Ablation of S1PR4 triggers CD8+ T cell expansion to reduce tumor growth and improve therapy. Ablation of S1PR4 triggers CD8+ T cell expansion to reduce tumor growth and improve therapy

We studied the role of S1PR4 on tumor progression using the PyMT mammary carcinoma mouse model and the AOM/DSS model for colitis-associated cancer. To gain insights of the underlying mechanism how S1PR4 ablation resulted in enhanced intratumoral CD8+ T cell abundance, we performed whole transcriptome profiling of total WT and S1PR4 KO colons subjected to the AOM/DSS model (day 84) as well as FACS-sorted CD8+ T cells from WT and S1PR4 KO tumors of PyMT mice. Next-generation mRNA sequencing, in triplicates, on a NextSeq 500 (PyMT CD8+ T cells) or a HiSeq 2000 (AOM/DSS model) high-throughput sequencer was used. Overall design: mRNA profiles of total colons of WT and S1PR4 KO mice at day 84 of the AOM/DSS model and of FACS-sorted CD8+ T cells from WT and S1PR4 KO PyMT tumors at the experimental endpoint (when one tumor reached a diameter of about 1.5 cm) were generated in triplicates.

本研究采用PyMT乳腺癌小鼠模型与偶氮甲烷/葡聚糖硫酸钠（AOM/DSS）结肠炎相关癌症模型，探究鞘氨醇1-磷酸受体4（S1PR4）在肿瘤进展中的作用。为阐明S1PR4敲除导致肿瘤内CD8+ T细胞浸润丰度升高的潜在分子机制，本研究对经AOM/DSS造模至第84天的野生型（WT）与S1PR4敲除（KO）小鼠的完整结肠组织，以及从PyMT模型小鼠野生型与S1PR4敲除肿瘤中经荧光激活细胞分选（FACS）得到的CD8+ T细胞开展了全转录组测序分析。
测序实验采用NextSeq 500（针对PyMT模型小鼠的CD8+ T细胞样本）与HiSeq 2000（针对AOM/DSS模型样本）两款高通量测序平台，所有样本均设置三个生物学重复。
实验整体设计如下：于AOM/DSS造模第84天采集的野生型与S1PR4敲除小鼠完整结肠组织样本，以及在实验终点（当单枚肿瘤直径达到约1.5 cm时）分选得到的PyMT模型小鼠野生型与S1PR4敲除肿瘤来源的CD8+ T细胞样本的mRNA表达谱，均设置三个生物学重复进行测序。