A novel molecular cross-talk driving pancreatic cancer progression

一种驱动胰腺癌进展的新型分子串扰

• 批准号: 10093980
• 负责人: Ajay Pratap Singh
• 金额: $ 34.66万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10093980
• 关键词: Apoptosis; Automobile Driving; Back; Binding; CCNE1 gene; CRISPR/Cas technology; Cell Hypoxia; Cell Line; Cell Proliferation; Cell Survival; ChIP-seq; Clinical; Data; Dependence; Development; Disease stratification; Exhibits; Face; Genes; Genetic Engineering; Genetic Transcription; Genomics; Growth; HIF1A gene; Histologic; Human; Hypoxia; Incidence; Knock-out; Knowledge; Lead; Life Expectancy; Luciferases; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Mediating; Mediator of activation protein; MicroRNAs; Molecular; Molecular Target; Neoplasm Metastasis; Null Lymphocytes; Pancreas; Pathogenesis; Pathway interactions; Patients; Phenotype; Play; Prevention; Preventive; Prognosis; Publishing; Quality of life; Regulation; Research; Resistance; Role; Sampling; Signal Pathway; Specificity; Stress; Stromal Neoplasm; Suggestion; System; TWIST1 gene; Testing; Therapeutic; Tissues; Up-Regulation; anticancer research; base; cancer diagnosis; clinically significant; improved; in silico; insight; mouse model; neoplastic cell; novel; novel strategies; nuclease; pancreatic cancer cells; pancreatic cancer patients; pancreatic neoplasm; promoter; response; transcriptional reprogramming; transcriptome; transcriptome sequencing; tumor; tumor hypoxia; tumor progression; v-myb Genes

Tumor cells inevitably face hypoxia during the course of their progression and their adaptation to hypoxic stress promotes invasive, metastatic and treatment-resistant phenotypes. Therefore, understanding the molecular basis underlying adaptive responses to hypoxia and identification of involved molecular targets will greatly facilitate the development of effective strategies for cancer management. We have recently provided first experimental evidence for a pathobiological role of MYB in pancreatic cancer (PC). Our novel preliminary findings now demonstrate i) role of MYB in hypoxic cell survival, ii), MYB-mediated regulation of HIF-1α, and iii) MYB-HIF-1α interaction and co-localization. In other novel findings, we show differential binding of MYB to its two target gene promoters under hypoxia. In addition, our novel data support the clinical significance of MYB by showing its wide-spread expression in pancreatic tumor cases, which is also suggestive of its association with increasing tumor-grade and patient's survival. Based on these compelling findings, we hypothesize that MYB-HIF1α crosstalk plays an important role in pancreatic cancer progression and metastasis, which will be tested in four specific aims. In aim 1, we will investigate the regulatory cross-talk between MYB and HIF-1α by studying coordinated regulation of MYB and HIF-1α under hypoxia, and any reciprocity that may exist between them. In aim 2, we will define the role of interaction between MYB and HIF-1α in their transcriptional reprogramming and hypoxia adaptive-response pathways. Specifically, we will examine if the MYB/HIF-1α crosstalk alters their genomic occupancy leading to changes in transcriptome, and characterize hypoxia adaptive-response phenotypes that are jointly or independently regulated by them. In aim 3, we will examine the cooperative functional significance of MYB and HIF-1α in pancreatic tumor progression and metastasis by using genetically-engineered, luciferase-tagged MYB- and HIF-1α expressing or knockout PC cells in an orthotopic mouse model. Histological and immunohistochemical studies will be performed to measure changes in tumor hypoxia, vasculature, cell proliferation/apoptosis, and metastasis. Finally, in aim 4, we will study the clinical significance of MYB-HIF-1α cross-talk in PC by performing immunohistochemical analysis in human pancreatic tumor samples along with adjacent and healthy normal pancreatic tissues to assess incidence, intensity and co-expression of MYB and HIF-1α. We will also examine their correlation (alone and in combination) with tumor -grade, -stage, and patient's survival. Together, these studies will deliver novel insight into the functional and mechanistic significance of a novel molecular cross-talk (MYB/HIF-1α) in PC pathobiology, and highlight its clinical significance. Resulting data would enhance our understanding of molecular pathogenesis of PC and, thus, facilitate the development of novel approaches for its prevention and treatment. Therefore, proposed studies have significant potential to impact pancreatic cancer research at various levels that will ultimately support effective management of this devastating malignancy.

肿瘤细胞在其发展和适应缺氧的过程中不可避免地面临缺氧 压力促进了侵袭性、转移性和治疗抗性表型。因此了解 低氧适应性反应的分子基础和相关分子靶点的鉴定将 大大促进了癌症管理的有效策略的发展。我们最近提供了 MYB在胰腺癌（PC）中的病理生物学作用的第一个实验证据。我们的小说初稿 现在的研究结果表明：i）MYB在缺氧细胞存活中的作用，ii）MYB介导的HIF-1α调节，iii） MYB-HIF-1α相互作用和共定位。在其他新的发现中，我们显示了MYB与其 两个目的基因启动子在缺氧条件下。此外，我们的新数据支持MYB的临床意义 通过显示其在胰腺肿瘤病例中的广泛表达，这也提示了其与胰腺癌的相关性。 随着肿瘤分级和患者存活率的增加。基于这些令人信服的发现，我们 假设MYB-HIF 1 α相互作用在胰腺癌进展和转移中起重要作用， 在四个具体目标中进行测试。在目的1中，我们将通过以下方法研究MYB和HIF-1α之间的调控串扰： 研究缺氧条件下MYB和HIF-1α的协调调节，以及MYB和HIF-1α之间可能存在的任何相互作用， 他们目的2：明确MYB和HIF-1α相互作用在MYB和HIF-1α转录调控中的作用。 重编程和缺氧适应性反应途径。具体来说，我们将检查MYB/HIF-1α是否 串扰改变了它们的基因组占有率，导致转录组的变化，并表征缺氧 适应性反应表型由它们共同或独立调节。在目标3中，我们将研究 MYB和HIF-1α在胰腺癌进展和转移中的协同作用 使用基因工程改造的、经腺苷三磷酸酶标记的MYB和HIF-1α表达或敲除的PC细胞， 原位小鼠模型。将进行组织学和免疫组织化学研究以测量变化 在肿瘤缺氧、脉管系统、细胞增殖/凋亡和转移中。最后，在目标4中，我们将研究 MYB-HIF-1α在PC中相互作用的临床意义 胰腺肿瘤样品沿着邻近和健康的正常胰腺组织以评估发病率， MYB和HIF-1α强度和共表达。我们还将检查它们的相关性（单独和在 组合）与肿瘤分级、分期和患者的存活率。总之，这些研究将提供新的见解， 探讨一种新的分子串扰（MYB/HIF-1α）在PC中的功能和机制意义 病理生物学，并强调其临床意义。由此产生的数据将增强我们对以下内容的理解 PC的分子发病机制，从而促进其预防和治疗的新方法的发展， 治疗因此，拟议的研究具有影响胰腺癌研究的重大潜力， 这将最终支持这种毁灭性恶性肿瘤的有效管理。