IDENTIFYING GERMLINE GENES AS TARGETS FOR THERAPEUTIC INTERVENTION IN CANCER

确定种系基因作为癌症治疗干预的目标

• 批准号: 8554776
• 负责人: MARY LOU KING
• 金额: $ 14.52万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-27 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8554776
• 关键词: Antineoplastic Agents; Apoptosis; Area; Balbiani Body; Behavior; Biochemical; Biological; Biological Assay; Brain; Cadherins; Cancer Biology; Cancer cell line; Categories; Cell Cycle; Cell Death; Cell Lineage; Cell division; Cell-Cell Adhesion; Cellular Structures; Characteristics; Data; Development; Drosophila genus; Embryo; Epithelial; Gene Expression; Genes; Genetic; Germ; Germ Cell Cancers; Germ Cells; Goals; Immunity; Liquid Chromatography; Malignant Neoplasms; Mesenchymal; Mitochondria; Neoplasm Metastasis; Oocytes; Oogenesis; Organism; Outcome; Process; Proteins; RNA; RNA Sequence Analysis; Research; Seminal; Source; Specific qualifier value; Structure; Technology; Testing; Therapeutic Intervention; To specify; Tumor Suppressor Genes; Vertebrates; Work; Xenopus; anticancer research; base; cancer stem cell; cancer therapy; drug development; expression cloning; gene function; neuronal cell body; novel; novel marker; public health relevance; scaffold; screening; stem cell biology; tandem mass spectrometry; therapeutic target; tumor; tumor growth

DESCRIPTION (provided by applicant): Recent progress in cancer research has highlighted a new category of targets for anti-cancer drugs: gene products expressed in the germline (Janic et al., 2010; Georlette et al., 2007; Wu and Ruvkun, 2010; Liu et al., 2011; Strumane et al., 2006; Caballero et al., 2009). Seminal work in Drosophila has shown that loss of a tumor suppressor gene causes brain malignancies that display a soma-to-germline transformation with 25% of up-regulated genes having a germline-associated function (Janic et al., 2010). Many of these tumor up-regulated genes are of maternal origin. Importantly, blocking the expression of these genes suppressed tumor growth. Germ cells share at least three important characteristics of cancer, the ability to: sustain a proliferative state, resist cell death, and undergo an epithelial-mesenchymal transition characteristic of invasion and metastasis. These findings strongly support the value of screening germ cell components for their ability to promote these behaviors as a step towards identifying potent therapeutic targets for cancer treatments. However, previous work on identifying germline components has relied almost entirely on RNA microarray data while the proteins remain largely unknown and the majority of RNAs uncharacterized, slowing progress in this area (Yatsu et al., 2008; Molyneaux et al., 2004; Ewen and Koopman, 2010). Germ plasm is the subcellular domain unique to germ cell precursors that contains all the determinants required to specify the germ cell lineage in diverse organisms. Our central hypothesis is that germ plasm will provide a rich source of new targets for anti-cancer drugs. Unfortunately, it is technically difficult to isolate germ plasm in sufficient quantities for proten analysis and thus, genetic information in vertebrates has been limited. Xenopus offers a unique opportunity to isolate biochemical quantities of germ plasm, making it possible, with current technology, to create a complete "parts list" of this cellular "machine" that specifies the totipotnt germ cell lineage. Moreover, Xenopus is highly amenable to expression cloning, allowing a high-throughput approach to assess gene function of germ plasm components. To test our central hypothesis, we will complete the following two specific aims: Aim 1. Identify the protein and RNA components of germ plasm and use this information to predict gene networks operating in the germline and up-regulated in cancer cell lines. Aim 2. Test these germ cell components for their ability to promote the biological "hallmarks" of cancer: metastasis, immortality, and proliferation in bioassays. In preliminary studies, we have isolated biochemical amounts of germ plasm and have identified over 400 proteins by Liquid Chromatography-tandem Mass Spectrometry analyses. We are now ready to launch into broader studies functionally screening germ plasm components for their possible relevance to cancer biology.

描述（由申请人提供）：近年来癌症研究的进展突出了一类新的抗癌药物靶点：生殖系表达的基因产物（Janic等，2010；Georlette等，2007；Wu和Ruvkun, 2010； Liu等，2011；Strumane等，2006；Caballero等，2009）。对果蝇的开创性研究表明，肿瘤抑制基因的缺失导致脑恶性肿瘤表现为体细胞向种系转化，其中25%的上调基因具有种系相关功能（Janic等，2010）。这些肿瘤上调基因中有许多来自母亲。重要的是，阻断这些基因的表达可以抑制肿瘤的生长。生殖细胞至少具有癌症的三个重要特征：维持增殖状态，抵抗细胞死亡，经历侵袭和转移的上皮-间质转化特征。这些发现有力地支持了筛选生殖细胞成分的价值，因为它们能够促进这些行为，这是确定癌症治疗的有效治疗靶点的一步。然而，先前鉴定生殖系成分的工作几乎完全依赖于RNA微阵列数据，而蛋白质在很大程度上仍然未知，大多数RNA未被表征，这减缓了该领域的进展（Yatsu等人，2008；Molyneaux等人，2004；Ewen和Koopman， 2010）。种质是生殖细胞前体特有的亚细胞结构域，它包含了在不同生物体中指定生殖细胞谱系所需的所有决定因素。我们的中心假设是，种质将为抗癌药物的新靶点提供丰富的来源。不幸的是，技术上很难分离出足够数量的种质进行蛋白质分析，因此，脊椎动物的遗传信息受到限制。Xenopus提供了一个独特的机会来分离生物化学数量的种质，使之成为可能，用目前的技术，创建一个完整的“部件列表”的细胞“机器”，指定全能的生殖细胞谱系。此外，非洲爪蟾具有高度的表达克隆性，这使得高通量方法可以评估种质成分的基因功能。为了验证我们的中心假设，我们将完成以下两个具体目标：目标1。鉴定种质的蛋白质和RNA成分，并利用这些信息来预测在种系中运行和在癌细胞系中上调的基因网络。目标2。在生物检测中测试这些生殖细胞成分促进癌症生物学“特征”的能力：转移、不朽和增殖。在初步研究中，我们已经分离了种质的生化量，并通过液相色谱-串联质谱分析鉴定了400多种蛋白质。我们现在准备开展更广泛的研究，从功能上筛选种质成分，以寻找它们与癌症生物学的可能相关性。