The Biological and Clinical Relevance of EVI1 Expression in Prostate Carcinogenesis

EVI1 表达在前列腺癌发生中的生物学和临床相关性

• 批准号: 278607740
• 负责人: Professor Dr. Sven Perner
• 金额: --
• 依托单位: Institut für Pathologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/278607740?language=en
• 关键词: Biological; Clinical; Relevance; EVI1; Expression

Cancer stem cells (CSCs) are essential regulators of tumor initiation, progression and therapy resistance in cancer patients. Previous work from our and other groups indicate that the embryonic stem cell protein SOX2 is a CSC marker in ovarian, breast and prostate carcinoma (PCa), and furthermore plays oncogenic roles in lung and different types of squamous carcinomas. In this proposal, we propose to expand our knowledge on the molecular regulation of PCa stem cells by investigating the transcriptional stem cell regulator Ecotropic viral integration site 1 (EVI1). EVI1 has been mainly studied in healthy and malignant blood stem cells. In acute myeloid leukemia (AML), high EVI1 expression associates with particularly adverse clinical outcome. Even though EVI1 is expressed in several solid tumors including PCa, only few reports are available on EVI1 in solid tumors, and almost none in PCa. Preliminary data from our labs suggest that, in human PCa, EVI1 is a stem cell marker and enhances disease ag-gressiveness as well. We therefore propose to analyze expression, roles and molecular targets of EVI1 in PCa with particular focus on its relationship to PCa CSCs. We will analyze EVI1 protein expression in up to four independent PCa patient cohorts consisting either of primary tumors, localized lymph node and hormone-refractory distant metastases or only of primary tumors, and investigate whether high EVI1 ex-pression might represent a novel biomarker for PCa progression and prognosis. Next, we will explore whether amplification, translocation or activating mutations result in EVI1 protein overexpression. Using a lentiviral reporter system for the SOX2 regulatory regions 1 and 2 available in our lab, previously pub-lished phenotypic markers and aldehyde dehydrogenase (ALDH) activity assays, we will explore the as-sociation between EVI1 expression and established PCa stem cell markers. Potential functions mediated by EVI1 expression in PCa cells (including maintenance of PCa stem cell identity, growth, proliferation, apoptosis resistance, migration, invasion and in vivo tumorigenicity) will be explored using human PCa cells with modified EVI1 expression generated using lentiviral constructs. In vivo tumorigenicity assays will be performed in established NSG mouse xenograft assays, and respectively using a newly developed zebrafish xenograft model. Molecular target genes of EVI1 will be explored in PCa cells via RNA-Seq and ChIP analyses to identify drugable targets. Previously reported relevant target pathways (e.g. Smad3/TGF-beta, BCL proteins) and arsenic trioxide (ATO), which has recently been reported to reduce EVI1 protein stability in leukemic cells, will be explored for their efficacy to inhibit EVI1-mediated on-cogenic effects in PCa cells in vitro and eventually also in vivo.The data emerging from this project will improve our understanding of the molecular pathogenesis in PCa and contribute to the development of personalized treatment.

癌症干细胞（CSC）是癌症患者肿瘤发生、进展和治疗抗性的重要调节因子。我们和其他研究小组的先前工作表明，胚胎干细胞蛋白SOX 2是卵巢癌、乳腺癌和前列腺癌（PCa）中的CSC标志物，并且在肺癌和不同类型的鳞状细胞癌中发挥致癌作用。在这个建议中，我们建议扩大我们的知识PCa干细胞的分子调控研究转录干细胞调节嗜亲病毒整合位点1（EVI 1）。EVI 1主要在健康和恶性血液干细胞中进行研究。在急性髓性白血病（AML）中，高EVI 1表达与特别不良的临床结果相关。尽管EVI 1在包括PCa在内的几种实体瘤中表达，但关于EVI 1在实体瘤中的报道很少，在PCa中几乎没有。我们实验室的初步数据表明，在人类前列腺癌中，EVI 1是一种干细胞标志物，也能增强疾病的侵袭性。因此，我们建议分析EVI 1在PCa中的表达、作用和分子靶点，特别关注其与PCa CSC的关系。我们将分析EVI 1蛋白在多达四个独立的PCa患者队列中的表达，这些患者队列包括原发肿瘤、局部淋巴结和难治性远处转移或仅包括原发肿瘤，并研究EVI 1高表达是否可能代表PCa进展和预后的新生物标志物。接下来，我们将探讨扩增、易位或激活突变是否导致EVI 1蛋白过表达。利用我们实验室提供的SOX 2调控区1和2的慢病毒报告系统，先前发表的表型标记和醛脱氢酶（ALDH）活性测定，我们将探索EVI 1表达和已建立的PCa干细胞标记之间的关联。将使用具有使用慢病毒构建体产生的经修饰的EVI 1表达的人PCa细胞探索PCa细胞中EVI 1表达介导的潜在功能（包括维持PCa干细胞身份、生长、增殖、凋亡抗性、迁移、侵袭和体内致瘤性）。将在已建立的NSG小鼠异种移植试验中进行体内致瘤性试验，并分别使用新开发的斑马鱼异种移植模型进行体内致瘤性试验。将通过RNA-Seq和ChIP分析在PCa细胞中探索EVI 1的分子靶基因，以鉴定可药用靶标。先前报道的相关靶向通路（例如Smad 3/TGF-β，BCL蛋白）和三氧化二砷（ATO），最近报道其降低白血病细胞中的EVI 1蛋白稳定性，将探索其抑制EVI 1介导的对-本项目的数据将提高我们对PCa分子发病机制的理解，有助于个性化治疗的发展。

项目成果

Ecotropic viral integration site 1, a novel oncogene in prostate cancer

• DOI: 10.1038/onc.2016.325
• 发表时间: 2017-03
• 期刊: Oncogene
• 影响因子: 8
• 作者: Angela Queisser;S. Hagedorn;H. Wang;T. Schaefer;M. Konantz;S. Alavi;M. Deng;Wenzel Vogel;A. Mässenhausen;G. Kristiansen;S. Duensing;J. Kirfel;C. Lengerke;S. Perner