Immuno-genomic effects of JAK blockade in vivo

Small molecule inhibitors of JAK kinases have shown clinical effcacy in the treatment of certain autoimmune diseases. While these are known to block upstream JAK signalling events, their broader impact on the transcriptional footprint in immunocytes are unknown. Here we explore the effects of pan- and isoform-specific JAK blockade on the immuno-genomic network by genomic profiling. 6week old male C57BL/6 mice were gavaged with JAK inhibitor or vehicle for indicated treatment periods. Spleens were harvested and mechanically disrupted to prepare single cell suspensions. These were then stained in multiple surface marker panels to differentiate distinct immunocyte populations. Cells were sorted directly into TriZol. RNA was prepared in Trizol for gene expression profiling by Affymetrix Mouse Gene 1.0 ST Arrays.

JAK激酶（JAK kinases）小分子抑制剂在部分自身免疫性疾病的治疗中已展现出临床疗效。尽管已知该类抑制剂可阻断JAK信号通路的上游事件，但其对免疫细胞转录足迹的整体影响仍未明确。本研究借助基因组谱分析（genomic profiling）技术，探究泛JAK及亚型特异性JAK阻断对免疫基因组网络的影响。实验选用6周龄雄性C57BL/6小鼠，经口灌胃给予JAK抑制剂或赋形剂，给药时长依预设处理周期而定。处死小鼠后摘取脾脏，通过机械解离制备单细胞悬液；随后使用多组表面标记物对细胞进行染色，以区分不同免疫细胞群。细胞直接分选至TriZol试剂中，利用TriZol提取RNA，并通过Affymetrix小鼠基因1.0 ST芯片（Affymetrix Mouse Gene 1.0 ST Arrays）开展基因表达谱分析。