达 45% 的患者中实现牢固的骨融合，并且多达 25% 的患者可能会在供体部位经历慢性疼痛。从这些实验中获得的信息将进一步了解细胞对骨形态发生蛋白（骨形成的关键调节因子）反应性的调节，并有可能导致小分子的设计，从而大大增强 BMP 的效力，从而使它们在临床上更容易为患者所接受。

项目成果

A synthetic compound that potentiates bone morphogenetic protein-2-induced transdifferentiation of myoblasts into the osteoblastic phenotype.

• DOI: 10.1007/s11010-010-0664-6
• 发表时间: 2011-03
• 期刊: MOLECULAR AND CELLULAR BIOCHEMISTRY
• 影响因子: 4.3
• 作者: Kato, Satoshi;Sangadala, Sreedhara;Tomita, Katsuro;Titus, Louisa;Boden, Scott D.
• 通讯作者: Boden, Scott D.

Characterization of a unique motif in LIM mineralization protein-1 that interacts with jun activation-domain-binding protein 1.

• DOI: 10.1007/s11010-013-1823-3
• 发表时间: 2014-01
• 期刊: Molecular and cellular biochemistry
• 影响因子: 4.3
• 作者: Sangadala S;Yoshioka K;Enyo Y;Liu Y;Titus L;Boden SD
• 通讯作者: Boden SD

Development and optimization of a cell-based assay for the selection of synthetic compounds that potentiate bone morphogenetic protein-2 activity.

• DOI: 10.1002/cbf.1615
• 发表时间: 2009-12
• 期刊: CELL BIOCHEMISTRY AND FUNCTION
• 影响因子: 3.6
• 作者: Okada, Motohiro;Sangadala, Sreedhara;Liu, Yunshan;Yoshida, Munehito;Reddy, Boojala Vijay B.;Titus, Louisa;Boden, Scott D.
• 通讯作者: Boden, Scott D.