The Role of CLIC 1 And 4 in Tumor Growth And Metastasis

CLIC 1 和 4 在肿瘤生长和转移中的作用

• 批准号: 8456821
• 负责人: Irina Jilishitz
• 金额: $ 4.15万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8456821
• 关键词: Ablation; Address; Adhesives; Affect; Angiogenic Factor; Animals; Architecture; Basement membrane; Biological Assay; Blood Vessels; CLIC4 gene; Cell physiology; Chloride Ion; Chlorides; Collaborations; Defect; Deposition; Development; Embryo; Endothelial Cells; Endothelium; Exhibits; Functional disorder; Genes; Growth; Hypoxia; Immigration; Implant; In Vitro; Knock-out; Knockout Mice; Laboratories; Lewis Lung Carcinoma; Maintenance; Measures; Mediating; Melanoma Cell; Membrane; Molecular; Monitor; Mus; Neoplasm Metastasis; Nucleic Acid Regulatory Sequences; Pathologic Neovascularization; Pathway interactions; Perfusion; Pericytes; Permeability; Proteins; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Smooth Muscle; Stratum Basale; Structure; Testing; Tissues; Tumor Angiogenesis; Vascular Permeabilities; Wild Type Mouse; adherent junction; angiogenesis; cadherin 5; cancer therapy; cell motility; density; in vitro Assay; intercalation; knockout animal; migration; mouse model; mutant; neoplastic cell; prevent; public health relevance; recombinase; sphingosine 1-phosphate; tumor; tumor growth; tumorigenesis

DESCRIPTION (provided by applicant): Angiogenesis is critical for tumor growth and metastasis and thus targeting tumor angiogenesis is a recognized means of blocking tumor growth. Chloride Intracellular Channel (CLIC) proteins, CLIC1 and CLIC4, are newly discovered angiogenic regulators that have been implicated in tumorigenesis. This proposal will first focus on the function of Clic4, where preliminary findings show that Clic4 deficiency leads to increased tumor metastasis. We hypothesize this increased metastasis is due to dysfunctional endothelium caused by reduced vascular wall barrier functions in Clic4 mutants. We propose to implant tumor cells on mice with an endothelial specific ablation of Clic4 and monitor tumor vessels, growth and metastasis. Next, we will determine if Clic1 and Clic4, the endothelially-expressed Clic genes, are required for tumor angiogenesis. We have found that deletion of both Clic1 and Clic4 genes in mice leads to embryonic lethality due to severe vascular defects. Thus, we propose to remove Clic4 in endothelial cells using a tissue specific cre recombinase driven by the Vegfr-2 regulatory region. The endothelial specific loss of Clic4 (Clic4ECKO) will be carried out in a Clic1 null background to study combined Clic1 and Clic4 function in tumor endothelium. To address the role of Clic1 and Clic4 in tumor angiogenesis, tumors will be implanted onto the Clic1-/- ;Clic4ECKO animals and tumor growth and angiogenesis will be evaluated. When evaluating tumor vasculature, the focus will be on assessing the amount, integrity, and patency of tumor vessels using immunohistochemical analysis. The level of tumor hypoxia will be examined, as a measure of reduced tumor vascular perfusion. The effects of combined loss of Clic1/Clic4 in tumor endothelium on tumor metastasis will also be determined. We have recently discovered the CLICs function in the S1P signaling pathway, a known signaling cascade involved in endothelial cell function. This proposal will investigate if CLICs function through the S1P signaling pathway to regulate endothelial cell migration and vascular barrier function using cultured primary endothelial cells with reduced expression of either CLIC1 or CLIC4. Using vitro assays we will determine CLICs are required for S1P functions in migration, vascular permeability and endothelial junction assembly. Determining whether CLICs function in tumor angiogenesis and metastasis and elucidating the molecular mechanism that mediate their function will provide new targets for development of anti-cancer therapies that can target both tumor growth and metastasis.

描述（由申请人提供）：血管生成对于肿瘤生长和转移至关重要，因此靶向肿瘤血管生成是阻断肿瘤生长的公认方法。氯化物内通道（CLIC）蛋白Clic1和Clic4是与肿瘤发生有关的新发现的血管生成调节剂。该提案将首先关注CLIC4的功能，在该功能中，初步发现表明CLIC4缺乏会导致肿瘤转移增加。我们假设这种增加的转移是由于CLIC4突变体中血管壁垒功能降低引起的功能失调的内皮。我们建议用Clic4的内皮特异性消融和监测肿​​瘤血管，生长和转移在小鼠上植入肿瘤细胞。接下来，我们将确定肿瘤血管生成需要Clic1和Clic4（内皮表达的Clic基因）。我们发现，小鼠中Clic1和Clic4基因的缺失导致由于严重的血管缺陷而导致胚胎致死性。因此，我们建议使用由VEGFR-2调节区域驱动的组织特异性CRE重组酶去除内皮细胞中的CLIC4。 Clic4（Clic4ecko）的内皮特异性损失将在CLIC1 NULL背景中进行研究，以研究肿瘤内皮中的CLIC1和CLIC4功能。为了解决CLIC1和CLIC4在肿瘤血管生成中的作用，将评估肿瘤在Clic1 - / - ; Clic4ecko动物以及肿瘤生长和血管生成上的作用。在评估肿瘤脉管系统时，重点将放在使用免疫组织化学分析评估肿瘤血管的数量，完整性和通畅性。将检查肿瘤缺氧的水平，以衡量减少肿瘤血管灌注的量度。还将确定CLIC1/CLIC4在肿瘤内皮中的组合损失对肿瘤转移的影响。我们最近在S1P信号通路中发现了CLICS函数，这是一种已知的信号传导级联，涉及内皮细胞功能。该建议将使用S1P信号传导途径进行调查，以使用培养的原代内皮细胞调节内皮细胞迁移和血管屏障功能，而CLIC1或CLIC4的表达降低。使用体外测定，我们将确定S1P功能在迁移，血管渗透性和内皮连接组件中所必需的库。确定CLIC在肿瘤血管生成和转移中的功能以及阐明介导其功能的分子机制是否会为开发抗癌疗法的开发提供新的靶标，这些抗癌疗法可以靶向肿瘤生长和转移。