A METHOD TO IDENTIFY UPSTREAM REGULATORS OF ONCOGENES

描述（由申请人提供）： 尽管付出了巨大的努力，但对每种原发性肿瘤的所有转导伙伴、转录因子及其靶点缺乏全面的了解。此外，只存在有限数量的已知的“监管趋同点”，涵盖多种癌症类型。Vaccinex开发了一种高效的专有技术，用于在哺乳动物背景下基于功能选择识别基因。该方法利用插入到牛痘病毒（vv）基因组中的cDNA文库，并已成功地应用于分离新的肿瘤抗原。在牛痘病毒中制备文库的能力代表了一个主要的进步，因为牛痘病毒是唯一的哺乳动物表达载体，其中可以构建多样的和代表性的cDNA文库，其元件可以有效地从感染细胞的细胞质中回收。我们建议应用这种功能性基因选择技术的变体，使用已知的癌基因和肿瘤细胞标记物作为“诱饵”来识别调节其活性的蛋白质。 这种方法代表了一个令人耳目一新的，功能的替代全球转录监测实验进行cDNA和寡核苷酸微阵列。具体而言，微阵列实验监测与细胞环境相关的全局转录事件，我们的技术选择了一个明确的转化终点，无论是以上调的转录本或过度丰富的蛋白质的形式，并提出揭示调节这些目标的蛋白质。因此，Vaccinex技术具有在转导途径中功能性扫描上游的独特能力。此外，该技术旨在从重叠的途径中识别共同的调节点。直接在癌症背景下定义这些调节机制是产生更有效治疗方法的关键步骤。

DESCRIPTION (provided by applicant): A comprehensive understanding of all transduction partners, transcription factors, and their targets responsible for each type of primary tumor is lacking, despite intense effort. Furthermore, only a limited number of known "points of regulatory convergence," covering multiple cancer types, exists. Vaccinex has developed a highly efficient, proprietary technology for the identification of genes based on functional selection within a mammalian background. This method utilizes cDNA libraries inserted into the vaccinia virus (vv) genome and has been successfully applied to the isolation of novel tumor antigens. The ability to make libraries in vaccinia virus represents a major advance, since vaccinia virus is the only mammalian expression vector in which it is possible to construct a diverse and representative cDNA library whose elements can be efficiently recovered from the cytoplasm of infected cells. We propose to apply a variant of this functional gene selection technology to use known oncogenes and tumor cell markers as 'bait' to identify proteins that modulate their activity. This approach represents a refreshing, functional alternative to global transcript monitoring experiments performed with cDNA and oligonucleotide microarrays. Specifically, where microarray experiments monitor global transcriptional events related to cellular environment, our technique chooses a well-defined end point of transformation, either in the form of an up-regulated transcript or an over-abundant protein and proposes to uncover proteins that regulate these targets. Vaccinex technology, therefore, has the unique capability to functionally scan upstream in a transduction pathway. Moreover, the technique is designed to identify common points of regulation from overlapping pathways. Defining these regulatory mechanisms directly in the context of cancer is a critical step for generating more effective therapeutics.