Gene and microRNA expression data from tumor induced CD11b+ MDSC. Mus musculus

Tumor growth is associated with a profound alteration of myelopoiesis, leading to recruitment of immunosuppressive cells known as myeloid-derived suppressor cells (MDSCs). Immuno-regulatory activity of both tumor-induced and BM-derived MDSCs (by GM-CSF and IL-6 treatment) was entirely dependent on C/EBP transcription factor (TF), a key component of the emergency myelopoiesis triggered by stress and inflammation. We used miR expression analysis to identify miRs which could drive MDSC recruitment/generation/activity by modulating specific TFs and pathway. In particular, we identified a miR signature of 79 miR differentially expressed between not suppressive CD11b+ cells and CD11b+ isolated from tumor mass and spleen of tumor-bearing mice. Moreover on the same samples we profiled gene expression with Affymetrix microarrays to perform an integrated analysis of mirna and gene expression. Keywords: Expression profiling by array Overall design: CD11b+ cells were immuno-magnetically enriched from various mouse tissues, and total RNA isolated by Trizol reagent. CD11b+ cells obtained from the spleen of healthy BALB/c and C57BL/6 mice were used as reference sample for tumor induced CD11b+ MDSC, enriched from either the spleen and the tumor infiltrate of tumor-bearing mice. A triplicate of each sample was considered (except for EL4 samples, where we considered a duplicated), and each sample derives from a pool of three mice, at least.

肿瘤生长与骨髓生成的显著改变密切相关，可招募被称为髓系来源抑制细胞（myeloid-derived suppressor cells, MDSCs）的免疫抑制性细胞。经粒细胞-巨噬细胞集落刺激因子（GM-CSF）与白细胞介素6（IL-6）处理后，肿瘤诱导及骨髓来源的MDSCs的免疫调节活性，完全依赖于CCAAT增强子结合蛋白（C/EBP）转录因子（transcription factor, TF）——该因子是应激与炎症触发的紧急骨髓生成的关键组分。我们通过microRNA（miR）表达分析，筛选可通过调控特定转录因子与通路来介导MDSC招募、生成及活化的miR。具体而言，我们鉴定出一组由79个miR组成的特征谱，这些miR在非抑制性CD11b+细胞与从荷瘤小鼠肿瘤组织及脾脏中分离的CD11b+细胞之间存在差异表达。此外，我们利用Affymetrix基因芯片对同一批样本进行基因表达谱分析，以实现miRNA与基因表达的整合分析。
关键词：基于芯片的表达谱分析
实验设计：从不同小鼠组织中免疫磁珠富集CD11b+细胞，采用Trizol试剂提取总RNA。以健康BALB/c及C57BL/6小鼠脾脏来源的CD11b+细胞作为参考样本，用于比对从荷瘤小鼠脾脏及肿瘤浸润组织中富集的肿瘤诱导型CD11b+ MDSCs。每个样本设置三次生物学重复（EL4样本除外，仅设置两次重复），且每个样本至少来源于3只小鼠的混合组织。