Identifying therapeutic options for intrahepatic cholangiocarcinoma

确定肝内胆管癌的治疗选择

基本信息

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
项目类别：
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

项目摘要

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，每年约有 10,000 名患者新诊断出胆管癌，他们的 5 年病程存活率低于10%。肝内胆管癌细胞起源和分子特征的异质性（ICC）强调了对生物标志物的需求，以对肿瘤进行分层并产生靶向治疗。除胆管上皮细胞外 (BEC)、肝细胞(HC)被认为是人类ICC的细胞起源。这通过快速 HC 驱动即可明显看出通过在小鼠中共表达激活的 AKT (myrAKT) 和 Notch 胞内结构域 (NICD) 诱导 ICC 使用睡美人转座子系统的HC。我们使用大型 ICC 患者队列进行的初步分析表明，AKT-NICD 的遗传特征 (AN) 驱动的小鼠 ICC 与约 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 会延迟 ICC 形成，并诱导 AN 驱动的 SOX9-/YAP+ 或 SOX9+/YAP- ICC 分别在人类 ICC 中得到鉴定。这些数据表明Notch独立监管 Sox9 和 Yap 在 HC 驱动的 ICC 中。重要的是，我们发现 Yap 和 Sox9 的共同抑制完全阻止了 Notch- 依赖ICC的形成。然而，YAP和SOX9驱动肿瘤发生的机制及其作用与 AKT 信号传导的相互作用仍然知之甚少。根据我们的初步观察，我们的中心假设 HC 驱动的 ICC 肿瘤生长取决于两个主要下游驱动的转录和表观遗传改变 Notch 信号传导、SOX9 和 YAP 以及 AKT 信号传导的效应器。在目标 1 中，我们的目标是测试 YAP 和 SOX9 共抑制在晚期 ICC 中的治疗效果，以更好地解决临床需要晚期治疗。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诱导同步、条件和诱导型 Sox9 和 YAP 高级 ICC 中的镇压。在目标 2 中，我们提出了 3 个子目标来描述 AN- 的分子机制介导 HC 驱动的 ICC 形成。首先，使用 ChIP-seq 和生物信息学工具，我们将识别独特的和在 HC 衍生的 ICC 形成过程中，Sox9 和 Yap 调节的下游目标重叠。其次，我们要阐明 NICD-YAP/TEAD-DNMT1 表观遗传轴在 HC 驱动的 ICC 发育中的病理作用轴与体内系统中的药理学和遗传工具，并研究其对 ICC 肿瘤甲基化组的影响。追求这两个子目标，我们将确定 Sox9 和 Yap 的下游效应器，以实现更具选择性和更安全的治疗 ICC 中的目标。第三，我们的目标是阐明 AKT 如何介导 HC 到 ICC 的转化，这一转化仍然很差明白了。这项研究的成功执行将 1) 为考虑 SOX9 和 YAP 共同提供重要证据抑制作为精准医学治疗的一个有吸引力的候选者，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，这将有助于开发 ICC 更有效的治疗选择。