Supplementary Material for: Single cell analysis of the fate of injected oncogenic RasV12 cells in adult wild type Drosophila

We have injected dish-cultured oncogenic RasV12 cells into adult male flies and analysed by single cell transcriptomics their destiny within the host after 11 days. We identified in the pre-injection samples and in the 11-day post-injection samples in all 16 clusters of cells, of which 5 disappeared during the experiment in the host. The other cell clusters expanded and expressed genes involved in the regulation of cell cycle, metabolism and development. In addition, three clusters expressed genes related to inflammation and defense. Predominant among these were genes coding for phagocytosis and/or characteristic for plasmatocytes (the fly equivalent of macrophages). A pilot experiment indicated that injection into flies of oncogenic cells in which two of most strongly expressed genes had been previously silenced by RNA interference, resulted in a dramatic reduction of their proliferation in the host flies as compared to controls. As we have shown earlier, the proliferation of the injected oncogenic cells in the adult flies is a hallmark of the disease and induces a wave of transcriptions in the experimental flies. We hypothesise that this results in a bitter dialogue between the injected cells and the host. The experiments presented here should contribute to deciphering this dialogue.

我们将体外培养的致癌RasV12细胞（oncogenic RasV12 cells）注射至成年雄性果蝇体内，并通过单细胞转录组学（single cell transcriptomics）分析了注射11天后这些细胞在宿主体内的命运。我们在预注射样本与注射后11天的样本中，共鉴定出16个细胞簇，其中5个细胞簇在宿主体内的实验过程中消失。其余细胞簇发生扩增，并表达参与细胞周期、代谢与发育调控的基因。此外，另有3个细胞簇表达与炎症及防御相关的基因，其中占主导的基因包括编码吞噬作用相关蛋白的基因，以及浆细胞（plasmatocytes，即果蝇体内等效于巨噬细胞（macrophages）的细胞类型）的特征基因。一项预实验结果显示：相较于对照组，将此前通过RNA干扰（RNA interference）沉默了两个高表达核心基因的致癌细胞注射至果蝇体内后，这些细胞在宿主体内的增殖能力出现了显著下降。正如我们此前的研究所示，注射的致癌细胞在成年果蝇体内的增殖是该疾病的标志性特征，同时会诱导实验果蝇体内产生一波转录响应。我们推测，这一过程会在注射细胞与宿主之间引发一场激烈的互作对话；本研究呈现的实验将有助于解析这一互作机制。