Human Copper Transporter 1 as Reporter Gene for Imaging

人类铜转运蛋白 1 作为成像报告基因

• 批准号: 7196211
• 负责人: FANGYU PENG
• 金额: $ 18.11万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7196211
• 关键词: Affinity; Animal Model; Bladder; Brain; Cells; Cervix carcinoma; Cisplatin; Clinical Trials; Copper; Data; Dependovirus; Development; Dopamine Receptor; Drug or chemical Tissue Distribution; Ensure; Experimental Animal Model; Gene Delivery; Genes; Half-Life; Head and Neck Cancer; Hela Cells; Hepatic; Hepatic Tissue; Herpesviridae; Hour; Human; Human body; Image; Imaging Techniques; Invasive; Liver; Malignant neoplasm of prostate; Malignant neoplasm of urinary bladder; Mediating; Monitor; Mus; Neuroblastoma; Oncogene Proteins; Oncogenes; Organ; Pattern; Phosphotransferases; Pilot Projects; Positron-Emission Tomography; Prostate; Purpose; Radioactive; Reaction; Reporter; Reporter Genes; Reporting; Research Personnel; Safety; Salivary Glands; Signal Transduction; Site; Tissues; Tracer; Tumor Tissue; Viral; Xenograft procedure; adeno-associated viral vector; base; chemotherapeutic agent; copper transporter 1; gene therapy; in vivo; innovation; manufacturing facility; molecular imaging; novel; programs; promoter; research study; size; sodium-iodide symporter; therapeutic gene; transgene expression; tumor; tumor xenograft; uptake; vector

DESCRIPTION (provided by applicant): It is essential to monitor transgene expression for efficacy and safety of human gene therapy. Use of a reporter gene to monitor gene delivery and expression in targeted tissue represents an innovative molecular imaging technique. After simultaneous delivery of a reporter gene with a therapeutic gene, imaging signals from a reporter gene indirectly reflect the distribution and expression of a therapeutic gene in vivo. Because there are some limitations associated with the reporter genes currently available, it is necessary to search for new reporter genes which may be used when use of other reporter genes is limited. Human copper transporter 1 (hCtrl) gene encodes a high affinity copper transporter. Endogenous hCtrl is highly expressed in liver, with much lower expression in extra-hepatic tissues. Here, we hypothesize that hCtrl may be used as a reporter gene for imaging gene delivery in vivo by PET. We will perform exploratory experiments with two specific aims. AIM 1: To determine whether over-expression of hCtrl in tumor tissue can be imaged non-invasively and quantitatively by Cu-64 PET, using human cervical carcinoma xenograft in mice as an experimental animal model. AIM 2: To evaluate feasibility using hCtrl as a reporter gene for imaging transcriptional targeting of hCtrl expression to tumor over-expressing N-MYC oncoprotein by Cu-64 PET, using human neuroblastoma xenograft in mice as an animal model. This new reporter gene may have unique desirable features in comparison to other reporter genes: 1) hCtrl will not stimulate immunological reaction in humans and is expected to be very useful for longitudinal monitoring of transgene expression non-invasively and quantitatively; 2) the hCtrl reporter gene is expected to be especially useful for imaging targeted gene delivery in prostate cancer gene therapy because there is little background activity in the urinary bladder; 3) hCtrl holds potential as a therapeutic gene based on its capability to medicate cellular uptake of cisplatin. The data from this pilot study will help us to determine whether it is feasible to use hCtrl as a reporter gene for imaging. Successful development of this new hCtrl reporter gene is expected to contribute to advance of human gene therapy significantly by providing a versatile reporter gene for longitudinal monitor of transgene expression in vivo non-invasively and qunatitatively by PET imaging.

描述（申请人提供）：监测转基因表达对人类基因治疗的有效性和安全性至关重要。使用报告基因来监测靶组织中的基因递送和表达代表了创新的分子成像技术。在报告基因与治疗基因同时递送后，来自报告基因的成像信号间接反映治疗基因在体内的分布和表达。由于目前可用的报道基因存在一些限制，因此有必要寻找新的报道基因，当其他报道基因的使用受到限制时，可以使用这些报道基因。人铜转运蛋白1（hCtrl）基因编码高亲和力铜转运蛋白。内源性hCtrl在肝脏中高表达，在肝外组织中表达低得多。在这里，我们假设hCtrl可以用作报告基因，用于通过PET在体内成像基因递送。我们将进行探索性实验，有两个具体目标。目标1：以人宫颈癌小鼠移植瘤为实验动物模型，探讨Cu-64 PET对肿瘤组织中hCtrl过度表达的无创性定量成像。目标2：以小鼠人神经母细胞瘤异种移植瘤为动物模型，评价以hCtrl为报告基因，通过Cu-64 PET成像hCtrl表达对过表达N-MYC癌蛋白的肿瘤的转录靶向的可行性。与其它报告基因相比，该新报告基因可能具有独特的理想特征：1）hCtrl不会刺激人体的免疫反应，并且预期对于非侵入性和定量地纵向监测转基因表达是非常有用的;（二）hCtrl报告基因预期特别适用于前列腺癌基因治疗中的靶向基因递送成像，因为在前列腺癌基因治疗中几乎没有背景活性。膀胱; 3）hCtrl基于其抑制顺铂的细胞摄取的能力而具有作为治疗基因的潜力。这项初步研究的数据将帮助我们确定是否可以使用hCtrl作为报告基因进行成像。这种新的hCtrl报告基因的成功开发，预计将有助于人类基因治疗的进步显着提供一个通用的报告基因的纵向监测转基因的表达在体内非侵入性和定量的PET成像。