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

项目摘要 肝脏是一个重要的器官，也是唯一能够再生的实体器官。尽管如此，肝再生 在晚期肝病患者中受到损害，因此了解肝脏的机制 再生对再生医学具有重要意义。斑马鱼已经成为一种强大的 斑马鱼是研究再生的实验系统，有几种斑马鱼肝再生模型。 我建议使用以下方法的组合来彻底描述斑马鱼肝再生的过程： 传统工具和新技术。我将使用单细胞测序来鉴定所述细胞的更多标记物。 类型，发现罕见或短暂的细胞群体，重建谱系转换，并阐明转录 对伤害的反应。我将通过荧光原位杂交来验证这些数据， 转化为基因表达。我将利用一项新技术，将联合收割机单细胞测序数据与遗传谱系相结合 追踪揭示严重肝损伤后肝再生的细胞基础。这些发现将得到验证 用经典的谱系追踪技术研究肝祖细胞所需的信号通路， 再生过程中的细胞这些数据将进一步加深我们对肝脏的分子和细胞基础的理解 这有助于我们开发增强人类肝脏再生的疗法。

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.