Small integrin-binding proteins and tumor progression

小整合素结合蛋白与肿瘤进展

• 批准号: 7259482
• 负责人: NEAL S FEDARKO
• 金额: $ 25.09万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-15 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7259482
• 关键词: Animal Model; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biological Models; CD44 gene; Cell Culture System; Cell Surface Receptors; Cells; Chick Embryo; Chimera organism; Chromosomes, Human, Pair 4; Clinical Trials; Collagen; Complement Factor H; Data; Endothelial Cells; Enzyme Activation; Enzyme Inhibition; Exons; Failure; Family; Gel; Gene Cluster; Gene Family; Glycoproteins; Goals; Growth; Growth Factor; Host Defense; Human; In Vitro; Inhibition of Matrix Metalloproteinases Pathway; Integrin Binding; Kinetics; Knowledge; Ligands; Link; Malignant Neoplasms; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Mediating; Modeling; Molecular; Molecular Weight; Monitor; Mutate; Neoplasm Metastasis; Neoplasms; Nylons; Peptide antibodies; Physiological; Play; Polymerase Chain Reaction; Process; Proteins; RGD (sequence); Reagent; Recombinants; Role; Severity of illness; Siblings; Skeleton; Standards of Weights and Measures; System; Testing; Tissue Inhibitor of Metalloproteinases; Variant; Work; angiogenesis; base; cancer cell; cell motility; chorioallantoic membrane; implantation; in vivo; inhibitor/antagonist; member; neoplastic cell; protein expression; protein protein interaction; restoration; retinal rods; tumor growth; tumor progression

DESCRIPTION (provided by applicant): A family of secreted proteins, which we term SIBLINGS (for Small Integrin-Binding LIgand, N-linked Glycoproteins), contain the integrin-binding tripeptide, ROD, are expressed in the skeleton and have genes clustered on human chromosome 4. Our work and that of others has shown that a variety of neoplasms frequently express one or more of the proteins and that expression correlates with disease severity. We have recently found that SIBLINGS can bind to and modulate the activity of matrix metalloproteinases (MMPs). SIBLING binding to latent pro-MMPs leads to catalytic activity, and SIBLING binding to MMPs inhibited by tissue inhibitors of metalloproteinases (TIMPs) or small molecular weight inhibitors leads to a restoration of enzymatic activity. We hypothesize that SIBLING expression by neoplasms promotes tumor progression through MMP modulation. To test this hypothesis, we will (a) characterize SIBLING and MMP interactions using purified components and standard sequence and kinetic analyses to characterize SIBLING effects of MMP and MMP inhibitor kinetics as well as identify SIBLING modifying reagents; (b) test SIBLING- modifying reagents (variants, blocking peptides, antibodies) in in vitro cell systems that yield a functional readout on angiogenesis (endothelial cells and tubule formation) and invasiveness (human cancer cells and modified Boyden chamber assays); and (c) employ SIBLINGS, SIBLING-modifying reagents to demonstrate the physiological relevance of SIBLING - MMP interactions in tumor progression (using two quantitative chick embryo chorioallantoic membrane (CAM) systems to study angiogenesis and metastasis). The CAM angiogenesis assay employs the implantation of SIBLING reagents in a collagen gel within a nylon mesh and quantifying vessel formation. The CAM metastasis assay uses a highly sensitive assay based on PCR amplification of human alu sequences for monitoring the metastatic dissemination of human tumor cells in the chick embryo. The molecular action of SIBLINGS in modulating MMPs may contribute to tumor progression. The alteration of MMP activity by SIBLINGS and their expression in human cancer may be a contributory factor to the failure of MMP inhibitors in clinical trials. It is the long term goal of this work to exploit knowledge of the basic biochemical and biological details involved in SIBLING binding to MMP to develop anti-tumor growth and progression therapy based on disrupting or altering SIBLING - MMP interactions.

描述（由申请人提供）：一个分泌蛋白质的家族，我们称其为兄弟姐妹（对于小的整联蛋白结合配体，n连接的糖蛋白），包含内成蛋白结合的三肽，杆状杆菌，杆子，在骨架中表达，并在人类的工作中表达了更多的neoplass and ne neoplasm，并在s骨上表达了更多的表达。疾病的严重程度。我们最近发现，兄弟姐妹可以与基质金属蛋白酶（MMP）的活性结合并调节。兄弟姐妹与潜在pro-MMP的结合导致催化活性，而兄弟姐妹与MMP的结合被金属蛋白酶（TIMP）的组织抑制剂（TIMP）或小分子量抑制剂抑制，导致酶活性的恢复。我们假设肿瘤的同胞表达通过MMP调节促进肿瘤进展。为了检验这一假设，我们将（a）使用纯化的成分以及标准序列和动力学分析来表征同胞和MMP相互作用，以表征MMP和MMP抑制剂动力学的同胞效应，并识别兄弟姐妹修改的试剂； （b）在体外细胞系统中测试同胞修饰试剂（变体，阻断肽，抗体），这些试剂在血管生成（内皮细胞和小管形成）和侵入性（人类癌细胞和修饰的男孩室分析）上产生功能性读数； （c）采用兄弟姐妹修改试剂来证明肿瘤进展中同级MMP相互作用的生理相关性（使用两个定量的鸡胚胎绒毛膜绒毛膜（CAM）系统来研究血管生成和转移）。 CAM血管生成分析采用同胞试剂植入尼龙网格中的胶原蛋白凝胶并量化血管形成。 CAM转移测定法基于人类ALU序列的PCR扩增使用高度敏感的测定法，以监测雏鸡胚胎中人类肿瘤细胞的转移性传播。兄弟姐妹在调节MMP中的分子作用可能有助于肿瘤进展。兄弟姐妹对MMP活性的改变及其在人类癌症中的表达可能是临床试验中MMP抑制剂失败的一个因素。这项工作的长期目标是利用与MMP结合的基本生化和生物学细节知识，以基于破坏或改变兄弟姐妹 - MMP相互作用来发展抗肿瘤生长和进展治疗。