Identification of novel regulators of liver regeneration using single-cell approaches

项目摘要 肝脏是人体的重要器官，也是唯一能够再生的实体器官。尽管如此，肝脏再生 在晚期肝病患者中受到损害，因此了解肝脏的机制 再生对于再生医学有着重要的意义。斑马鱼已经成为一种强大的 研究再生的实验系统，斑马鱼的肝脏再生有几种模型。 我建议使用一个组合来彻底描述斑马鱼肝脏再生的过程。 经典的工具和新的技术。我将使用单细胞测序来为所描述的细胞识别更多的标记 类型，发现稀有或短暂的细胞群，重建谱系转变，并阐明转录 对伤害的反应。我将通过多色荧光原位杂交来验证这些数据，以获得空间洞察力 转化为基因表达。我将利用一种新技术将单细胞测序数据与遗传谱系相结合 示踪揭示严重肝损伤后肝再生的细胞学基础。这些发现将得到证实 用经典的谱系示踪技术研究肝祖细胞所需的信号通路 细胞在再生过程中。这些数据将进一步加深我们对肝脏分子和细胞基础的理解。 斑马鱼的再生，有助于我们开发治疗方法来增强人类的肝脏再生。

Project Summary The liver is an essential organ and the only solid organ capable of regeneration. Despite this, liver regeneration is compromised in patients with advanced liver disease, and so understanding the mechanisms of liver regeneration has significant implications for regenerative medicine. Zebrafish have emerged as a powerful experimental system for studying regeneration, and there are several models of liver regeneration in zebrafish. I propose to thoroughly characterize the process of liver regeneration in zebrafish using a combination of classical tools and new techniques. I will use single-cell sequencing to identify more markers for described cell types, discover rare or transient cell populations, reconstruct lineage transitions, and elucidate the transcriptional responses to injury. I will validate these data by multicolor fluorescent in situ hybridization to gain spatial insight into gene expression. I will utilize a new technique to combine single-cell sequencing data with genetic lineage tracing to reveal the cellular basis of liver regeneration after severe liver injury. These findings will be validated with classical lineage tracing techniques in order to study the signaling pathways required for liver progenitor cells during regeneration. These data will further our understanding of the molecular and cellular basis of liver regeneration in zebrafish, aiding our ability to develop therapies to enhance liver regeneration in humans.