Regulation of heparanase in cancer progression

乙酰肝素酶在癌症进展中的调节

基本信息

批准号：
7215163
负责人：
ISRAEL VLODAVSKY
金额：
$ 16.79万
依托单位：
TECHNION-ISRAEL INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2009-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7215163
关键词：
Address Affect Angiogenic Factor Animal Model Animals Antibodies Architecture Binding Biological Biological Assay Blood Vessels Cancer Patient Cancerous Catalytic RNA Cell Adhesion Cell surface Cloning Development Disease Endo-beta-D-Glucuronidase Enzymes Equilibrium Experimental Models Extracellular Matrix Gene Expression Gene Proteins Gene Silencing Generations Genes Genetic Transcription Heparan Sulfate Proteoglycan Heparitin Sulfate Human Invasive Knockout Mice Laboratories Longitudinal Studies Magnetic Resonance Imaging Mediating Membrane Methylation Molecular Probes Mutate Neoplasm Metastasis Nuclear Protein Nuclear Proteins Oligosaccharides Phenotype Physiology Play Postoperative Period Process Property Protein Overexpression Proteins Proteolytic Processing Recombinants Regulation Research Response Elements Role Small Interfering RNA System Testing Time Tissues Transgenic Mice Transgenic Organisms Tumor Angiogenesis Vascular Endothelial Growth Factors angiogenesis cancer therapy cell behavior cell motility density design drug development heparanase in vivo inhibitor/antagonist interest migration mouse heparanase neoplastic cell neutralizing antibody novel strategies promoter response self assembly stem steroid hormone tool tumor tumor growth tumor progression uptake

项目摘要

DESCRIPTION (provided by applicant): Heparan sulfate (HS) proteoglycans play a key role in the self-assembly, insolubility and barrier properties of the extracellular matrix (ECM). Cleavage of HS therefore affects the integrity of tissues and hence normal and pathological phenomena involving cell migration and response to changes in the ECM. Mammalian heparanase, endo-beta-D-glucuronidase, first cloned and characterized in our laboratory, is synthesized as a latent 65 kDa protein that is processed into a highly active 50 kDa enzyme. Heparanase is preferentially expressed in human tumors and its overexpression in tumor cells confers an invasive phenotype in experimental animals. Heparanase also releases angiogenic factors from the ECM and thereby induces an angiogenic response in vivo. Enhanced heparanase expression correlates with metastatic potential, tumor vascularity and reduced postoperative survival of cancer patients. These observations, the anti-cancerous effect of heparanase-inhibiting molecules, and the unexpected identification of a predominant functional heparanase suggest that the enzyme is a promising target for drug development. Given the potential tissue damage that could result from inadvertent cleavage of HS, tight regulation is essential. Of particular interest is the significance of cell surface expression and secretion of heparanase, its proteolytic processing, cellular uptake, and effects on cell adhesion, migration, metastasis, and angiogenesis. We propose to investigate the involvement of heparanase in cancer progression, emphasizing regulatory aspects, normal functions, and non-enzymatic activities of the molecule, as well as approaches to efficiently inhibit heparanase expression and activity. Specifically, we will (I) investigate the regulation of heparanase promoter activity and characterize the proteolytic activity and molecular interactions involved in processing and activation of latent heparanase; (II) generate and characterize heparanase transgenic and knockout mice and elucidate heparanase normal functions and interaction with cellular proteins; (III) study involvement of heparanase, expressed by the tumor and its microenvironment, in tumor growth, angiogenesis, and metastasis; and (IV) develop inhibitors of heparanase gene expression and enzymatic activity. Efficient inhibitors will be tested in experimental models of tumor progression. The proposed research stems from studies performed since the cloning of the heparanase gene, and availability of molecular probes (i.e., recombinant latent and active heparanases; intracellular, secreted and point mutated species of heparanase, anti-heparanase antibodies), assay systems (i.e., MRI analysis of vascular density and functionality) and animal models to elucidate causal involvement of heparanase in cancer progression. It is through understanding of the enzyme's physiology and regulation that adequate heparanase-inhibiting strategies can be designed and applied in the treatment of cancer and possibly other disorders involving HS and heparanase.

描述（由申请人提供）：硫酸乙酰肝素（HS）蛋白聚糖在自组装，不溶性和屏障特性（ECM）中起关键作用。因此，HS的切割会影响组织的完整性，因此涉及细胞迁移以及对ECM变化的反应的正常和病理现象。哺乳动物乙酰肝素酶，贝塔-Beta-D-葡萄糖醛酸苷酶首先在我们的实验室中进行了特征，被合成为潜在的65 kDa蛋白，该蛋白被加工成高度活跃的50 kDa酶。肝素酶在人类肿瘤中优先表达，其在肿瘤细胞中的过表达赋予了实验动物中的侵入性表型。乙酰肝素酶还从ECM释放血管生成因子，从而在体内诱导血管生成反应。增强的肝素酶表达与转移性潜力，肿瘤血管性和癌症患者术后存活降低相关。这些观察结果，抗肝素酶抑制分子的抗癌作用以及对主要功能性肝素酶的意外鉴定表明该酶是药物发育的有希望的靶标。鉴于HS无意裂解可能导致的潜在组织损伤，因此严格的调节是必不可少的。特别感兴趣的是细胞表面表达和肝素分泌的重要性，其蛋白水解加工，细胞摄取以及对细胞粘附，迁移，转移和血管生成的影响。我们建议研究肝素酶参与癌症进展，强调分子的调节方面，正常功能和非酶活性，以及有效抑制肝素酶表达和活性的方法。具体而言，我们将（i）研究肝素酶启动子活性的调节，并表征参与潜在乙酰肝素酶的加工和激活所涉及的蛋白水解活性和分子相互作用；（ii）生成和表征肝素酶转基因和基因敲除小鼠，并阐明肝素酶正常功能以及与细胞蛋白的相互作用；（iii）研究的研究涉及肝素酶，由肿瘤及其微环境表达，参与肿瘤生长，血管生成和转移；（iv）发展肝素酶基因表达和酶促活性的抑制剂。有效的抑制剂将在肿瘤进展的实验模型中进行测试。拟议的研究源于自肝素酶基因克隆以来进行的研究，以及分子探针的可用性（即重组潜伏和活性乙酰肝素酶；细胞内，分泌和点突变的乙酰肝素酶，抗肝酶抗体），分析系统（即，测定系统（，测定）对血管密度和功能的MRI分析）和动物模型，以阐明肝素酶在癌症进展中的因果关系。正是通过了解酶的生理学和调节，可以设计和应用适当的肝酶抑制策略，以治疗癌症以及可能涉及HS和肝素酶的其他疾病。