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

基本信息

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
项目类别：
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

项目摘要

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是我们实验室中鉴定出的细胞内LIM结构蛋白蛋白，该蛋白表明能够显着提高体外细胞对BMP-2的反应能力。该提案的长期目标是确认LMP-1通过调节对BMP-2信号很重要的蛋白酶体降解来调节对BMP的细胞反应性的假设。具体目的是证明LMP-1通过以下方面提高了对BMP-2的响应能力：1）中断R-SMADS的SMURF1介导的蛋白酶体降解（SMAD1）； 2）阻断SMURF1与I-SMADS（SMAD6）的相互作用，从而中断BMP受体（BMPR1A）的L-SMAD/SMURF1介导的蛋白酶体降解； 3）中断常见SMAD的JAB1介导的蛋白酶体降解（SMAD4）。我们的最终目标将确定LMP-1中的特定相互作用基序，从而使小分子的设计模仿LMP-1提高细胞对BMP-2的反应能力的能力。所采用的方法将包括蛋白质隔离，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体内对异位骨形成的测试。相关性：骨科中最大的临床挑战之一是无法持续产生脊柱融合和骨缺损重建的骨骼。 iLiac Crest骨移植物的使用可能无法在多达45％的患者中获得固体骨融合，而高达25％的患者可能会在供体部位遭受慢性疼痛。从这些实验中汲取的信息将进一步了解细胞形态发生蛋白的细胞反应性，骨形成的关键调节剂，并有可能导致设计小分子的设计，这些分子可以极大地提高BMPS的效力，从而使它们对患者的临床负担得起。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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.

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.

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