Alternative splicing and epithelial-mesenchymal plasticity in prostate tumors

前列腺肿瘤中的选择性剪接和上皮间质可塑性

• 批准号: 8196869
• 负责人: Mariano A. Garcia-Blanco
• 金额: $ 31.4万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8196869
• 关键词: Alternative Splicing; American Cancer Society; Behavior; Cells; Data; Disease; Distant; Epithelial; Epithelial Cells; Exons; Fibroblast Growth Factor Receptor 2; Gene Expression; Goals; Health; Human; Image; Malignant neoplasm of prostate; Mammals; Mesenchymal; Metastatic to; Molecular Profiling; Neoplasm Metastasis; New York; Outcome; Patients; Primary Neoplasm; Process; Property; Prostate; Prostate carcinoma; Prostatic Neoplasms; Protein Isoforms; RNA Splicing; Rattus; Regulation; Reporter; Reporting; Signal Transduction; System; Technology; Testing; Therapeutic; Time; Transcript; Tumor Markers; Vertebrates; Work; fitness; in vivo; killings; men; mouse model; neoplastic cell; novel; programs; therapeutic target; tumor; tumor progression; tumorigenesis

In epithelial cells alternative splicing of fibroblast growth factor receptor-2 (FGFR2) transcripts leads to the expression of the FGFR2(IIIb) isoform, whereas in mesenchymal cells the same process results in the synthesis of FGFR2(IIIc), which include exon IIIc. This regulated splicing is disrupted during progression of prostate carcinomas leading to the inappropriate expression of FGFR2 isoforms. To visualize the use of FGFR2 exon IIIc in Dunning AT3 tumors in syngeneic rats we constructed minigene constructs that report on alternative splicing. Imaging these alternative splicing decisions revealed unexpected mesenchymal-epithelial transitions (MET) in these primary tumors. The understanding of this epithelial-mesenchymal plasticity, revealed by differences in splicing, is the overarching goal of this application. To this end we propose to accomplish the following specific aims: Specific Aim 1. To characterize the connections between FGFR2 splicing and epithelial-mesenchymal plasticity in prostate tumors. We have developed a new bichromatic reporter that can be used to detect both MET and epithelial-mesenchymal transitions (EMT). We have also extended our observations to well characterized human prostate tumor cells in culture. Work accomplished in this aim will test the hypothesis that epithelial plasticity is a general property of aggressive prostate carcinomas in humans. Specific Aim 2. To characterize the epithelial-mesenchymal plasticity in primary prostate tumors revealed by alterations in splicing. In this aim we will characterize the molecular signatures and behavior of the MET clusters in primary tumors. Specific aim 3. To characterize the connections between epithelial-mesenchymal plasticity and metastatic behavior. We will investigate the connections between epithelial-mesenchymal plasticity and metastatic behavior. Finally, we will isolate circulating prostate tumor cells from both the AT3 rat tumors and patients, and we will determine their FGFR2 splicing status and their epithelial state. Work accomplished in aim two and in this aim will test the hypothesis that epithelial-mesenchymal plasticity leads to increased tumor fitness. The significance of this work is twofold: First we will unravel basic information about gene expression programs controlled by alternative splicing during prostate tumor progression. Second, the potential significance to human health is underscored by the magnitude of prostate cancer as a health problem. Identifying the tumors/cells that will progress to metastatic disease is a challenge and a priority. The work proposed here should identify useful markers and perhaps even therapeutic targets.

在上皮细胞中，成纤维细胞生长因子受体2（FGFR2）转录本的替代剪接 导致FGFR2（IIIB）同工型的表达，而在间充质细胞中相同的过程 导致FGFR2（IIIC）的合成，其中包括外显子IIIC。这种受调节的剪接被中断 在前列腺癌的进展过程中，导致FGFR2表达不当 同工型。为了可视化在合成大鼠中的Dunning AT3肿瘤中使用FGFR2外显子IIIC 构造的微型构造，以报告替代剪接。成像这些替代剪接 决策显示在这些原发性肿瘤中意外的间质上皮转变（MET）。 通过剪接差异揭示的对这种上皮 - 间质可塑性的理解是 此应用程序的总体目标。 为此，我们建议实现以下特定目标：特定目的1。 表征FGFR2剪接与上皮 - 间质塑性之间的连接 在前列腺肿瘤中。我们已经开发了一个新的双重记者，可用于检测两者 MET和上皮 - 间质转变（EMT）。我们还将观察结果扩展到了 在培养中表征了人类前列腺肿瘤细胞。在此目标中完成的工作将测试 假设上皮可塑性是侵略性前列腺癌的一般特性 人类。特定目的2。表征主要的上皮 - 间质可塑性 前列腺肿瘤通过剪接的改变揭示。在此目标中，我们将表征分子 原发性肿瘤中MET簇的特征和行为。特定目的3。 上皮 - 间质可塑性与转移行为之间的连接。我们将 研究上皮 - 间质塑性与转移行为之间的联系。 最后，我们将从AT3大鼠肿瘤和患者中分离出循环的前列腺肿瘤细胞，以及 我们将确定他们的FGFR2剪接状态及其上皮状态。目的完成的工作 第二，在此目标中将检验上皮 - 间质可塑性的假设增加 肿瘤健康。 这项工作的意义是双重的：首先，我们将揭开有关基因的基本信息 在前列腺肿瘤进展过程中由替代剪接控制的表达程序。第二， 前列腺癌的大小作为健康，对人类健康的潜在意义强调了 问题。识别将发展为转移性疾病的肿瘤/细胞是一个挑战， 优先事项。这里提出的工作应确定有用的标记，甚至可能是治疗性的 目标。