Investigating GLI transcription factors as regulators of the pancreatic cancer microenvironment

研究 GLI 转录因子作为胰腺癌微环境的调节剂

• 批准号: 10681714
• 负责人: Benjamin Allen
• 金额: $ 59.12万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10681714
• 关键词: Advanced Development; Alleles; Biological Assay; Cancer Etiology; Cell Line; Cells; Cessation of life; Chromatin; Clinical Trials; Cre driver; Data; Diagnosis; Disease; Disease Progression; Epitopes; Erinaceidae; Family; Fibroblasts; GLI gene; Genetic; Genetic Models; Genetic Transcription; Glioma; Growth; Health; Human; Immune; In Vitro; Incidence; Individual; Knock-in; Laboratories; Ligands; Macrophage; Malignant Neoplasms; Malignant neoplasm of pancreas; Migration Assay; Mus; Neoplasm Metastasis; Organoids; Pancreas; Pathway interactions; Primary Neoplasm; Proteins; RNA; Regulation; Role; Signal Transduction; Signaling Protein; Speed; Survival Rate; T cell infiltration; T-Lymphocyte; Testing; Transcriptional Regulation; Transplantation; Tumor-Derived; United States; Work; cancer therapy; cell motility; effective therapy; genetic approach; improved; in vivo; inhibitor; mouse model; novel; pancreatic cancer model; pancreatic neoplasm; paracrine; patient prognosis; pharmacologic; smoothened signaling pathway; transcription factor; transplant model; tumor; tumor growth; tumor microenvironment; tumor progression

PROJECT SUMMARY/ABSTRACT Pancreatic cancer is a deadly human malignancy, with a five-year survival rate of 11%. Further, the incidence of pancreatic cancer is rising and is projected to be the second-leading cause of cancer-related death in the United States by 2030. The increasing occurrence of this disease, combined with the lack of effective treatments, creates an urgent need to better understand the factors that contribute to the progression of pancreatic cancer, and to develop improved therapies that will tangibly benefit human health. One pathway that has been implicated in pancreatic cancer for nearly twenty years is the Hedgehog (HH) signaling pathway. Initial work demonstrated that HH ligand expression increases during pancreatic cancer progression, while more recent studies have found that tumor-derived HH ligands signal in a paracrine fashion to activate HH signaling in pancreatic fibroblasts. As with many efforts to treat pancreatic cancer, clinical trials with HH pathway inhibitors have proven ineffective, or actually sped disease progression. However, work from our laboratories has identified three novel findings regarding the role of HH signaling in pancreatic cancer progression: First, we have found that the levels of HH signaling can determine whether HH signaling promotes or inhibits pancreatic tumor growth; Second, we have recently discovered that HH pathway inhibition can alter the fibroblast composition in the pancreatic cancer microenvironment; Third, our data indicate that HH pathway inhibition dramatically alters immune composition, through altered fibroblast-immune crosstalk. Together, these data suggest that an improved understanding of the role of HH signaling in the pancreatic cancer microenvironment, including HH-dependent fibroblast-immune crosstalk will provide an opportunity to advance the development of more effective pancreatic cancer treatment options. In this proposal, we will investigate the role of glioma-associated proteins (GLIs), a family of three proteins (GLI1-3) that are the transcriptional effectors of the HH signaling pathway, in pancreatic cancer progression. Specifically, we will use novel pancreatic fibroblast cell lines lacking Gli1, Gli2 or Gli3 individually and in combination to investigate the consequences on pancreatic tumor growth in co-transplantation assays. We will also investigate Gli deletion in vivo using two different Cre driver alleles (PdgfraCreER and Gli1CreER) in a novel, FlpO-driven mouse model of pancreatic cancer progression. Further, we will investigate GLI- dependent effects on fibroblast-immune crosstalk through assessment of macrophage and T cell migration, polarization and function. Finally, we will define GLI transcriptional targets in pancreatic fibroblasts both in vitro and in vivo, in the normal pancreas and in the context of pancreatic cancer, using a novel set of epitope-tagged knock-in alleles. This work will advance our understanding of the role of HH/GLI signaling in pancreatic cancer.

项目摘要/摘要 胰腺癌是一种致命的人类恶性肿瘤，五年生存率为11％。此外，发病率 胰腺癌正在上升，预计将成为联合癌症相关死亡的第二个领先原因 到2030年。 迫切需要更好地了解胰腺癌进展的因素， 并开发改进的疗法，将有明显地使人类健康受益。一条牵连的途径 在胰腺癌中，近二十年是刺猬（HH）信号通路。最初的工作证明了 在胰腺癌进展过程中，HH配体的表达增加，而最近的研究发现 肿瘤衍生的HH配体以旁分泌方式信号，以激活胰腺成纤维细胞中的HH信号传导。作为 在许多治疗胰腺癌的努力下，用HH途径抑制剂进行的临床试验已被证明无效或 实际上是促进疾病的进展。但是，我们实验室的工作已经确定了三个新颖的发现 关于HH信号在胰腺癌进展中的作用：首先，我们发现HH的水平 信号传导可以确定HH信号传导是否促进或抑制胰腺肿瘤的生长；第二，我们有 最近发现，HH途径抑制可以改变胰腺癌的成纤维细胞组成 微环境；第三，我们的数据表明HH途径抑制作用显着改变了免疫组成， 通过改变的成纤维细胞免疫串扰。这些数据一起表明，对 HH信号在胰腺癌微环境中的作用，包括HH依赖性成纤维细胞免疫 Crosstalk将提供一个机会，以促进开发更有效的胰腺癌治疗 选项。在此提案中，我们将研究与神经胶质瘤相关蛋白（GLI）的作用，这是一个三口之家 蛋白质（GLI1-3）是胰腺癌中HH信号通路的转录效应子 进展。具体而言，我们将使用缺乏GLI1，GLI2或GLI3的新型胰成纤维细胞系 并结合研究共转移测定中胰腺肿瘤生长的后果。 我们还将使用两个不同的CRE驱动等位基因（PDGFRACREER和GLI1CREER）在体内研究GLI缺失 在一种新型的胰腺癌进展的FLPO驱动小鼠模型中。此外，我们将研究光明 通过评估巨噬细胞和T细胞迁移，对成纤维细胞 - 免疫串扰的影响， 极化和功能。最后，我们将在体外定义胰腺成纤维细胞中的GLI转录靶标 在体内，在正常的胰腺和胰腺癌的背景下，使用一组新型表位标签 敲入等位基因。这项工作将促进我们对HH/GLI信号在胰腺癌中的作用的理解。