Type I IFN Signaling Drives Responsiveness to Short Course Radiotherapy in a Murine Model of Rectal Cancer. Type I IFN Signaling Drives Responsiveness to Short Course Radiotherapy in a Murine Model of Rectal Cancer

Specific cell types from orthotopic murine rectal tumors were sequenced to explore the effect that short course radiotherapy has on the immune response. Overall design: MC38-luc tumor cells were orthotopically implanted in the rectum of C57BL6 female mice. Tumors were irradiated using a small animal irradiator from days 9-13 post tumor implantation. Tumors were treated with 5Gy radiation therapy for 5 consecutive daily fractions. Some mice were treated with anti-IFNAR blocking antibody. Tumors were harvested on day 14 and individual cell populations includining CD8 T cells (CD45+ CD8+), cancer associated fibroblasts (CD45-, CD31- Podoplanin+, PDGFRa+, Ly6c+), and tumor cells (GFP+) were sorted and processed for RNA sequencing.

本研究针对原位小鼠直肠肿瘤中的特定细胞类型开展测序，以探究短疗程放疗对免疫应答的调控作用。整体实验设计：将MC38-luc肿瘤细胞原位植入C57BL/6雌性小鼠的直肠内；于肿瘤植入后第9至13天，使用小动物辐照仪对肿瘤进行辐照，采用5Gy剂量的放射治疗，每日1次，连续照射5个分次。部分小鼠额外接受抗IFNAR阻断抗体处理。于肿瘤植入后第14天处死并获取肿瘤组织，分选得到CD8阳性T细胞（CD45+ CD8+）、肿瘤相关成纤维细胞（cancer associated fibroblasts, CD45- CD31- Podoplanin+ PDGFRα+ Ly6c+）以及GFP阳性肿瘤细胞（GFP+）等细胞群，随后对其进行RNA测序分析。