Integration of Vascular Genomics and Proteomics for Diagnosis and Therapy of Canc

血管基因组学和蛋白质组学的整合在癌症诊断和治疗中的应用

• 批准号: 7905822
• 负责人: RENATA PASQUALINI
• 金额: $ 34.05万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-08 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7905822
• 关键词: Algorithms; Animal Model; Apoptosis; Automobile Driving; Bacteriophages; Bioinformatics; Biological; Biological Assay; Blood Vessels; Cells; Clinical; Companions; Data; Databases; Dependovirus; Development; Diagnosis; Diagnostic; Disease Progression; Effectiveness; Evaluation; Expressed Sequence Tags; Gene Expression; Gene Expression Profile; Genes; Genomics; Goals; Homing; Human Cell Line; Hybrids; Image; Imaging technology; In Vitro; Individual; Large-Scale Sequencing; Lead; Ligands; Malignant Neoplasms; Malignant neoplasm of prostate; Medicine; Modeling; Molecular Genetics; Monitor; Patients; Peptides; Pharmaceutical Preparations; Prostatic Neoplasms; Proteomics; Reporter; Reporter Genes; SAGE Library; Sampling; Suicide; System; Testing; Time; Transcript; Transgenes; Tumor Markers; United States Food and Drug Administration; Urine; Viral; Vision; base; cancer cell; cost; d(CH2)5(Tyr(Me)(2))AVP; design; genetic element; improved; in vivo; insight; molecular imaging; neoplastic cell; non-invasive monitor; particle; promoter; text searching; tool; tumor; tumor growth; vector

DESCRIPTION (provided by applicant): The integration of transcriptional profiling, cancer cells targeting and molecular-genetic imaging into a single platform would have many biological applications and the potential to improve the practice of medicine. We have designed and validated a new hybrid viral system composed of genetic elements from adeno-associated virus (AAV) and phage (termed AAVP). These ligand-directed particles enable targeted systemic delivery and imaging of transgene reporters. Experimental non-invasive monitoring of reporter trans-activation, may be followed ex-vivo and in vivo. Targeted molecular imaging would represent a major advance in the management of prostate cancer. The overall goal in this project is to combine the homing and tumor transducing capabilities of AAVP with cancer-specific promoters, as a way to develop improved tools for tumor monitoring based on the transcriptional activity of selected markers. These promoters will be identified by the large-scale evaluation of the transcriptome of cancer cells and tumor vasculature, using extensive searches in public databases, as well as by the construction and large-scale sequencing of SAGE libraries, using modern low-cost high throughput pyrosequencing approaches. The identified upregulated genes will be validated in two independent sample sets, and the promoters of confirmed upregulated transcripts will be identified, certified and cloned into AAVPs. With the definition of a reliable set of genes we expect to have comprehensive panel of tumor markers covering most of the gene expression variability seen in distinct patient tumor samples. The concept of establishing a panel of multiple markers is in line with the much heralded era of personalized medicine, and should launch, for the first time, a system for imaging temporally and spatially, in vivo, the transcriptional profile within tumors. The combination of a vector displaying peptides designed to target tumors, which also carries a suicide/reporter transgene (HSVtk) under the control of a tumor-specific promoter should enable transcriptional imaging and tumor growth suppression in a very specific fashion. The imaging and treatment possibilities of this vector reinforce the idea of drug-diagnostic co-development that, if preceded by an individual evaluation of gene expression (in the urine of patients with prostate cancer, for instance), could lead to a personalized diagnosis/treatment based on the up-regulated markers of an individual patient. This personalized imaging test merged with a companion drug comes together with the drug-diagnostic co-development concept recently put forward by the US Food and Drug Administration. Under this vision, the transcriptional imaging provided by a specific tumor gene will also trigger tumor apoptosis, allowing imaging as well as treatment and monitoring of disease progression. Our Specific Aims are: (i) To identify and to validate transcripts upregulated in prostate cancer using large scale transcriptome analysis. (ii) To combine ligand-directed targeting of AAVP to transcriptional targeting, and (iii) To evaluate the efficiency of imaging in vivo the expression of prostate cancer specific transcripts by using transcriptome-directed promoters cloned into RGD/GRP78-targeted AAVP constructs. The concept of establishing a panel of multiple markers is in line with the much heralded era of personalized medicine, and should launch, for the first time, a system for imaging temporally and spatially, in vivo, the transcriptional profile within tumors.

描述（由申请人提供）：将转录谱分析、癌细胞靶向和分子遗传成像整合到单一平台中将具有许多生物学应用和改善医学实践的潜力。我们设计并验证了一种新的杂交病毒系统，该系统由来自腺相关病毒（AAV）和噬菌体（称为AAVP）的遗传元件组成。这些配体导向的颗粒能够实现转基因报告基因的靶向全身递送和成像。报告子反式激活的实验性非侵入性监测可以在离体和体内进行。靶向分子成像将代表前列腺癌管理的重大进展。该项目的总体目标是将AAVP的归巢和肿瘤转导能力与癌症特异性启动子结合联合收割机，作为基于所选标记物的转录活性开发用于肿瘤监测的改进工具的一种方式。这些启动子将通过使用公共数据库中的广泛搜索对癌细胞和肿瘤脉管系统的转录组进行大规模评估，以及通过使用现代低成本高通量焦磷酸测序方法对SAGE文库进行构建和大规模测序来鉴定。鉴定的上调基因将在两个独立的样本集中进行验证，并且确认的上调转录本的启动子将被鉴定、认证并克隆到AAVP中。通过定义一组可靠的基因，我们期望有一组全面的肿瘤标志物，涵盖在不同患者肿瘤样本中观察到的大部分基因表达变异性。建立一组多个标记物的概念符合个性化医疗的时代，并且应该首次启动用于在时间和空间上在体内成像肿瘤内转录谱的系统。展示设计用于靶向肿瘤的肽的载体的组合，其还携带在肿瘤特异性启动子控制下的自杀/报告转基因（HSVtk），应该能够以非常特异的方式进行转录成像和肿瘤生长抑制。该载体的成像和治疗可能性强化了药物诊断共同开发的想法，如果在此之前对基因表达进行个体评估（例如，在前列腺癌患者的尿液中），可能会导致基于个体患者的上调标记物的个性化诊断/治疗。这种与伴随药物相结合的个性化成像测试与美国食品和药物管理局最近提出的药物诊断共同开发概念相结合。在这一愿景下，由特定肿瘤基因提供的转录成像也将触发肿瘤细胞凋亡，从而允许成像以及治疗和监测疾病进展。我们的具体目标是：（i）使用大规模转录组分析鉴定和验证前列腺癌中上调的转录本。(ii)结合联合收割机的配体导向靶向AAVP的转录靶向，和（iii）评估体内成像的效率，前列腺癌特异性转录本的表达，通过使用转录组定向启动子克隆到RGD/GRP 78靶向AAVP构建体。建立一组多个标记物的概念符合个性化医疗的时代，并且应该首次启动用于在时间和空间上在体内成像肿瘤内转录谱的系统。