Mechanistic underpinnings of chronic low back pain

慢性下腰痛(CLBP)是全世界导致残疾的主要原因，目前的治疗方法大多是 效果不佳。拟议的项目旨在使用rna-seq技术，对神经、关节、肌肉、 和筋膜组织，来自完全表型的CLBP患者，接受脊柱手术以获得新的 洞察CLBP。我们的中心假设是神经和病理组织的转录变化驱动 人类的CLBP。我们将通过将单细胞rna-seq应用于彻底的组织样本来检验这一假设。 描述了正在接受手术的CLBP患者的特征。在我们的第一个目标中，我们将应用单核和整体RNA- 对上述外周和神经组织进行SEQ。我们假设外周和神经组织来自 CLBP患者将显示细胞类型和配体的变化，从而深入了解CLBP的不同机制。 在我们的第二个目标中，我们将把我们的计算相互作用组模型应用于我们的RNA-SEQ数据，以确定配体-- CLBP相关细胞/组织与神经元转录本之间的受体相互作用。我们会 相关伤害性感受器表型来自我们收集的来自器官捐赠者和慢性病患者的DRG数据集 疼痛患者从CLBP组织中创建交互作用组，并将这些交互作用与疼痛联系起来 表型。最后，在我们的第三个目标中，我们将对人背根神经节神经元进行体外药理学研究 器官捐赠者了解可能的疼痛调节剂如何作用于伤害性感受器。这个项目将带来一个新的 了解CLBP，并阐明完全基于疼痛表型的特定治疗机会 人类分子神经科学。该研究项目将作为一个例子，说明基于 完全依赖于使用人体样本和组织可以被开发用于发现、目标识别和 其他疼痛障碍的临床验证。

Chronic low back pain (CLBP) is the leading cause of disability worldwide and current treatments are mostly ineffective. The proposed project aims to use RNA-seq techniques with cellular resolution on nerve, joint, muscle, and fascia tissues from thoroughly-phenotyped individuals with CLBP undergoing spine surgery to gain new insight into CLBP. Our central hypothesis is that transcriptional changes in nerves and pathological tissues drive human CLBP. We will test this hypothesis by applying single cell RNA-seq to tissue samples of thoroughly characterized patients with CLBP undergoing surgery. In our first aim we will apply single nucleus and bulk RNA- seq to the peripheral and neural tissues outlined above. We hypothesize that peripheral and neural tissues from patients with CLBP will display shifts in cell types and ligands that give insight into distinct mechanisms of CLBP. In our second aim we will apply our computational interactome model to our RNA-seq data to identify ligand- receptor interactions between cells/tissues associated with CLBP and neuronal transcriptomes. We will associate nociceptor phenotypes assembled from our collective DRG datasets from organ donors and chronic pain patients to create interactome profiles from CLBP tissues, and associate these interactomes to pain phenotypes. Finally, in our third aim we will conduct in vitro pharmacology studies on human DRG neurons from organ donors to understand how putative pain mediators act on nociceptors. The project will lead to a new understanding of CLBP, and elucidate pain phenotype-specific therapeutic opportunities based entirely on human molecular neuroscience. The research project will serve as an example of how platforms that are based entirely on the use of human samples and tissues can be developed for discovery, target identification and clinical validation for other pain disorders.