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: 7348310
• 负责人: FANGYU PENG
• 金额: $ 4.71万
• 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 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7348310
• 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（由申请人提供）：监测转基因表达对于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目标组织中的基因传递和表达代表了一种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。由于目前可用的报告基因存在一些局限性，因此有必要寻找新的报告基因，以便在其他报告基因的使用受到限制时可以使用。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 可能用作报告基因，通过 PET 对体内基因传递进行成像。我们将进行具有两个具体目标的探索性实验。目的1：以小鼠人宫颈癌异种移植物作为实验动物模型，确定是否可以通过Cu-64 PET对肿瘤组织中hCtrl的过度表达进行无创定量成像。目标 2：使用小鼠中的人神经母细胞瘤异种移植物作为动物模型，评估使用 hCtrl 作为报告基因通过 Cu-64 PET 对 hCtrl 表达对肿瘤过表达 N-MYC 癌蛋白进行转录靶向成像的可行性。与其他报告基因相比，这种新的报告基因可能具有独特的理想特征：1）hCtrl不会刺激人类的免疫反应，预计对于非侵入性、定量地纵向监测转基因表达非常有用； 2) hCtrl报告基因预计对于前列腺癌基因治疗中靶向基因递送的成像特别有用，因为膀胱中几乎没有背景活动； 3) hCtrl 具有作为治疗基因的潜力，因为它能够调节细胞对顺铂的摄取。这项试点研究的数据将帮助我们确定使用 hCtrl 作为成像报告基因是否可行。这种新的 hCtrl 报告基因的成功开发有望为通过 PET 成像非侵入性和定量地纵向监测体内转基因表达提供多功能报告基因，从而显着促进人类基因治疗的进步。