Mechanisms of extrahepatic biliary proliferation and regeneration

肝外胆管增殖和再生的机制

基本信息

批准号：
10630123
负责人：
Nataliya Razumilava
金额：
$ 16.74万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-15 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10630123
关键词：
Acceleration Affect Attenuated Biliary Binding Biology CD44 gene Cell Communication Cells Cholangiocarcinoma Coculture Techniques Data Development Disease Epithelial Cells Epithelium Erinaceidae Extrahepatic Extrahepatic Bile Ducts Fibroblasts Flow Cytometry Funding GLI gene GLI2 gene GLI3 gene Genes Genetic Genetic Transcription Glioma Goals Health Hepatobiliary Homeostasis Human Immunohistochemistry In Situ Hybridization In Vitro Injury Ligands Ligation Mediator Mesenchymal Mission Modeling Molecular Mus Natural regeneration Organoids Pathway interactions Patients Pharmacological Treatment Play Population Production Proliferating Public Health Quality of life Regenerative Medicine Reporter Reporter Genes Research Research Personnel Role Signal Pathway Signal Transduction Source Stromal Cells System Techniques Testing Transgenic Mice Transgenic Organisms United States United States National Institutes of Health WNT Signaling Pathway beta catenin bile duct cholangiocyte effective therapy improved improved outcome in vivo inhibitor liver transplantation loss of function mouse model novel novel therapeutic intervention pharmacologic programs receptor response response to injury skills smoothened signaling pathway therapy development tool translational study

项目摘要

PROJECT SUMMARY/ABSTRACT Cholangiopathies are incurable, progressive extrahepatic bile duct (EHBD) disorders characterized by injury- induced cholangiocyte hyperproliferation. Development of therapies for cholangiopathies requires better understanding of cellular and molecular mechanisms regulating EHBD proliferation during homeostasis and in response to injury. Molecular pathways that regulate cholangiocyte proliferation are currently not well understood. This application aims to answer a fundamental question about mechanisms of EHBD regeneration and explore strategies to modulate the proliferative potential of the biliary epithelium. The rationale for the proposed research is that defining cellular and molecular interactions underlying cholangiocyte responses to injury can offer novel therapeutic strategies for cholangiopathies and hepatobiliary regenerative medicine. Our preliminary data suggest that Hedgehog (HH) and WNT signaling play important roles in EHBD homeostasis and proliferation after injury. The overarching hypothesis for this proposal is that HH and WNT signaling regulate crosstalk between epithelial and stromal cells to promote EHBD proliferation after injury. In this proposal, we will use genetic mouse models and pharmacological treatments in vivo. Bile duct ligation will be used as an injury model. Human and mouse EHBD organoid (BDO) co-culture with primary mesenchymal cells will be used to directly study epithelial-stromal crosstalk and pathway interaction in vitro and enhance the translational component of this project. Aim 1 will focus on HH signaling and determine if the Indian HH ligand from cholangiocytes signals to GLI1+ HH-responsive fibroblasts to indirectly regulate cholangiocyte proliferation. Aim 2 will focus on WNT signaling and test if WNT from GLI1-expresing cells is critical for cholangiocyte proliferation. Aim 3 will focus on cell-cell and pathway interactions and determine if IHH from cholangiocytes directly regulates WNT production by fibroblasts to induce cholangiocyte proliferation. Under Aims 1 and 2 we will use transgenic reporter and loss-of-function mouse models, pharmacological inhibitors of HH and WNT signaling, in situ hybridization, and immunohistochemistry techniques to define the in vivo effects. Under Aim 3 we take a reductionist approach by using organoid co-culture models. It will also support the development of robust new tools (transgenic mice and organoid models) and skills (in situ hybridization and flow cytometry) to support an independent research program focused on fundamental signaling pathways regulating EHBD. This new K08 application will also promote the development of the PI into an independent NIH-funded investigator and support her long-term goal to understand fundamental mechanisms of EHBD biology to ultimately improve outcomes in patients with cholangiopathies.

项目概要/摘要胆管病是无法治愈的进行性肝外胆管 (EHBD) 疾病，其特征是损伤诱导胆管细胞过度增殖。胆管病治疗方法的开发需要更好的了解体内平衡过程中调节 EHBD 增殖的细胞和分子机制对伤害的反应。调节胆管细胞增殖的分子途径目前尚不完善明白了。该应用旨在回答有关 EHBD 再生机制的基本问题并探索调节胆管上皮增殖潜力的策略。理由如下：拟议的研究是定义胆管细胞反应背后的细胞和分子相互作用损伤可以为胆管病和肝胆再生医学提供新的治疗策略。我们的初步数据表明 Hedgehog (HH) 和 WNT 信号在 EHBD 稳态中发挥重要作用以及损伤后的增殖。该提案的总体假设是 HH 和 WNT 信号传导调节上皮细胞和基质细胞之间的串扰，促进损伤后 EHBD 增殖。在这个建议中，我们将使用基因小鼠模型和体内药物治疗。胆管结扎会用作损伤模型。人和小鼠 EHBD 类器官 (BDO) 与原代间充质共培养细胞将用于直接研究体外上皮基质串扰和通路相互作用，并增强该项目的翻译部分。目标 1 将重点关注 HH 信号传导并确定印度 HH 配体是否从胆管细胞向 GLI1+ HH 反应性成纤维细胞发出信号，间接调节胆管细胞增殖。目标 2 将重点关注 WNT 信号传导并测试来自 GLI1 表达细胞的 WNT 是否对于胆管细胞增殖。目标 3 将重点关注细胞与细胞和通路的相互作用，并确定 IHH 是否来自胆管细胞直接调节成纤维细胞产生WNT以诱导胆管细胞增殖。在下面目标 1 和 2 我们将使用转基因报告基因和功能丧失小鼠模型、药物抑制剂 HH 和 WNT 信号传导、原位杂交和免疫组织化学技术来定义体内影响。在目标 3 下，我们通过使用类器官共培养模型采取还原论方法。它还将支持开发强大的新工具（转基因小鼠和类器官模型）和技能（原位杂交和流式细胞术）以支持专注于基本信号通路的独立研究计划调节 EHBD。这个新的K08应用也将推动PI发展成为独立的 NIH 资助的研究人员并支持她了解 EHBD 基本机制的长期目标生物学最终改善胆管病患者的预后。