Mapping the human DRG and spinal cord functional genome at cellular and spatial resolution

老鼠和人类的伤害性感受器之间存在着明显的差异，这些差异对 以周围神经系统为靶点的疼痛疗法的发展。我们的空间测序实验 揭示了人类背根神经节(DRG)中一组不同的细胞类型。在这个研究项目中，我们将 比较来自器官捐赠者和患有慢性疼痛的外科患者的DRG组织。我们将包括一系列年龄 在不同种族的人群中，无论是男性还是女性，在成年后的整个生命周期中。除了DRG，我们还 还将检查脊髓。我们已经进行了序列分裂和空间转录分析 背角和腹角来自选定数量的器官捐赠者，证明了这两种技术的可行性。我们的 数据显示，像背根节一样，脊髓有明显的物种差异，突显了重要性 这一发现的成果。在我们的第一个目标中，我们将使用空间排序和拆分序列技术来精确地 定义人背根神经节的细胞类型。我们将使用器官捐赠者和疼痛患者的DRG来定义细胞 转录本随着疼痛而变化，如果这种变化随性别和年龄的变化而变化的话。序列号ATAC-SEQ将识别 人背根神经节的转录状态和调控元件。在我们的第二个目标中，我们将使用相同的 绘制人类背角细胞类型图的技术。我们将努力使用DRG和脊柱的联合 数据以了解DRG神经元亚型可能如何与背角神经元的亚群相连。我们 将全面描述构成丘脑脊束的人类神经元。最后，在第三个目标中，我们将 利用人类背根神经节和脊髓上的核糖体图谱来获得对这些基因翻译事件的新见解 纸巾。长读测序将用于映射一组mRNAs中的RNA修饰和剪接 对疼痛来说很关键。我们的工作将创建一份全面的转录组、表转录组和 组成疼痛通路中第一个突触的神经元的翻译组，包括在慢性疼痛条件下。

There are clear differences between mouse and human nociceptors that have profound implications for the development of pain therapeutics that target the peripheral nervous system. Our spatial sequencing experiments reveal a distinct set of cell types in the human dorsal root ganglion (DRG). In this research project, we will compare DRG tissues from organ donors and surgical patients with chronic pain. We will include a range of ages across the adult life-span in both men and women in an ethnically diverse population. In addition to the DRG, we will also examine the spinal cord. We have already conducted SPLiT-seq and spatial transcriptomic analysis of dorsal and ventral horn from a select number of organ donors demonstrating the viability of both techniques. Our data shows that like the DRG, there are clear species differences in the spinal cord highlighting the importance of this discovery work. In our first aim we will use spatial sequencing and SPLiT-seq technologies to precisely define the cell types of the human DRG. We will use organ donor and pain patient DRGs to define how cellular transcriptomes change with pain, and if the changes covary with sex and age. SN ATAC-seq will identify transcriptional states and regulatory elements in human DRG. In our second aim we will use the same technologies to map the cell types of the human dorsal horn. We will strive to use the combined DRG and spinal cord data to understand how DRG neuron subtypes are likely to connect to subsets of dorsal horn neurons. We will fully characterize the human neurons that comprise the spino-thalamic tract. Finally, in our third aim we will use ribosome profiling on human DRG and spinal cord to gain new insight into translational events in these tissues. Long-read sequencing will be used to map RNA modifications and splicing in a set of mRNAs that are critical for pain. Our work will create a comprehensive atlas of the transcriptome, epi-transcriptome, and translatome of neurons that comprise the first synapses in the pain pathway, including in chronic pain conditions.