Targeting ASAP1-Controlled Signal Pathways to Inhibit Uveal Melanoma Metastasis

靶向 ASAP1 控制的信号通路抑制葡萄膜黑色素瘤转移

• 批准号: 10714245
• 负责人: Jae Hyuk Yoo
• 金额: $ 28.09万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-16 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714245
• 关键词: Ankyrin Repeat; Architecture; Binding; Biological Assay; Breast cancer metastasis; COUP transcription factor I; Cell Culture Techniques; Cell Nucleus; Cell membrane; Cells; Chromosome 3; Chromosomes; Country; Critical Pathways; Cytoplasmic Vesicles; Cytoskeleton; Data; Development; Disease; EMS1 gene; Exhibits; GNAQ gene; GTPase-Activating Proteins; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Growth; Human; In Vitro; Incidence; Invaded; Learning; Length; Liver; MAP Kinase Gene; Malignant Neoplasms; Mediating; Melanoma Cell; Metastatic Neoplasm to the Liver; Molecular; Molecular Target; Monomeric GTP-Binding Proteins; Monosomy; Mus; Mutation; Nebraska; Neoplasm Metastasis; Oncogenic; PH Domain; Pathway interactions; Patients; Persons; Process; Prognosis; Proliferating; Protein Kinase C; Proteins; Publishing; Research; Role; SH3 Domains; STAT4 gene; Signal Pathway; Signal Transduction; Testing; Transcription Factor AP-1; Tumor Promotion; Uveal Melanoma; Xenograft Model; Xenograft procedure; activating transcription factor 1; apoAI regulatory protein-1; beta catenin; cell motility; experimental study; human model; in vivo; inhibitor; insight; knock-down; malignant breast neoplasm; melanoma; mouse model; neoplastic cell; novel; novel strategies; overexpression; paxillin; pharmacologic; phospholipase C beta; pre-clinical; prevent; protein kinase C beta; trafficking; transcription factor; tumor; tumor progression; tumorigenesis; wound healing

Project Summary: "Targeting ASAP1-Controlled Signal Pathways to Inhibit Uveal Melanoma Metastasis" Uveal melanoma (UM) accounts for about 5% of all melanomas and has an incidence of 4-11 per million people per year in western countries. Although much has been learned about the signaling pathways that control UM oncogenesis, far less is known about the pathways that control metastatic disease. Preventing or effectively treating metastatic disease is critical because it occurs in about 50% of patients (most often in the liver) and once present, the prognosis is dismal. Amplification of chromosome 8q is one of the strong predictors of metastatic disease. ASAP1, which is located on chromosome 8q, has been shown to control metastasis in breast cancer. ASAP1 is amplified in about 40% of UM patients, and increased ASAP1 expression levels are associated with increased metastasis and poor prognosis. Overexpression of ASAP1 in low-grade tumor cells increases cell migration in an in vitro wound-healing assay. Furthermore, ASAP1 has been implicated in the metastasis of several cancers and is thought to promote metastasis by directly controlling the remodeling of the cytoskeletal architecture. We have recently begun to explore the molecular pathways that control uveal melanoma metastasis and have preliminary data showing that ASAP1 controls uveal melanoma cell invasion, proliferation, and metastasis in an orthotopic xenograft model of human uveal melanoma. Furthermore, we have identified several tumor-associated transcription factors, including STAT4, COUP-TF I, and COUP-TF II, that are activated by overexpression of ASAP1 in UM cells, suggesting a novel mechanism by which ASAP1 may also control UM metastasis. Therefore, we hypothesize that ASAP1 promotes uveal melanoma metastasis by activating signaling pathways that induce tumor-associated transcription factors and that inhibiting these pathways by pharmacologic inhibition will significantly reduce metastatic disease. We will test this hypothesis by pursuing two aims. In Aim 1, we will identify ASAP1 domain(s) that promote UM cell invasion and proliferation. These studies will involve expressing full-length ASAP1 and its serial deletion constructs following ASAP1 knockdown to determine the role of ASAP1 domains in UM cells. In Aim 2, we will assess the function of ASAP1-controlled transcription factor(s) in uveal melanoma cell invasion, proliferation, and tumor metastasis. Cell culture experiments will involve either knockdown of gene expression or pharmacological inhibition to determine the role of these transcription factors in UM cells. In vivo experiments will determine whether pharmacologic inhibition of ASAP1- activated transcription factors reduce tumor progression and metastasis in a xenograft model of human UM. For these studies, we will use JAK inhibitors and CIA1, which inhibit STAT4 and COUP-TF II, respectively. This research should provide us with new insights into the molecular mechanisms underlying metastatic spread of UM and may lead to novel approaches to prevent or treat metastatic UM.

项目摘要：“针对ASAP1控制的信号途径靶向抑制紫veal黑色素瘤 转移” 卵子黑色素瘤（UM）约占所有黑色素瘤的5％，发病率为4-11 西方国家每年的人。尽管已经了解了很多有关信号通路 控制UM肿瘤发生，对控制转移性疾病的途径知之甚少。预防或 有效治疗转移性疾病至关重要，因为它发生在约50％的患者中（最常见于 肝脏），一旦存在，预后就令人沮丧。 8q染色体的扩增是强的 转移性疾病的预测因子。位于8Q染色体上的ASAP1已显示可以控制 乳腺癌的转移。 ASAP1在大约40％的UM患者中被扩增，并增加了ASAP1 表达水平与转移和预后不良有关。 ASAP1的过表达 在低级肿瘤细胞中，在体外伤口治疗测定中增加了细胞迁移。此外，ASAP1 已经与几种癌症的转移有关，被认为可以直接促进转移 控制细胞骨架架构的重塑。我们最近开始探索分子 控制卵巢黑色素瘤转移并具有初步数据的途径，表明ASAP1控制 紫veal黑色素瘤细胞侵袭，增殖和转移，在人卵子的原位异种移植模型中 黑色素瘤。此外，我们已经确定了几个肿瘤相关的转录因子，包括STAT4， 由UM细胞中ASAP1的过表达激活的COUP-TF I和COUP-TF II，这表明是一种新颖的 ASAP1也可以控制UM转移的机制。因此，我们假设ASAP1 通过激活诱导肿瘤相关的信号传导途径来促进紫veal黑色素瘤转移 转录因子以及通过药理抑制抑制这些途径的因素将显着降低 转移性疾病。我们将通过实现两个目标来检验这一假设。在AIM 1中，我们将确定ASAP1 促进UM细胞侵袭和增殖的结构域。这些研究将涉及表达全长 ASAP1及其串行删除构建体后ASAP1敲低以确定ASAP1的作用 UM细胞中的域。在AIM 2中，我们将评估ASAP1控制转录因子的功能 卵巢黑色素瘤细胞侵袭，增殖和肿瘤转移。细胞培养实验将涉及 敲低基因表达或药理学抑制以确定这些转录的作用 UM细胞中的因素。体内实验将确定药物学抑制ASAP1是否激活 转录因子在人类UM的异种移植模型中降低了肿瘤的进展和转移。为此 研究，我们将使用JAK抑制剂和CIA1分别抑制STAT4和COUP-TF II。这 研究应为我们提供对分子机制的新见解 UM并可能导致预防或治疗转移性UM的新方法。