Establishing Sox9 as a regulator of intrahepatic bile duct development and regeneration

建立 Sox9 作为肝内胆管发育和再生的调节因子

• 批准号: 10656256
• 负责人: Hannah Rose Hrncir
• 金额: $ 4.77万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656256
• 关键词: 3-Dimensional; Ablation; Acute; Adult; Alagille Syndrome; Alternative Therapies; Apoptosis; Bile Acids; Bile fluid; Biliary; Biliary Atresia; Biochemistry; Biological Assay; Biology; Cell Differentiation process; Cell Maintenance; Cell physiology; Cellular Morphology; Characteristics; Chronic; Combined Modality Therapy; Data; Development; Duct (organ) structure; Epithelial Cells; Epithelium; Exhibits; Faculty; Failure; Fibrosis; Genetic; Genetic Transcription; Goals; Hepatocyte; Heterogeneity; Histology; Homeostasis; Immunohistochemistry; Impairment; Injury; Intrahepatic bile duct; Knock-out; Knockout Mice; Liver; Liver Regeneration; Liver diseases; Mediating; Medical; Morphogenesis; Morphology; Mus; Natural regeneration; Organ Donor; Organoids; Patient-Focused Outcomes; Patients; Phenotype; Play; Population; Positioning Attribute; Prevalence; Proliferating; Reaction; Recovery; Regenerative Medicine; Regenerative capacity; Regulation; Reporting; Research; Risk; Role; Secure; Serum; Specific qualifier value; Testing; Tissue imaging; Tissues; Training; Transcriptional Regulation; Transgenes; Transplantation; United States; Work; bile duct; conditional knockout; end stage liver disease; experimental study; graft failure; improved; intrahepatic; liver function; liver injury; liver repair; liver transplantation; mouse model; new therapeutic target; novel; regenerative; single-cell RNA sequencing; stem cell function; tissue regeneration; transcription factor

PROJECT SUMMARY The liver exhibits remarkable capacity for regeneration, but chronic injury or severe acute damage can overwhelm regenerative mechanisms and lead to end-stage liver disease. The lack of effective medical therapies combined with insufficient donor organ availability necessitates the development of new regenerative medicine- based therapies and a deeper understanding of the basic mechanisms of liver repair. The transcription factor Sox9 is required for stem/progenitor cell function in a number of epithelial tissues and has been shown to establish cellular identity. In the liver, Sox9 is required for timing of biliary epithelial cell (BEC) specification in development and is broadly expressed in adult BECs. Sox9 deficiency worsens cholangiopathy in mouse models of Alagille Syndrome. My preliminary data demonstrate ductal paucity in Sox9 knockout mice, suggesting a central role for Sox9 in BEC specification. This proposal seeks to define functional regulation of intrahepatic BEC networks by Sox9, during development and injury-associated ductular reaction (DR). DR is a common characteristic of liver disease and is defined by the proliferative expansion of BECs. Furthermore, based on damage type, timing, and/or extent of damage, DR can involve lineage conversion between mature hepatocytes and BECs that contributes to tissue regeneration. The genetic regulation of DR remains poorly understood. The central hypothesis of this proposal is that Sox9 is required to establish proper intrahepatic BEC networks in development and ductular reaction. I will test this hypothesis with the following specific aims: Aim 1A will establish the developmental requirement of Sox9 to form functional intrahepatic bile ducts in mice through the use of histology, bile acid assays, organoid assays, and 3D tissue imaging. Aim 1B will investigate the role of Sox9 in specifying BEC subpopulations through scRNA-seq. Aim 2 will determine how Sox9 maintains adult BEC populations during DR using inducible Sox9 knockout mice. BEC function will be assayed by liver serum biochemistry, histology, and lineage tracing. This project will determine how Sox9 establishes and maintains BEC populations and provide me with training towards my goal of obtaining a research-focused faculty position studying GI tissue homeostasis and regeneration. This work is significant because the findings will help develop an understanding of basic mechanisms of liver repair that can be used to identify novel targets for therapeutics to restore liver function in end stage liver disease.

项目摘要 肝脏具有显着的再生能力，但是慢性损伤或严重急性损害可以 压倒性的再生机制并导致终末期肝病。缺乏有效的医疗疗法 结合不足的供体器官可用性，需要开发新的再生医学 - 基于疗法和对肝修复基本机制的更深入了解。转录因子 Sox9是许多上皮组织中的茎/祖细胞功能所必需的，并且已证明是 建立细胞身份。在肝脏中，Sox9是胆汁上皮细胞（BEC）规范所必需的 发展，并在成人BEC中广泛表达。 SOX9缺乏症使鼠标造成胆管病恶化 阿拉吉综合症。我的初步数据表明Sox9敲除小鼠的导管缺乏，这表明 Sox9在BEC规范中的中心作用。该提案旨在定义肝内BEC的功能调节 Sox9网络在发育和损伤相关的导管反应（DR）期间。 DR是一个普遍的 肝病的特征，由BEC的增殖膨胀来定义。此外，基于 损害类型，时机和/或损害程度，DR可能涉及成熟肝细胞之间的谱系转换 和有助于组织再生的BEC。 DR的遗传调节仍然知之甚少。这 该提议的中心假设是Sox9需要建立适当的肝内BEC 发育和导管反应的网络。我将以以下特定目的检验该假设： AIM 1A将确定SOX9的发育需求，以形成小鼠肝内胆管的功能 通过使用组织学，胆汁酸测定，器官测定和3D组织成像。 AIM 1B将调查 SOX9在通过SCRNA-SEQ指定BEC亚群中的作用。 AIM 2将决定Sox9如何维持 成年人在DR期间使用诱导型SOX9敲除小鼠的种群。 BEC功能将由肝脏测定 血清生物化学，组织学和谱系跟踪。该项目将决定Sox9如何建立和 维护人口，并为我提供培训，以获得以研究为中心的教师 研究胃肠道组织稳态和再生的位置。这项工作很重要，因为这些发现将有所帮助 对肝修复的基本机制有了了解，可用于识别新的目标 在末期肝病中恢复肝功能的疗法。