Mechanistic underpinnings of chronic low back pain

慢性腰痛（CLBP）是全球残疾的主要原因，目前的治疗方法主要是 无效。拟议的项目旨在使用RNA-seq技术对神经，关节，肌肉， 和筋膜组织从完全表型化的个体与CLBP进行脊柱手术，以获得新的 深入了解CLBP。我们的中心假设是神经和病理组织中的转录变化驱动了 人CLBP。我们将通过将单细胞RNA-seq应用于组织样本来测试这一假设。 对接受手术的CLBP患者进行了表征。在我们的第一个目标中，我们将应用单核和大量RNA- seq到上述外周和神经组织。我们假设，外周和神经组织从 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，以鉴定配体- 与CLBP和神经元转录组相关的细胞/组织之间的受体相互作用。我们将 从器官捐献者和慢性病患者的集体DRG数据集组装的相关伤害感受器表型 疼痛患者从CLBP组织中创建相互作用组谱，并将这些相互作用组与疼痛相关联 表型最后，在我们的第三个目标中，我们将对来自人DRG神经元的人DRG神经元进行体外药理学研究。 器官捐赠者了解假定的疼痛介质如何作用于伤害感受器。该项目将导致一个新的 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 完全基于使用人类样本和组织，可以开发用于发现、目标识别和 其他疼痛疾病的临床验证。

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.