Identifying therapeutic options for intrahepatic cholangiocarcinoma

确定肝内胆管癌的治疗选择

• 批准号: 10392447
• 负责人: Sungjin Ko
• 金额: $ 32.22万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10392447
• 关键词: Address; Automobile Driving; Azacitidine; Biliary; Binding; Bioinformatics; Biological Markers; Cell Differentiation process; Cells; ChIP-seq; Cholangiocarcinoma; Clinic; Clinical; Data; Data Set; Development; Disease; Doxycycline; Epigenetic Process; Epithelial Cells; Excision; Exposure to; Gene Expression; Genes; Genetic; Genetic Transcription; Genome; Hepatocyte; Heterogeneity; Human; Injury; Intrahepatic Cholangiocarcinoma; Lead; Liver; Malignant neoplasm of liver; Mediating; Medical; Modeling; Molecular; Molecular Profiling; Mus; Newly Diagnosed; Oncogenes; Operative Surgical Procedures; Pathogenesis; Pathologic; Patients; Pharmaceutical Preparations; Pharmacology; Phosphorylation Site; Plasmids; Play; Proto-Oncogene Proteins c-akt; Repression; Risk; Role; Signal Transduction; Site; Sleeping Beauty; Survival Rate; System; Tamoxifen; Testing; Therapeutic; Therapeutic Effect; Thioacetamide; Time; Toxin; Unresectable; Zebrafish; alpha Toxin; base; bioinformatics tool; bisulfite sequencing; chronic liver disease; clinically relevant; cohort; genetic signature; in vivo; inhibitor; innovation; methylome; mutant; neoplastic cell; new therapeutic target; notch protein; novel therapeutics; pre-clinical; precision medicine; prevent; progenitor; response; small molecule; targeted treatment; therapeutic evaluation; therapeutic target; tool; toxicant; transcriptome sequencing; tumor; tumor growth; tumor heterogeneity; tumor initiation; tumorigenesis; Address; Automobile Driving; Azacitidine; Biliary; Binding; Bioinformatics; Biological Markers; Cell Differentiation process; Cells; ChIP-seq; Cholangiocarcinoma; Clinic; Clinical; Data; Data Set; Development; Disease; Doxycycline; Epigenetic Process; Epithelial Cells; Excision; Exposure to; Gene Expression; Genes; Genetic; Genetic Transcription; Genome; Hepatocyte; Heterogeneity; Human; Injury; Intrahepatic Cholangiocarcinoma; Lead; Liver; Malignant neoplasm of liver; Mediating; Medical; Modeling; Molecular; Molecular Profiling; Mus; Newly Diagnosed; Oncogenes; Operative Surgical Procedures; Pathogenesis; Pathologic; Patients; Pharmaceutical Preparations; Pharmacology; Phosphorylation Site; Plasmids; Play; Proto-Oncogene Proteins c-akt; Repression; Risk; Role; Signal Transduction; Site; Sleeping Beauty; Survival Rate; System; Tamoxifen; Testing; Therapeutic; Therapeutic Effect; Thioacetamide; Time; Toxin; Unresectable; Zebrafish; alpha Toxin; base; bioinformatics tool; bisulfite sequencing; chronic liver disease; clinically relevant; cohort; genetic signature; in vivo; inhibitor; innovation; methylome; mutant; neoplastic cell; new therapeutic target; notch protein; novel therapeutics; pre-clinical; precision medicine; prevent; progenitor; response; small molecule; targeted treatment; therapeutic evaluation; therapeutic target; tool; toxicant; transcriptome sequencing; tumor; tumor growth; tumor heterogeneity; tumor initiation; tumorigenesis

