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

小鼠和人类的伤害感受器之间存在明显的差异，这些差异对神经系统的发育有着深远的影响。 开发针对周围神经系统的疼痛治疗方法。我们的空间测序实验 揭示了人类背根神经节（DRG）中一组独特的细胞类型。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手术患者的DRG组织。我们将包括一系列的年龄 在不同种族的人群中，男性和女性在成年后的整个生命周期中都有这种变化。除了DRG，我们 也会检查脊髓我们已经进行了SPLiT-seq和空间转录组学分析， 背角和腹角从一个选定的一些器官捐赠者证明了这两种技术的可行性。我们 数据显示，像背根神经节一样，脊髓中存在明显的物种差异， 这一发现工作。在我们的第一个目标中，我们将使用空间测序和SPLiT-seq技术来精确地 定义人类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来定义细胞如何 转录组随着疼痛而改变，并且如果改变与性别和年龄协变。SN ATAC-seq将识别 转录状态和调节元件。在我们的第二个目标中，我们将使用相同的 技术来绘制人类背角的细胞类型。我们将努力使用联合DRG和脊髓 脊髓数据，以了解DRG神经元亚型可能如何连接到背角神经元的子集。我们 将充分表征构成脊髓丘脑束的人类神经元。最后，在我们的第三个目标中， 使用核糖体分析人类DRG和脊髓，以获得新的见解，在这些翻译事件 组织中长读段测序将用于绘制一组mRNA中的RNA修饰和剪接， 对疼痛至关重要。我们的工作将创建一个全面的图谱的转录组，外转录组， 在疼痛通路中，包括在慢性疼痛病症中，神经元的翻译组包括第一突触。

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.