RNA sequencing analysis for mouse ovarian cancer cells and tumors response to B7-H3 (Cd276) depletion

Analysis of mouse ovarian cancer cell lines HM-1 and ID8, and HM-1 tumors inoculated to B6C3F1 mice with B7-H3 (Cd276) knockout (KO) by CRISPR-Cas9. We identified B7-H3 expression is upregulated in IFN-γ–low, PD-L1–low non-immunoreactive tumors of high grade serous ovarian cancer. We explored the influence of B7-H3 on immune evasion outside of its direct regulatory function on target cells by generating B7-H3 knockout cell lines and tumors in syngeneic mouse models. Total RNA from HM-1-B7-H3 KO cells, ID8-B7-H3 KO cells or their control cells, or intradermal tumors of HM-1-B7-H3 KO cells or their control cells at day 10 of tumor growth were extracted and used for RNA sequencing. Differential expression analysis was performed using DESeq2 (RRID: SCR_015687) between control (n = 3) and B7-H3 KO (n = 3) groups of the ID8 and HM-1 cell lines, and HM-1 intradermal tumors.

本研究针对小鼠卵巢癌细胞系HM-1、ID8，以及经CRISPR-Cas9技术敲除B7-H3（Cd276）的HM-1细胞接种至B6C3F1小鼠形成的肿瘤展开分析。本研究发现，在高级别浆液性卵巢癌的IFN-γ低表达、PD-L1低表达的非免疫反应性肿瘤中，B7-H3的表达呈上调状态。本研究通过同基因小鼠模型构建B7-H3敲除细胞系及肿瘤模型，探究了B7-H3除对靶细胞的直接调控功能外，其在肿瘤免疫逃逸中的作用。我们提取了HM-1-B7-H3敲除（KO）细胞、ID8-B7-H3敲除细胞及其对应对照细胞，或是肿瘤生长第10天时的HM-1-B7-H3敲除细胞皮下瘤及其对照瘤的总RNA，用于RNA测序。本研究使用DESeq2（RRID: SCR_015687）对ID8、HM-1细胞系以及HM-1皮下瘤的对照组（n=3）与B7-H3敲除组（n=3）进行了差异表达分析。