Altered elastogenesis by Tsk fibrillin-1 and in SSc

Tsk fibrillin-1 和 SSc 改变弹性生成

基本信息

批准号：
7667176
负责人：
ROBERT A. LAFYATIS
金额：
$ 19.81万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-20 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7667176
关键词：
Affect Attenuated Binding Binding Proteins Biology Breeding Cell Culture Techniques Cell Line Cells Clinical Collagen Data Deposition Dermal Dermis Elastic Fiber Elastin Employee Strikes Extracellular Matrix FBN1 Fibroblasts Fibrosis Functional disorder Genes Genotype Goals Human In Vitro Inflammatory Link Location Mediating Mediator of activation protein Messenger RNA Microfibrils Modeling Mus Pathogenesis Pathology Phenotype Proteins Regulation Research Personnel Role Signal Transduction Skeletal Muscle Skin Staining method Stains Study models Subcutaneous Tissue Systemic Scleroderma Testing Thick Tissues Transforming Growth Factor beta Transgenic Mice Tropoelastin cytokine fibrillin fibulin fibulin 2 human microfibrillar-associated protein 2 in vivo latent TGF-beta binding protein mRNA Expression microfibrillar protein mouse model mutant overexpression programs receptor skin disorder tsk mouse

项目摘要

DESCRIPTION (provided by applicant): A large duplication in the fibrillin-1 gene (Tsk fibrillin-1) is responsible for the phenotype of the Tsk mouse, a model for skin disease in human systemic sclerosis (SSc). The best defined function of fibrillin is in supporting elastic fibers and we have recently found that Tsk mice show markedly increased hypodermal elastic fibers associated with striking changes in fibulin-2 and fibulin-5, proteins that regulate formation of elastic fibers. We are investigating the hypotheses that altered hypodermal deposition of fibulins causes skin tethering in Tsk mice and that altered dermal fibulin deposition increases elastic fibers in human SSc. We will study the effect of Tsk fibrillin-1 on fibulin-2 and -5 matrices by staining cells conditionally expressing Tsk-fibrillin-1. We will test the importance of fibulin-2 on Tsk skin tethering and altered elastogenesis by breeding Tsk mice to fibulin-2 deleted mice, we will also study the effect of fibulin-2 on elastogenesis in fibroblasts overexpressing or deleted of fibulin-2 in cell culture. Fibrillin associates with several other matrix proteins, including MAGP-2. We have found markedly increased MAGP-2 matrix in the skin of Tsk mice and human SSc. Upon expression of Tsk fibrillin-1 in cell lines, we find increased MAGP-2 and collagen matrices, suggesting that Tsk fibrillin-1 might augment collagen deposition through increased MAGP-2. The effect of increased MAGP-2 expression on collagen, fibulin and elastic fiber matrices will be studied in cell lines and transgenic mice. TGF-beta may interact indirectly with fibrillin-1 through latent TGF-beta binding proteins and TGF-beta regulates fibrillin-1 and MAGP-2 matrices in vitro. Since these are features of Tsk dermal pathology, the importance of TGF-beta on the Tsk phenotype and altered elastogenesis will be tested by examining MAGP-2 and fibulin-2 expresssion in Tsk mice bred to smadS deleted or fibroblast-targeted TGF-beta Rll deleted mice. Fibroblast cultures from these mice will also be examined for alterations in TGF-beta activity, and MAGP-2 and fibulin-2 mRNA expression. LTBP-1 or LTBP-4 will be examined as possible mediators of Tsk fibrillin-1 activation of TGF-beta signaling. Together these studies should help clarify the pathogenesis of altered elastogenesis and fibrosis associated with Tsk fibrillin-1. These findings will be correlated with studies on human SSc skin.

描述（由申请人提供）：Fibrillin-1基因（TSK Fibrillin-1）中的大重复是TSK小鼠的表型，TSK小鼠是人类系统性硬化症（SSC）的皮肤病模型。原纤维蛋白的最佳定义功能是支持弹性纤维，我们最近发现，TSK小鼠显示出与纤维蛋白-2和纤维蛋白5的惊人变化相关的皮下注射弹性纤维，这是调节弹性纤维形成的蛋白质。我们正在研究改变腓蛋白皮下沉积的假设会导致TSK小鼠的皮肤束缚，并改变皮肤腓蛋白沉积会增加人类SSC中的弹性纤维。我们将通过染色有条件表达TSK - 原纤维蛋白-1的细胞来研究TSK Fibrillin-1对菲林-2和-5矩阵的影响。我们将通过将TSK小鼠繁殖到fibulin-2删除小鼠对Fibulin-2对TSK肤色绑扎的重要性，并改变了弹性发生，我们还将研究Fibulin-2对成纤维细胞过表达或在细胞培养中删除的成纤维细胞中弹性生成的影响。纤维蛋白与其他几种基质蛋白相关，包括MAGP-2。我们发现在TSK小鼠和人类SSC的皮肤中，Magp-2基质显着增加。在细胞系中Tsk Fibrillin-1表达后，我们发现MAGP-2和胶原蛋白矩阵增加，这表明TSK Fibrillin-1可能通过增加的MAGP-2增强胶原蛋白沉积。 MAGP-2表达增加对胶原蛋白，斐杜蛋白和弹性纤维基质的影响将在细胞系和转基因小鼠中进行研究。 TGF-β可以通过潜在的TGF-β结合蛋白与原纤维蛋白-1间接相互作用，而TGF-β可以在体外调节Fibrillin-1和Magp-2矩阵。由于这些是TSK真皮病理学的特征，因此TGF-beta在TSK表型上的重要性和改变的弹性发生，将通过检查在smads删除或成纤维细胞触发的TGF-beta-beta rll deleted小鼠的TSK小鼠中测试Magp-2和fibulin-2表达。还将检查来自这些小鼠的成纤维细胞培养物，以改变TGF-β活性的改变，以及MAGP-2和Fibulin-2 mRNA表达。 LTBP-1或LTBP-4将被检查为TSK纤维蛋白-1激活TGF-β信号传导的介体。这些研究共同有助于阐明与TSK纤维蛋白-1相关的弹性发生和纤维化改变的发病机理。这些发现将与人类SSC皮肤的研究相关。