Targeting CXCR4 and microRNA as Therapy for Chondrosarcoma

靶向 CXCR4 和 microRNA 治疗软骨肉瘤

• 批准号: 8503036
• 负责人: RICHARD M TEREK
• 金额: $ 28万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-25 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8503036
• 关键词: AMD3100; Adult; Animal Model; Attenuated; Bone neoplasms; CXC Chemokines; Cell Culture Techniques; Cell Line; Cell model; Cells; Chondrosarcoma; Clinical Data; Combined Modality Therapy; Cytotoxic Chemotherapy; Data; Effectiveness; Fibrinogen; G-Protein-Coupled Receptors; GTP-Binding Protein Regulators; Genetic Techniques; Genetic Transcription; Goals; Growth; HIF1A gene; Hand; Hypoxia; In Vitro; Interleukin-8B Receptor; Interstitial Collagenase; Laboratories; Lead; Link; Luciferases; Lung; Malignant Bone Neoplasm; Malignant Neoplasms; Measurement; Mediating; Metalloproteases; Metastatic Neoplasm to the Lung; MicroRNAs; Mission; Modeling; Molecular; Molecular Target; Monitor; Mus; National Cancer Institute; Neoplasm Metastasis; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Physiology; Public Health; RGS Proteins; Reporter; Research; Signal Pathway; Signal Transduction; Solid; Subfamily lentivirinae; Survival Rate; Testing; Therapeutic; Tissues; Tumor Angiogenesis; Tumor Volume; Untranslated Regions; Up-Regulation; Vascular Endothelial Growth Factors; Work; Xenograft Model; angiogenesis; animal data; base; bioimaging; cancer therapy; chemokine receptor; chemotherapy; driving force; extracellular; in vivo; inhibitor/antagonist; innovation; knock-down; mouse model; novel; novel strategies; overexpression; pre-clinical; public health relevance; receptor; research study; sarcoma; therapeutic target; translational approach; tumor; tumor growth; tumor progression

DESCRIPTION (provided by applicant): Chondrosarcoma is the only primary bone cancer that lacks an effective systemic treatment. It is the most common primary bone tumor in adults and survival is only 10-25%, with most patients succumbing to lung metastases. However, the mechanisms of metastasis, a strong potential target for therapeutics, are unknown. The long-term goal is to identify molecular targets for treatment. CXCR4 expression is endogenously upregulated in chondrosarcoma and its signaling increases expression of pro-metastatic factors. Specific microRNAs are also overexpressed in chondrosarcoma, and miR-181a in particular functions as an oncomir by enhancing CXCR4 signaling. Both CXCR4 signaling and miR-181a expression are also increased by hypoxia. The objective of this application is to use both cellular and animal model approaches to determine how overexpression of miR-181a is linked to metastasis in chondrosarcoma, and determine its appropriateness as a therapeutic target. CXCR4 is a G-protein-coupled receptor. Regulator of G-protein signaling protein sixteen (RGS16) has an inhibitory effect on CXCR4 signaling. Overexpression of specific microRNAs in cancer can disrupt inhibitory pathways, thereby promoting tumor progression. The central hypothesis is that miR-181a overexpression enhances CXCR4 signaling by inhibiting RGS16 and potentially other inhibitory pathways. This novel hypothesis is based on preliminary data produced in the applicant's laboratory. The rationale for this project is that the current antagonists for CXCR4 are only partly effective, suggesting a two-pronged approach based on targeting microRNA as well as the receptor, may be more effective. In this application, miR-181a as a CXCR4- enhancing oncomiR will be systematically investigated by pursuing two specific aims: 1) Determine the mechanistic link between hypoxia and chondrosarcoma metastasis; 2) Evaluate pharmacologic, anti-microRNA, and combined therapy in a chondrosarcoma mouse model. In Aim #1, overexpression of miR-181a in chondro- sarcoma cells will be inhibited using a lentivirus based expression construct for anti-miR-181a which has been proven feasible in the applicant's hands. The effects of such inhibition on expression of RGS16 and CXCR4 signaling will be analyzed. 3'-UTR luciferase reporters will be used to validate RGS16 as a target of miR-181a. In Aim #2, CXCR4 inhibition with anti-miR-181a, the drug AMD3100, and both will be evaluated in a mouse chondrosarcoma model for their effects on tumor growth, angiogenesis, and metastasis. In vivo bioimaging will be used to monitor matrix metalproteinase activity and angiogenesis in the tumors. The contribution of the pro- posed research is expected to be a detailed understanding of how microRNA and CXCR4 signaling regulate expression of MMPs, VEGF, and chondrosarcoma metastasis. This project is innovative because it utilizes anti-microRNA to inhibit CXCR4 signaling which may cause a shift from cytotoxic chemotherapy to biologic based therapy. The proposed research is significant because inhibition of CXCR4 signaling will provide the first biologically targeted therapy for chondrosarcoma.

描述（由申请人提供）：软骨肉瘤是唯一缺乏有效全身治疗的原发性骨癌。它是成人中最常见的原发性骨肿瘤，存活率仅为10- 25%，大多数患者死于肺转移。然而，转移的机制，一个强大的潜在的治疗目标，是未知的。长期目标是确定治疗的分子靶点。软骨肉瘤中CXCR 4表达内源性上调，其信号传导增加促转移因子的表达。特定的microRNA也在软骨肉瘤中过表达，并且miR-181 a特别地通过增强CXCR 4信号传导作为oncomir起作用。CXCR 4信号传导和miR-181 a表达也因缺氧而增加。本申请的目的是使用两种蜂窝 和动物模型方法来确定miR-181 a的过表达如何与软骨肉瘤中的转移相关联，并确定其作为治疗靶点的适当性。CXCR 4是一种G蛋白偶联受体。G蛋白信号调节蛋白16（Regulator of G-protein signaling protein 16，RGS 16）对CXCR 4信号通路具有抑制作用。癌症中特定microRNA的过表达可以破坏抑制途径，从而促进肿瘤进展。核心假设是miR-181 a过表达通过抑制RGS 16和潜在的其他抑制途径增强CXCR 4信号传导。这一新的假设是基于申请人实验室产生的初步数据。该项目的基本原理是，目前CXCR 4的拮抗剂仅部分有效，这表明基于靶向microRNA和受体的双管齐下的方法可能更有效。在本申请中，miR-181 a作为CXCR 4增强oncomiR将通过追求两个特定目标进行系统研究：1）确定缺氧和软骨肉瘤转移之间的机制联系; 2）在软骨肉瘤小鼠模型中评估药理学，抗microRNA和联合治疗。在目标#1中，使用基于慢病毒的抗miR-181 a表达构建体抑制软骨肉瘤细胞中miR-181 a的过表达，这在申请人手中已被证明是可行的。将分析这种抑制对RGS 16和CXCR 4信号传导的表达的影响。3 '-UTR荧光素酶报告基因将用于验证RGS 16作为miR-181 a的靶标。在目标#2中，将在小鼠软骨肉瘤模型中评价用抗miR-181 a、药物AMD 3100和两者抑制CXCR 4对肿瘤生长、血管生成和转移的影响。体内生物成像将用于监测肿瘤中的基质金属蛋白酶活性和血管生成。这项研究的贡献有望详细了解microRNA和CXCR 4信号传导如何调节MMPs、VEGF和软骨肉瘤转移的表达。该项目具有创新性，因为它利用抗microRNA来抑制CXCR 4信号传导，这可能导致从细胞毒性化疗转向基于生物的治疗。这项研究意义重大，因为抑制CXCR 4信号传导将为软骨肉瘤提供第一种生物靶向治疗。