Precision Medicine Research Center, College of Medicine, The Catholic University of Korea, Seoul, South Korea; IRCGP, College of Medicine, The Catholic University of Korea, Seoul, South Korea; Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, South Korea.
Precision Medicine Research Center, College of Medicine, The Catholic University of Korea, Seoul, South Korea.
Biochem Biophys Res Commun. 2022 Jul 12;613:120-126. doi: 10.1016/j.bbrc.2022.05.003. Epub 2022 May 9.
The mechanism of melanoma metastasis is poorly understood, especially at the single-cell level. To understand the evolution from primary melanoma to metastasis, we investigated single-cell transcriptome profiles of parental B16 melanoma cells (B16F0) and its highly metastatic subclone (B16F10). Genomic alterations between cells were also analyzed by whole-exome sequencing. We identified 274 differentially expressed genes (DEGs) in B16F10, including upregulated genes related to metastasis, Lgals3, Sparc, Met, and Tmsb4x, and downregulated Mitf pathway genes, Ptgds, Cyb5a, and Cd63. The proportion of cycling cells and cells highly expressing Kdm5b was significantly high in B16F10 cells. Among the five subclusters of B16 cells (C1-5), C3/C4 clusters comprised both B16F0 and B16F10 cells and exhibited intermediate DEG patterns, whereas the C5 cluster mostly comprised B16F10 and showed typical metastatic characteristics. In trajectory analysis, the C4 cluster in B16F0, which showed unique characteristics (mainly cycling cells and upregulation of Mitf pathway genes), have transition potential to the C5 cluster (B16F10). Regarding genomic alterations, stepwise evolution with shared mutations, including Braf, Pten, and Trp53, and further specific alterations led to metastatic development. Our results provide deeper understanding of melanoma metastasis at the single-cell level, thus aiding further studies in melanoma metastasis control.
黑色素瘤转移的机制尚不清楚，尤其是在单细胞水平。为了了解从原发性黑色素瘤到转移的演变，我们研究了亲本 B16 黑色素瘤细胞（B16F0）及其高转移性亚克隆（B16F10）的单细胞转录组谱。还通过全外显子组测序分析了细胞间的基因组改变。我们在 B16F10 中鉴定出 274 个差异表达基因（DEGs），包括与转移相关的上调基因 Lgals3、Sparc、Met 和 Tmsb4x，以及下调的 Mitf 途径基因 Ptgds、Cyb5a 和 Cd63。B16F10 细胞中循环细胞和高度表达 Kdm5b 的细胞比例明显较高。在 B16 细胞的五个亚群（C1-5）中，C3/C4 亚群包含 B16F0 和 B16F10 细胞，并表现出中间的 DEG 模式，而 C5 亚群主要包含 B16F10 并表现出典型的转移性特征。在轨迹分析中，B16F0 中的 C4 亚群表现出独特的特征（主要是循环细胞和 Mitf 途径基因的上调），具有向 C5 亚群（B16F10）转变的潜力。关于基因组改变，包括 Braf、Pten 和 Trp53 在内的共享突变的逐步演变，以及进一步的特定改变导致了转移的发展。我们的研究结果在单细胞水平上提供了对黑色素瘤转移的更深入了解，从而有助于进一步研究黑色素瘤转移的控制。
