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: 7579408
• 负责人: Mariano A. Garcia-Blanco
• 金额: $ 32.37万
• 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/7579408
• 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; Left; 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; public health relevance; therapeutic target; tumor; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The potential significance of our application to human health is underscored by the magnitude of prostate cancer as a health problem, which The American Cancer Society (ACS) predicts will afflict ~200,000 men and will kill >25,000 in the US alone. In addition, prostate cancer presents the clinician and the patient with unique problems in terms of choosing among therapeutic options and predicting eventual outcomes (see New York Times 26 February article "A Review of Prostate Cancer Leaves Men in a Muddle"). The work proposed here should identify useful markers of tumor behavior and perhaps even targets for novel therapies.

描述（由申请人提供）：在上皮细胞中，成纤维细胞生长因子受体-2（FGFR2）转录物的替代剪接导致FGFR2（IIIB）同工型的表达，而在间质细胞中，相同的过程导致FGFR2（IIIC）的合成，其中包括Exon IIIC。 在前列腺癌的进展过程中，这种调节的剪接会破坏，从而导致FGFR2同工型的不适当表达。 为了可视化在合成大鼠中的Dunning AT3肿瘤中使用FGFR2外显子IIIC，我们构建了有关替代剪接的微型结构。 成像这些替代剪接决策显示出意外的间质上皮跃迁（MET）在这些原发性肿瘤中。 通过剪接差异揭示的对这种上皮 - 间质可塑性的理解是该应用的总体目标。 为此，我们建议实现以下特定目的：特定目的1。表征前列腺肿瘤中FGFR2剪接与上皮 - 间质可塑性之间的连接。 我们已经开发了一个新的双分型报告基因，该报告剂可用于检测MET和上皮 - 间质转变（EMT）。 我们还将观察结果扩展到了培养中人类前列腺肿瘤细胞的良好表征。 在此目标中完成的工作将检验以下假设：上皮可塑性是人类侵略性前列腺癌的一般特性。 特定目的2。表征原发性前列腺肿瘤中的上皮 - 间质可塑性，揭示了剪接的改变。 在此目的中，我们将表征原发性肿瘤中MET簇的分子特征和行为。 特定目的3。表征上皮 - 间质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循环前列腺肿瘤细胞，并确定其FGFR2剪接状态及其上皮状态。 在目标二中完成的工作将测试上皮 - 间质可塑性导致肿瘤适应性增加的假设。 这项工作的意义是双重的：首先，我们将在前列腺肿瘤进展过程中通过替代剪接控制的基因表达程序进行基本信息。 其次，对人类健康的潜在意义被前列腺癌的大小作为健康问题所强调。 识别将发展为转移性疾病的肿瘤/细胞是一个挑战和优先级。 这里提出的工作应确定有用的标记，甚至应该确定治疗目标。 公共卫生相关性：我们对人类健康的应用的潜在意义强调了前列腺癌作为一种健康问题的大小，美国癌症学会（ACS）预测，仅在美国，仅在美国就将杀死约25,000人。 此外，前列腺癌为临床医生和患者提供了在治疗选择中选择和预测最终结果的独特问题（请参阅2月26日的2月26日的文章“对前列腺癌的评论使男性陷入混乱”）。 这里提出的工作应确定肿瘤行为的有用标记，甚至可以确定新疗法的目标。