Single nuclei RNA sequencing of mouse dorsal root, nodose, and trigeminal ganglia from humanized JAK1 gain-of-function mice

Janus kinase 1 (JAK1) critically mediates downstream signaling from a variety of immune receptors. Gain-of-function mutations in JAK1 in humans lead to immune dysregulation and exaggerated type 2 immune responses, including atopic dermatitis. We have previously shown that JAK1 expressed in neurons mediates cytokine-enhanced itch signaling in atopic dermatitis. We generated a novel mouse expressing human JAK1 with the gain-of-function mutation p.A634D. To better understand how enhanced JAK1 signaling my impact sensory neurons, we performed single nuclei RNA sequencing on different sensory ganglia from JAK1 gain-of-function mice and littermate controls. Overall design: 10-12-week-old C57BL6 mice heterozygous for an allele where the mouse Jak1 gene was replaced by human JAK1 with the p.A634D mutation, or homozygous for wild type mouse Jak1, were used. Thoracic dorsal root ganglia, nodose ganglia, and trigeminal ganglia were dissociated into single nuclei preps and sequenced using the 10X Genomics multiome pipeline.

Janus激酶1（JAK1）可关键性介导多种免疫受体的下游信号转导。人类体内JAK1的功能获得性突变会引发免疫失调以及过度的2型免疫应答，其中包括特应性皮炎。我们此前的研究证实，神经元中表达的JAK1可介导特应性皮炎中细胞因子增强的瘙痒信号通路。本研究构建了一株携带功能获得性突变p.A634D的人源JAK1转基因小鼠。为深入解析增强的JAK1信号转导对感觉神经元的影响，我们对JAK1功能获得性突变小鼠及其同窝野生型对照小鼠的不同感觉神经节开展了单细胞核RNA测序。实验整体设计：实验对象为10至12周龄的C57BL/6品系小鼠，分为两组：一组为将小鼠内源性Jak1基因替换为携带p.A634D突变的人源JAK1的杂合子小鼠，另一组为小鼠Jak1基因野生型纯合的同窝对照。研究人员将胸椎背根神经节、结状神经节与三叉神经节解离为单细胞核悬液，采用10X Genomics多组学测序流程完成测序。