Neuropathic pain is a leading cause of high impact pain, is often disabling and is poorly managed by current therapeutics. Here we focused on a unique group of neuropathic pain patients undergoing thoracic vertebrectomy where the dorsal root ganglia is removed as part of the surgery allowing for molecular characterization and identification of mechanistic drivers of neuropathic pain independently of preclinical models. Our goal was to quantify whole transcriptome RNA abundances using RNA-seq in pain-associated human dorsal root ganglia from these patients, allowing comprehensive identification of molecular changes in these samples by contrasting them with non-pain associated dorsal root ganglia. We sequenced 70 human dorsal root ganglia, and among these 50 met inclusion criteria for sufficient neuronal mRNA signal for downstream analysis. Our expression analysis revealed profound sex differences in differentially expressed genes including increase of IL1B, TNF, CXCL14, and OSM in male and including CCL1, CCL21, PENK and TLR3 in female dorsal root ganglia associated with neuropathic pain. Co-expression modules revealed enrichment in members of JUN-FOS signalling in males, and centromere protein coding genes in females. Neuro-immune signalling pathways revealed distinct cytokine signalling pathways associated with neuropathic pain in males (OSM, LIF, SOCS1) and females (CCL1, CCL19, CCL21). We validated cellular expression profiles of a subset of these findings using RNAscope in situ hybridization. Our findings give direct support for sex differences in underlying mechanisms of neuropathic pain in patient populations.

中文翻译：

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人类背根神经节的 RNA 分析揭示了促进神经性疼痛的机制的性别差异

神经性疼痛是高冲击性疼痛的主要原因，通常会致残并且目前的治疗方法管理不善。在这里，我们专注于一组独特的神经性疼痛患者，这些患者正在接受胸椎切除术，其中背根神经节作为手术的一部分被移除，从而允许独立于临床前模型的分子表征和神经性疼痛的机械驱动因素的鉴定。我们的目标是使用 RNA-seq 在这些患者的疼痛相关人类背根神经节中量化整个转录组 RNA 丰度，从而通过将这些样本与非疼痛相关背根神经节进行对比来全面识别这些样本中的分子变化。我们对 70 个人类背根神经节进行了测序，其中 50 个满足足够的神经元 mRNA 信号用于下游分析的纳入标准。我们的表达分析揭示了差异表达基因的显着性别差异，包括男性中 IL1B、TNF、CXCL14 和 OSM 的增加，以及女性背根神经节中与神经性疼痛相关的 CCL1、CCL21、PENK 和 TLR3 的增加。共表达模块揭示了男性 JUN-FOS 信号通路成员的富集，以及女性着丝粒蛋白编码基因的富集。神经免疫信号通路揭示了与男性（OSM、LIF、SOCS1）和女性（CCL1、CCL19、CCL21）神经性疼痛相关的不同细胞因子信号通路。我们使用 RNAscope 原位杂交验证了这些发现的一部分的细胞表达谱。我们的研究结果直接支持了患者群体神经性疼痛潜在机制的性别差异。