LMO1 expression in neuroblastoma cells reprograms tumor-associated macrophages to promote metastasis.

Our earlier study has showed that the transcriptional coactivator LMO1 synergizes with the MYCN oncogene to enhance NB tumorigenesis and metastasis. To better understand how these two oncogenes cooperate during NB development, we performed single-cell RNA sequencing analysis on primary tumors from transgenic zebrafish with overexpression of MYCN, either alone or in combination with LMO1. Interestingly, we found significantly enriched signaling pathways involved in biogenesis and metabolism in NB cells with overexpression of LMO1 and MYCN. In addition, the macrophage populations from the two tumor types were distinctly clustered. Overall design: Primary neuroblastoma tumors from MYCN (n = 2) and MYCN;LMO1 (n = 3) zebrafish lines were isolated, dissociated and submitted individually for a single-cell RNA-sequencing. cDNA libraries from single cells were prepared using the SMART-seq strategy and subjected to Illumina Hi-Seq sequencing at the Harvard Medical School Biopolymers Facility. A total of 16,630 cells were sequenced, and 9,082 high-quality transcriptomes were obtained after quality control and filtering from the 5 tumor samples.

我们此前的研究已证实，转录共激活因子LMO1可与MYCN癌基因协同作用，增强神经母细胞瘤（Neuroblastoma，NB）的发生与转移能力。为进一步阐明这两种癌基因在NB发生过程中的协同调控机制，我们对仅过表达MYCN，或同时过表达MYCN与LMO1的转基因斑马鱼原代肿瘤组织开展了单细胞RNA测序分析。有趣的是，我们发现同时过表达LMO1与MYCN的NB细胞中，参与生物发生与代谢的信号通路显著富集。此外，两种肿瘤来源的巨噬细胞群呈现出显著的聚类差异。整体实验设计：我们分别分离了MYCN过表达组（n = 2）与MYCN;LMO1共过表达组（n = 3）的转基因斑马鱼原代神经母细胞瘤组织，将其解离后分别进行单细胞RNA测序。我们采用SMART-seq技术制备单细胞cDNA文库，并于哈佛医学院生物聚合物实验室开展Illumina HiSeq测序。本次测序共获得16,630个细胞的测序数据，经质量控制与过滤后，从5份肿瘤样本中得到9,082个高质量转录组数据。