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.

项目概要 肝脏表现出卓越的再生能力，但慢性损伤或严重急性损伤可能会导致肝脏损伤。 破坏再生机制并导致终末期肝病。缺乏有效的药物治疗 加上供体器官可用性不足，需要开发新的再生医学 - 基础疗法和对肝脏修复基本机制的更深入了解。转录因子 Sox9 是许多上皮组织中干细胞/祖细胞功能所必需的，并且已被证明 建立细胞身份。在肝脏中，Sox9 对于胆道上皮细胞 (BEC) 规范的计时是必需的 发育并在成人 BEC 中广泛表达。 Sox9 缺乏会加重小鼠模型中的胆管病 阿拉吉尔综合症。我的初步数据表明 Sox9 敲除小鼠的导管缺乏，这表明 Sox9 在 BEC 规范中的核心作用。该提案旨在定义肝内 BEC 的功能调节 在发育和损伤相关的导管反应 (DR) 过程中，由 Sox9 形成的网络。 DR是一种常见的 肝脏疾病的特征，由 BEC 的增殖性扩张来定义。此外，基于 损伤类型、时间和/或损伤程度，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 的基因调控仍知之甚少。这 该提案的中心假设是需要 Sox9 来建立适当的肝内 BEC 发育和导管反应中的网络。我将通过以下具体目标来检验这个假设： 目标 1A 将确定 Sox9 在小鼠中形成功能性肝内胆管的发育要求 通过使用组织学、胆汁酸测定、类器官测定和 3D 组织成像。目标 1B 将进行调查 Sox9 在通过 scRNA-seq 指定 BEC 亚群中的作用。目标 2 将决定 Sox9 如何维持 使用可诱导的 Sox9 敲除小鼠在 DR 期间观察成年 BEC 群体。通过肝脏检测BEC功能 血清生物化学、组织学和谱系追踪。该项目将决定 Sox9 如何建立和 维持 BEC 人口并为我提供培训，以实现我获得专注于研究的教师的目标 研究胃肠道组织稳态和再生的职位。这项工作意义重大，因为研究结果将有助于 了解肝脏修复的基本机制，可用于确定肝脏修复的新靶点 恢复终末期肝病肝功能的疗法。

项目成果

Tunable fluorescent probes for detecting aldehydes in living systems.

用于检测生命系统中醛类的可调谐荧光探针。

• 发表时间: 2024-03-27
• 影响因子: 8.4
• 作者: Wills, Rachel;Shirke, Rajendra;Hrncir, Hannah;Talbott, John M;Sad, Kirti;Spangle, Jennifer M;Gracz, Adam D;Raj, Monika
• 通讯作者: Raj, Monika