Transcription Factor Reporter Technology

描述(申请人提供)：对癌症基因表达异常调控的分子解剖揭示了癌症治疗的许多潜在靶点。这些靶点包括正常转录机制和异常转录机制的组成部分。由于许多信号转导途径在转录水平上的汇聚，参与癌症转录调控的蛋白质将获得高度优先考虑。针对转录靶点的新的分子疗法，包括siRNA技术，由于其干扰靶基因表达的精确度，比传统疗法具有显著的优势。虽然分子遗传学和药物化学的快速发展带来了新的衰减剂；但目前迫切需要开发能够对癌症对这些疗法的反应进行早期和非侵入性评估的技术。特别是，能够直接报告癌细胞中基因转录的成像技术对于癌症表型和分期以及评估新疗法都至关重要。近红外荧光的光学成像与酶特异性自猝灭探针的使用相结合，已成为活体癌细胞筛选的新技术。我们之前设计了一系列荧光探针，基于合成的生物相容的荧光染料载体，报告了荷瘤动物的水解性。最近，我们开发了新的化学方法来产生不对称和对称的寡核苷酸分子报告探针(ODMR)，旨在检测与转录因子核因子NFkappaB(NF-？B)的多效性和进化保守成分的相互作用。核因子-β通过调节肿瘤中细胞黏附、抗凋亡和细胞因子反应基因的表达，在肿瘤进展中发挥重要作用。我们最近测试了新的合成方法，将亲水性的、非干扰的连接物引入到B-box序列中，用于将荧光染料与这些探针和任何核苷磷酸共价结合。我们建议开发ODMR技术，这将是进一步推进癌症相关靶向转录激活剂体内成像的关键。

DESCRIPTION (provided by applicant): Molecular dissection of abnormal regulation of cancer gene expression has revealed many potential targets for cancer therapy. Those targets include the components of normal, as well as abnormal transcription machinery. Proteins involved in regulation of transcription in cancer will attain high priority due to the convergence of many signal transduction pathways at the transcriptional level. New molecular therapies directed to transcriptional targets, including siRNA technology have significant advantages over traditional therapies due to a precision of their interference with target gene expression. While rapid progress in molecular genetics and medicinal chemistry delivers new +attenuators; of gene expression, there is a critical need in developing technologies that enable early and non-invasive assessment of cancer response to these therapies. In particular, enabling imaging technologies that report directly on gene transcription in cancer cells are critically important for both cancer phenotyping and staging, as well as for evaluating new therapies. Optical imaging in the near-infrared range of fluorescence combined with the use of enzyme-specific self- quenched probes has emerged as novel technology of live cancer cell screening. We previously devised a family of fluorescent probes based on synthetic biocompatible carriers of fluorochromes that report on hydrolytic activity in tumor-bearing animals. Recently, we developed new chemistry for generating asymmetrical as well symmetrical oligonucleotide molecular reporter probes (ODMR) designed to sense interactions with pleiotropic and evolutionally conserved components of transcriptional factor nuclear factor NFkappaB (NF-?B). NF-?B plays one of the key roles in tumor progression by regulating expression of cell adhesion, antiapoptotic and cytokine responsive genes in cancer and stromal cells in tumors. We recently tested novel synthetic approaches for introducing hydrophilic, non-interfering linkers into ?B-box sequences for covalent binding of fluorochromes to these probes to any internucleoside phosphate. We propose to develop ODMR technology that will be essential for further advancement of in vivo imaging of cancer-related target transcription activators.