In the US, approximately 10,000 patients are newly diagnosed annually with cholangiocarcinoma, and their 5-year survival rate is less than 10%. Heterogeneity in cellular origins and molecular signatures of intrahepatic cholangiocarcinoma (ICC) highlight the demand for biomarkers to stratify tumors and generate targeted therapies. Besides biliary epithelial cells (BECs), hepatocytes (HCs) have been considered as a cellular origin of human ICC. This is evident by rapid HC-driven ICC induction through the co-expression of activated AKT (myrAKT) and Notch intracellular domain (NICD) in mouse HCs using sleeping beauty transposon system. Our preliminary analysis using large ICC patient cohorts demonstrates that the genetic signature of AKT-NICD (AN)-driven murine ICC correlates significantly with ~30% of human ICC, supporting the clinical relevance of this ICC model. Deletion of either Sox9 or Yap delayed ICC formation, and instead induced AN-driven SOX9-/YAP+ or SOX9+/YAP- ICC respectively, which also have been identified in human ICC. These data indicate that Notch independently regulates Sox9 and Yap in HC-driven ICC. Importantly, we found that co-repression of Yap and Sox9 completely prevents Notch- dependent ICC formation. However, the mechanisms of tumorigenesis driven by YAP and SOX9, as well as their interactions with AKT signaling, remain poorly understood. Based on our preliminary observations, our central hypothesis is that HC-driven ICC tumor growth depends on transcriptional and epigenetic alterations driven by two major downstream effectors of Notch signaling, SOX9 and YAP, alongside AKT signaling. In aim 1, we aim to test therapeutic effect of co-repression of YAP and SOX9 in advanced ICCs to better address the clinical need for late-stage therapy. We will employ genetic AN-ICC model as well as liver toxin-based ICC model using innovative inducible gene modulation systems to induce simultaneous, conditional and inducible Sox9 and YAP repression in advanced ICC. In aim 2, we are proposing 3 subaims to delineate the molecular mechanisms underlying AN- mediated HC-driven ICC formation. First, using ChIP-seq and bioinformatic tools, we will identify both the unique and the overlapping downstream targets regulated by Sox9 and Yap during HC-derived ICC formation. Second, we will elucidate the pathologic role of the NICD-YAP/TEAD-DNMT1 epigenetic axis in HC-driven ICC development by modulating this axis with pharmacological and genetic tools in the in vivo system and studying its effects on the methylome of ICC tumors. Pursuing these 2 subaims, we will identify downstream effectors of Sox9 and Yap for more selective and safer therapeutic targeting in ICC. Third, we aim to elucidate how AKT mediates HC-to-ICC transformation, which remains poorly understood. The successful execution of this study will 1) provide an essential evidence for considering SOX9 and YAP co- inhibition as an attractive candidate for precision medicine therapy, which is theorized to be the most effective approach to conquer lethal tumor, and 2) reveal the key downstream effectors for AKT, SOX9 and YAP which will help to develop more potent therapeutic options for ICC.

在美国，每年大约有10,000名患者每年被新诊断出患有胆管癌及其5年 生存率小于10％。肝内胆管癌的细胞起源和分子特征的异质性 （ICC）强调了对生物标志物对肿瘤进行分层并产生靶向疗法的需求。除了胆道上皮细胞 （BEC），肝细胞（HCS）被认为是人类ICC的细胞起源。快速HC驱动可以看出这一点 ICC通过激活的Akt（Myrakt）和小鼠的凹口内结构域（NICD）的共表达诱导 HC使用睡美人转座系统。 我们使用大型ICC患者队列的初步分析表明，Akt-Nicd的遗传特征 （AN）驱动的鼠ICC与〜30％的人类ICC显着相关，支持该ICC的临床相关性 模型。删除SOX9或YAP延迟的ICC形成，而是诱导AN驱动的Sox9-/Yap+或Sox9+/Yap- ICC分别在人类ICC中也已确定。这些数据表明Notch独立调节 HC驱动的ICC中的SOX9和YAP。重要的是，我们发现YAP和SOX9的共抑制完全阻止了Notch- 依赖的ICC形成。但是，由YAP和SOX9驱动的肿瘤发生机制及其 与AKT信号的相互作用，仍然了解不足。基于我们的初步观察，我们的中心假设 是HC驱动的ICC肿瘤的生长取决于两个主要下游驱动的转录和表观遗传变化 Notch信号，SOX9和YAP的效应器以及AKT信号传导。 在AIM 1中，我们旨在测试YAP和SOX9在高级ICC中的治疗效果以更好地解决 晚期治疗的临床需求。我们将采用遗传AN-ICC模型以及基于肝毒素的ICC模型 使用创新的诱导基因调制系统来诱导同时，条件和诱导的Sox9和Yap 高级ICC中的抑制。在AIM 2中，我们提出了3个subiams来描述An-的分子机制 介导的HC驱动的ICC组。首先，使用Chip-seq和生物信息学工具，我们将确定唯一和 在HC衍生的ICC形成过程中，由SOX9和YAP调节的下游目标重叠。其次，我们将阐明 NICD-YAP/TEAD-DNMT1表观遗传轴在HC驱动的ICC开发中的病理作用通过调节 带有药理学和遗传工具的轴，在体内系统中，研究其对ICC肿瘤甲基甲基的影响。 追求这两个子iaM，我们将确定Sox9的下游效应子和YAP的下游效应器，以获得更有选择性和更安全的治疗方法 靶向ICC。第三，我们旨在阐明AKT如何介导HC-to-ICC转化，但保持较差 理解。 这项研究的成功执行将是1）为考虑SOX9和YAP共同的基本证据提供了一个基本证据。 抑制作用是精密医学疗法的有吸引力的候选人，这是理论上是最有效的方法 征服致命肿瘤，2）揭示Akt，Sox9和Yap的关键下游效应子，这将有助于发展 ICC的更有效的治疗选择。