In vivo reporters of gene expression

基因表达的体内报告基因

• 批准号: 8787876
• 负责人: Marit Nilsen-Hamilton
• 金额: $ 18.3万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8787876
• 关键词: Actins; Acute-Phase Proteins; Acute-Phase Reaction; Adverse effects; Aminoglycosides; Binding; Biopsy; Blood; Blood Proteins; Cells; Cellular biology; Chemicals; Detection; Deterioration; Failure; Gene Expression; Gene Expression Profile; Genes; Graft Rejection; Graft Survival; Health; Image; Immune; Immunosuppression; Immunosuppressive Agents; Incidence; Inflammation; Inflammatory; Inflammatory Response; Invaded; Kidney Transplantation; Life; Ligands; Location; Maintenance; Mammalian Cell; Measures; Molecular Biology; Monitor; Neomycin; Organ Transplantation; Organ failure; Organic Chemistry; Pharmaceutical Preparations; Probability; Procedures; Radiochemistry; Radiolabeled; Reporter; Reporter Genes; Research; Signal Transduction; Staging; Stem cells; Stimulus; Survival Rate; System; T-Lymphocyte; Testing; Tissues; Transplantation; Transplanted tissue; Variant; Veterans; aptamer; base; cellular imaging; in vivo; macrophage; novel strategies; prevent; promoter; public health relevance; radioligand; radiotracer; receptor; receptor binding; response; success; trafficking

DESCRIPTION (provided by applicant): Organ transplants are becoming increasingly common and the one year graft survival rates have risen sharply over the past 20 years due to aggressive immunosuppressive drug treatments. For example, renal transplants now have one year success rates of over 90% in most transplant centers. However, the long-term survival rate has not increased significantly over this same period. Subclinical inflammation (SCI) is a major contributor to transplant deterioration and to increase long-term success rates of organ transplants it is vital to have a measure of SCI in the transplant tissue. SCI is currently assesse by the invasive measure of a biopsy but could be more effectively evaluated by imaging of gene(s) that are activated during inflammation. We have developed a gene expression reporter system that can be used for imaging gene expression in deep tissue. The reporters are aptamers that are expressed from a chosen promoter and are visualized when they bind their target radioligand. The aptamers behave as intracellular receptors, binding their ligands tightly and thereby providing an image of the cells in which the promoter has been activated. The current veteran team, which includes expertise in cell and molecular biology, synthetic organic chemistry and radiochemistry, will develop an aptamer reporter system for imaging local inflammation of transplanted tissues by achieving the following specific aims: 1) Prepare mammalian cells that stably express neomycin aptamers as reporters of either a regulated (Lcn2) or constitutively expressed (actin) gene, 2) Develop the chemical procedures to synthesize and radiolabel aptamer ligands, and 3) Image actin and Lcn2 gene expression in vivo. The studies proposed here have broad implications for monitoring the health of tissue transplants to enable more informed decisions regarding medications. Such monitoring is anticipated to increase the survival of transplants that have been modified by the inclusion of the aptamer SCI reporter system prior to their transplantation. The same reporter platform may be applied to other imaging needs such as stem cell trafficking or their gene expression profile to monitor their differentiation stage.

描述（由申请人提供）：器官移植变得越来越普遍，由于积极的免疫抑制药物治疗，过去 20 年来移植物的一年存活率急剧上升。例如，现在大多数移植中心的肾移植一年成功率超过 90%。然而，同期长期生存率并未显着增加。亚临床炎症 (SCI) 是导致移植恶化的主要原因，为了提高器官移植的长期成功率，测量移植组织中的 SCI 至关重要。目前通过活检的侵入性测量来评估 SCI，但通过炎症期间激活的基因成像可以更有效地评估。我们开发了一种基因表达报告系统，可用于对深层组织中的基因表达进行成像。记者是从选定的启动子表达的适体，当它们结合目标放射性配体时被可视化。适体充当细胞内受体，紧密结合其配体，从而提供启动子已被激活的细胞的图像。目前的资深团队包括细胞和分子生物学、合成有机化学和放射化学方面的专业知识，他们将开发一种适体报告系统，用于对移植组织的局部炎症进行成像，通过实现以下具体目标：1）制备稳定表达新霉素适体作为调节（Lcn2）或组成型表达（肌动蛋白）基因报告的哺乳动物细胞， 2) 开发化学程序来合成和放射性标记适体配体，以及 3) 对体内肌动蛋白和 Lcn2 基因表达进行成像。这里提出的研究对于监测组织移植的健康状况具有广泛的意义，以便能够就药物做出更明智的决策。这种监测预计将提高移植物的存活率，这些移植物已通过纳入 移植前的适体 SCI 报告系统。相同的报告平台可应用于其他成像需求，例如干细胞运输或其基因表达谱以监测其分化阶段。