A Selective Approach to Metastatic Gene Discovery

描述（由申请人提供）：该提案旨在开发一种转移基因发现的选择性方法，并对PA-03-100有反应。在体外和体内直接选择在癌细胞中产生新的转移表型的转录因子的能力预计对我们理解转移的细胞生物学以及疾病的治疗具有显著影响。本文提出的研究利用了我们设计的能够激活或抑制内源性基因的转录因子的发展。现在可以制备以高亲和力和特异性识别9至18 bp DNA靶序列的多指锌指蛋白。当与激活或抑制结构域融合时，这些蛋白质成为靶基因转录活性的有效调节剂。目前没有其他基因疗法或基因工具策略提供既有效敲除又上调内源基因表达的手段。该提案的重点是使用我们的转录调节因子开发一种新的体内方法，用于在活体动物的背景下探索参与细胞归巢和转移的基因。一个全基因组的转录调控策略将被开发和应用于寻找参与转移和器官特异性归巢的基因。新的转移关键蛋白质的发现可能会导致一类新的癌症药物的开发，并有助于澄清土壤和种子转移假说。通过选择性和有效的转录调节因子，我们将解决设计的转录调节因子在动物模型中预防或以其他方式改变转移性疾病过程的潜力。这项研究的最终结果应该是增加对转移的细胞生物学的理解，并发现用于诊断、预后和治疗应用的新分子靶点。

DESCRIPTION (provided by applicant): This proposal seeks to develop a selective approach to metastatic gene discovery and is responsive to PA-03-100. The ability to directly select in vitro and in vivo for transcription factors that produce novel metastatic phenotypes in cancer cells is anticipated to have a significant impact on both our understanding of the cell biology of metastasis as well as the treatment of the disease. The study proposed here capitalizes on our development of designed transcription factors that enable activation or repression of endogenous genes. Polydactyl zinc finger proteins can now be prepared that recognize from 9 to 18 bp DMA target sequences with high affinity and specificity. When fused to activation or repression domains, these proteins become potent regulators of the transcriptional activity of the target genes. Currently no other gene therapy or gene tool strategy provides the means of both effectively knocking out and up-regulating the expression of an endogenous gene. This proposal focuses on the use of our transcriptional regulators to develop a new in vivo methodology for the exploration of genes involved in cell homing and metastasis in the context of the living animal. A genome-wide transcriptional modulation strategy will be developed and applied to the search for genes involved in metastasis and organ specific homing. The discovery of novel proteins key to metastasis could result in the development of a new class of cancer drugs and help to clarify the soil and seed metastasis hypothesis. With selective and potent transcriptional regulators we will address the potential of designed transcriptional regulators to prevent or otherwise alter the course of metastatic disease in animal models. The net result of this study should be an increased understanding of the cell biology of metastasis and the discovery of novel molecular targets for diagnostic, prognostic and therapeutic application.