Lineage tracing and single-cell analysis reveal proliferative Prom1+tumour-propagating cells and their dynamic cellular transition during liver cancer progression
Lineage tracing and single-cell analysis reveal proliferative Prom1+tumour-propagating cells and their dynamic cellular transition during liver cancer progression
复制标题
DOI：
10.1136/gutjnl-2021-324321
复制
发表时间：
2021-09-28
期刊：
影响因子：
24.5
通讯作者：
Ma, Stephanie
中科院分区：
文献类型：
--
作者：
Zhou, Lei;Yu, Ken H. O.;Ma, Stephanie
目的肝细胞癌（HCC）具有高度的瘤内异质性，这导致了治疗耐药性和肿瘤复发。我们以前确定Prominin-1（PROM 1）/CD 133作为一个重要的肝癌干细胞（CSC）标记在人肝癌。本研究的目的是研究Prom 1+细胞在完整小鼠肝癌模型中的异质性和特性。设计我们建立了两种小鼠模型，分别代表慢性纤维化HCC和快速脂肪变性相关HCC。我们使用Prom 1（C-L/+）; Rosa 26（tdTomato/+）小鼠进行HCC诱导后谱系追踪，并使用Prom 1（C-L/+）; Rosa 26（DTA/+）小鼠进行靶向消除。进行单细胞RNA测序（scRNA-seq）以分析追踪的Prom 1+细胞的转录组学谱。结果Prom 1在HCC肿瘤中标志着具有CSC样特性的增殖性肿瘤增殖细胞。谱系追踪表明，这些细胞在原发性肿瘤中原位显示克隆扩增。标记的Prom 1+细胞在3D培养和同种异体移植中表现出增加的致瘤性，以及在移植时形成不同谱系癌症的潜力。在两种HCC模型中，Prom 1+细胞的消耗阻碍肿瘤生长并减少恶性肿瘤标志物。scRNA-seq分析强调了Prom 1 + HCC细胞的异质性，其遵循具有高增殖和干细胞特征的去分化状态的轨迹。Prom 1家系保守的基因特征预测人HCC的不良预后活性氧化解毒是去分化转变和谱系繁殖的保护机制的基础。结论本研究结合体内谱系追踪和scRNA-seq技术，揭示了Prom 1 + HCC细胞的异质性和动态变化，为深入研究恶性CSC样细胞在HCC进展中的作用机制提供了新的思路。
Objective Hepatocellular carcinoma (HCC) has high intratumoral heterogeneity, which contributes to therapeutic resistance and tumour recurrence. We previously identified Prominin-1 (PROM1)/CD133 as an important liver cancer stem cell (CSC) marker in human HCC. The aim of this study was to investigate the heterogeneity and properties of Prom1+ cells in HCC in intact mouse models. Design We established two mouse models representing chronic fibrotic HCC and rapid steatosis-related HCC. We performed lineage tracing post-HCC induction using Prom1(C-L/+); Rosa26(tdTomato/+) mice, and targeted depletion using Prom1(C-L/+); Rosa26(DTA/+) mice. Single-cell RNA sequencing (scRNA-seq) was carried out to analyse the transcriptomic profile of traced Prom1+ cells. Results Prom1 in HCC tumours marks proliferative tumour-propagating cells with CSC-like properties. Lineage tracing demonstrated that these cells display clonal expansion in situ in primary tumours. Labelled Prom1+ cells exhibit increasing tumourigenicity in 3D culture and allotransplantation, as well as potential to form cancers of differential lineages on transplantation. Depletion of Prom1+ cells impedes tumour growth and reduces malignant cancer hallmarks in both HCC models. scRNA-seq analysis highlighted the heterogeneity of Prom1+ HCC cells, which follow a trajectory to the dedifferentiated status with high proliferation and stem cells traits. Conserved gene signature of Prom1 linage predicts poor prognosis in human HCC. The activated oxidant detoxification underlies the protective mechanism of dedifferentiated transition and lineage propagation. Conclusion Our study combines in vivo lineage tracing and scRNA-seq to reveal the heterogeneity and dynamics of Prom1+ HCC cells, providing insights into the mechanistic role of malignant CSC-like cells in HCC progression.