Role of Cancer-Associated Fibroblasts in Cholangiocarcinoma

癌症相关成纤维细胞在胆管癌中的作用

基本信息

批准号：
10166796
负责人：
Xin Chen
金额：
$ 62.16万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-19 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10166796
关键词：
Ablation Affect Apoptotic Blocking Antibodies Cell Compartmentation Cell Death Cell Proliferation Cells Characteristics Cholangiocarcinoma Clinical Clinical Research Coculture Techniques Collagen Collagen Receptors Desmoplastic Development Drug usage Extracellular Matrix FDA approved Family Fibroblasts Fibrosis Gene Expression Genes Goals Growth Human In Vitro Incidence Injections Integrins Interruption Intrahepatic Cholangiocarcinoma Knock-out Knockout Mice Light Link Liver Liver neoplasms Malignant Neoplasms Malignant neoplasm of pancreas Measures Mediating Mediator of activation protein Mus Myofibroblast Nature Outcome Pancreas Pathway interactions Pharmaceutical Preparations Pharmacology Platelet-Derived Growth Factor Primary Malignant Neoplasm of Liver Protein Isoforms Role Sampling Signal Pathway Signal Transduction Survival Rate System Testing Therapeutic Time Tumor Promoters Tumor Promotion Tumor-Derived cancer genetics conventional therapy discoidin domain receptor 1 discoidin receptor experimental study genetic approach in vivo inhibiting antibody innovation interest intrahepatic liver injury migration neoplastic cell new therapeutic target novel novel therapeutic intervention recruit single cell sequencing single-cell RNA sequencing tool tumor tumor microenvironment

项目摘要

Intrahepatic cholangiocarcinoma (ICC) is an aggressive liver tumor with limited therapeutic options and 5-year survival rates of less than 10%. ICC is characterized by its highly desmoplastic nature, with abundance of cancer- associated fibroblasts (CAF) and extracellular matrix (ECM). The role of CAF and ECM in ICC remain controversial due to the paucity of functional in vivo studies. While the majority of ICC in vitro studies support a cancer-promoting role of CAF, recent studies in endogenously arising pancreatic cancer, a highly desmoplastic tumor with many similarities to ICC, have shown that CAF restrain cancer growth. Here, we seek to answer the question whether CAF promote or restrain ICC, using a number of novel and cell-specific tools to manipulate CAF and tumor cells and their crosstalk in endogenously arising ICC in vivo. We hypothesize that CAF and ECM provide a niche that promotes ICC growth and survival, and that detailed characterization of ICC-CAF crosstalk will identify novel therapeutic targets within the tumor microenvironment. In Aim 1, we will determine the role CAF using novel tools to determine how genetic CAF inhibition or early and late CAF ablation affect ICC growth, proliferative and anti-apoptotic signaling pathways, and mouse survival. In Aim 2, we will investigate pathways that mediate the recruitment, proliferation and activation of CAF in ICC focusing on the hypothesis that tumor cells hijack normal fibrogenic mechanism in the liver via tumor-derived TGFb and PDGF isoforms and TGFb- activating integrins, resulting in accumulation and activation of CAF. In addition to detailed mechanistic studies in knockout mice and in vitro co-cultures, we will determine whether pharmacologic inhibition of CAF activation by FDA-approved drug Nintedanib or integrin-blocking antibodies inhibit ICC growth and prolong mouse survival. In Aim 3, we will determine pathways through which CAF modulate ICC growth, focusing on the hypothesis that CAF-derived ECM activates tumor-promoting signals in the tumor cell compartment. To test this hypothesis, we will investigate ICC development in mice with CAF-specific knockout of Col1a1, or tumor-selective knockout of collagen-sensing receptor discoidin domain receptor 1 (DDR1). In addition, we will determine whether ECM- mediated stiffness and subsequent activation of mechanosensitive signaling in tumor cells promote ICC development. We will measure tumor stiffness, activation of mechanosensitive signaling pathways and DDR1 expression in human CCA samples and correlate these to clinical outcomes. The role of stiffness and DDR1 in ICC growth and CAF-ICC crosstalk will be investigated in more detail in vitro through modulation of stiffness and by co-culturing CAF and ICC cells that lack Col1a1 or DDR1, respectively. In summary, the proposed studies will employ novel tools to answer a long-standing question in the field and may provide a basis for targeting ICC- CAF crosstalk as novel therapeutic strategy for this devastating malignancy.

肝内胆管癌（ICC）是一种侵略性的肝肿瘤，治疗选择有限和5年存活率小于10％。 ICC的特征是其高度塑性性质，并以癌症抽象为特征。相关的成纤维细胞（CAF）和细胞外基质（ECM）。 CAF和ECM在ICC中的作用仍然存在由于体内研究的功能缺乏，引起了争议。虽然大多数ICC体外研究支持 CAF的癌症促进作用，最近在内源性胰腺癌中进行的研究，一种高脱塑料与ICC相似的肿瘤表明CAF抑制了癌症的生长。在这里，我们看到要回答疑问CAF是否使用许多新颖和细胞特异性工具来促进或限制ICC进行操纵 CAF和肿瘤细胞以及内源性的ICC体内产生的crostalk。我们假设CAF和ECM 提供一个促进ICC增长和生存的利基市场，以及ICC-CAF Crosstalk的详细表征将确定肿瘤微环境中的新型治疗靶标。在AIM 1中，我们将确定角色 CAF使用新颖的工具来确定遗传CAF抑制作用或早期和较晚的CAF消融如何影响ICC的增长，增殖和抗凋亡信号通路以及小鼠存活。在AIM 2中，我们将研究途径介导ICC中CAF的募集，增殖和激活，重点是肿瘤的假设细胞通过肿瘤衍生的TGFB和PDGF同工型和TGFB-劫持肝脏中的正常纤维化机制激活整联蛋白，导致CAF的积累和激活。除了详细的机械研究在基因敲除小鼠和体外共培养中，我们将确定CAF激活的药理抑制通过FDA批准的药物nintedanib或整联蛋白阻断抗体抑制了ICC生长和延长小鼠的存活。在AIM 3中，我们将确定CAF调节ICC增长的途径，重点是以下假设。 CAF来源的ECM激活肿瘤细胞室中的肿瘤信号。为了检验这一假设，我们将研究CAF特异性敲除COL1A1或肿瘤选择性敲除小鼠的ICC发育胶原蛋白感应受体盘状结构域受体1（DDR1）。此外，我们将确定ECM-是否介导的刚度和随后在肿瘤细胞中的机械敏感信号传导促进ICC 发展。我们将测量肿瘤刚度，机械签名途径的激活和DDR1 人CCA样品中的表达并将其与临床结果相关联。刚度和DDR1在通过调节刚度和通过共同培养CAF和ICC细胞，分别缺乏COL1A1或DDR1。总之，拟议的研究将采用新颖的工具来回答该领域的一个长期问题，并可能为目标ICC- CAF串扰是这种破坏性恶性肿瘤的新型治疗策略。