Control of bone homeostasis by MT1-MMP signaling

MT1-MMP 信号传导控制骨稳态

基本信息

批准号：
9109326
负责人：
Steven B Abramson
金额：
$ 22.26万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9109326
关键词：
Adipose tissue Adult Affect Age Alleles Amino Acids Applications Grants Arthritis Aspartic Acid Binding Bone Development Bone Growth Bone Marrow Bone Marrow Transplantation Bone Tissue Bone remodeling Cartilage Catalytic Domain Cell membrane Cells Charge Cytoplasmic Tail Data Defect Disease Dwarfism Engineering Epiphysial cartilage Exploratory/Developmental Grant Extracellular Matrix Fatty acid glycerol esters Female Femur Generations Genes Genetic Genetic Models Hajdu-Cheney Syndrome Homeostasis Homo Human Integral Membrane Protein Intervention Introns Knockout Mice Knowledge Lead Lipodystrophy LoxP-flanked allele MAPK3 gene MMP14 gene Matrix Metalloproteinases Mediating Mesenchymal Stem Cells Modeling Molecular Mus Mutant Strains Mice Mutation Myelogenous Osteoblasts Osteoclasts Osteogenesis Osteopenia Osteoporosis Pathologic Processes Pathway interactions Peptide Hydrolases Phenotype Phosphotyrosine Physiological Processes Play Property Proteolysis Role Signal Transduction Skeletal Development Skeletal bone Staging Syndrome Transplantation Tyrosine articular cartilage base bone bone mass cell type extracellular high reward high risk in vivo innovation insight long bone male mutant novel osteoblast differentiation osteogenic postnatal progenitor public health relevance recombinase skeletal substantia spongiosa tibia

项目摘要

DESCRIPTION (provided by applicant): We propose to investigate a novel mechanism by which membrane-type 1 matrix metalloproteinase (MT1- MMP, MMP-14) controls bone formation. MT1-MMP, an extracellular matrix-degrading, transmembrane proteinase with a 20-amino acid cytoplasmic tail, plays key roles in postnatal bone formation. In humans mutation of MT1-MMP causes the multicentric osteolysis and arthritis disease, Winchester syndrome. In the mouse MT1-MMP deficiency causes dwarfism, osteopenia and severe arthritis. MT1-MMP deletion in bone marrow-derived mesenchymal progenitor cells (MSC) fully recapitulates this phenotype, showing that MT1- MMP controls osteogenic differentiation in MSC. It has been proposed that the phenotype of MT1-MMP-/- mice results from the lack of the proteolytic activity of MT1-MMP. However, mounting evidence shows a variety of proteolysis-independent signaling functions of MT1-MMP. The unique tyrosine residue (Y573) in the MT1-MMP cytoplasmic tail is fundamental for the control of intracellular signaling. We have shown that Y573 controls activation of the Ras-ERK1/2 pathway, and that Y573 substitution with aspartic acid (D), a negatively charged amino acid like phosphotyrosine, blocks this function without affecting MT1-MMP proteolytic activity. We have therefore generated a mutant mouse with the Y573D substitution in the cytoplasmic tail of MT1-MMP. Our extensive studies of the phenotype of this mouse have led us to the unexpected and surprising finding that, MT1-MMP Y573D mice have a phenotype opposite to that of MT1-MMP-/- mice. Both homo- and hemizygous MT1-MMP Y573D mice show increased bone mass, increased osteoblast differentiation from MSC and decreased osteoclast differentiation from myeloid precursors. Importantly, MT1-MMP Y573D expression dramatically upregulates Wnt signaling in MSC. This is a novel and unexpected function of MT1-MMP that can be exploited for the generation of innovative treatments for a variety of conditions that alter bone homeostasis. Therefore, we propose to study it by developing the following Specific Aims: Aim 1) To generate and characterize the phenotype of mice with conditional expression of MT1-MMP Y573D in bone. We have already generated a MT1-MMP Y573D mouse in which the MT1-MMP locus contains a floxed neo cassette. Mice hemizygous for the neo cassette are phenotypically normal. We will cross these mice with mice that express Cre recombinase in either MSC or committed osteoblast progenitors, or in myeloid precursors of osteoclasts. Deletion of the neo cassette will result in heterozygous MT1-MMP Y573D mice, which have the phenotype we described. We will then analyze their bone phenotype, osteoblast and osteoclast differentiation and Wnt signaling. Aim 2) To characterize the role of MT1-MMP signaling in bone development and remodeling. We hypothesize that the phenotype of MT1-MMP Y573D mice can be transferred to wt mice by bone marrow transplantation. We will therefore transplant bone marrow from wt or MT1-MMP Y573D mice into young or adult, wt and MT1-MMP Y573D mice, and analyze their long bones 8 weeks post transplant.

描述（由适用提供）：我们建议研究一种新机制，膜型1基质金属蛋白酶（MT1-MMP，MMP-14）控制骨形成。 MT1-MMP是一种细胞外基质降解，具有20个氨基酸细胞质尾巴的跨膜蛋白酶，在产后骨形成中起关键作用。在人类突变中，MT1-MMP引起多中心骨溶解和关节炎疾病，温彻斯特综合征。在小鼠中，MT1-MMP缺乏会导致矮人，骨质减少和严重关节炎。骨髓衍生的间充质祖细胞（MSC）中的MT1-MMP缺失完全概括了这种表型，表明MT1-MMP控制MSC中的成骨分化。已经提出，MT1-MMP - / - 小鼠的表型是由于缺乏MT1-MMP的蛋白水解活性而导致的。然而，越野证据显示了MT1-MMP的各种蛋白水解非依赖性信号传导功能。 MT1-MP细胞质尾部中的独特酪氨酸居住（Y573）对于控制细胞内信号传导至关重要。我们已经表明，Y573控制RAS-ERK1/2途径的激活，并且用天冬氨酸（D）取代了Y573（一种像磷酸酪氨酸这样的负电荷氨基酸）阻止了该功能而不会影响MT1-MMP蛋白水解活性。因此，我们已经在MT1-MMP的细胞质尾部产生了一种用Y573D取代的突变小鼠。我们对这只小鼠表型的广泛研究使我们意外而令人惊讶的发现，MT1-MMP Y573D小鼠具有与MT1-MMP - / - 小鼠相反的表型。同型和半合子MT1-MMP Y573D小鼠均显示骨骼质量增加，成骨细胞与MSC的分化增加，重要的是MT1-MMP Y573D表达显着上调MSC中的Wnt信号。这是MT1-MMP的新颖且出乎意料的功能，可以为改变骨骼稳态的各种疾病的创新处理而探索。因此，我们建议通过开发以下特定目的来研究它：目的1）以骨骼中MT1-MMP Y573D的条件表达产生和表征小鼠的表型。我们已经生成了MT1-MMP Y573D小鼠，其中MT1-MMP基因座包含一个flox的Neo Cassette。新盒式的小鼠半合子在表型上是正常的。我们将与在MSC或成骨细胞祖细胞或破骨细胞的骨髓前体中表达CRE重组酶的小鼠穿越这些小鼠。新盒式的缺失将导致杂合MT1-MMP Y573D小鼠，这些小鼠具有我们描述的表型。然后，我们将分析其骨表型，成骨细胞和破骨细胞分化和Wnt信号传导。目标2）表征MT1-MMP信号在骨发育和重塑中的作用。我们假设MT1-MMP Y573D小鼠的表型可以通过骨髓移植转移到WT小鼠中。因此，我们将从WT或MT1-MMP Y573D小鼠移植到年轻人或成人，WT和MT1-MMP Y573D小鼠中，并在移植后8周分析其长骨头。