Abstract: Tens of millions of Americans suffer from chronic pain. Opioids represent the main tool for treating pain, but their use in chronic pain conditions suffers from a poor evidence base and the inherent risk of addiction. The current crisis of opioid-related deaths highlights the risk associated with widespread opioid use. The PRECISION Human Pain Network, part of the NIH HEAL Initiative, seeks to provide foundational data on the diversity among cell types that comprise the pain neuraxis. Here, we propose a U19 program entitled “Integrated Research Center for Human Pain Tissues” (INTERCEPT Pain). We will build on prior successes in our human tissue research program, and expand the scope of work to directly address the goals of the PRECISION Human Pain Network. Our program leverages the world-class genomics and computational assets of the McDonnell Genome Institute here at Washington University and substantial institutional strengths in neurobiology of pain, axon degeneration and regeneration, and genetics in a coordinated program to develop foundational new knowledge regarding the transcriptional and functional properties of tissues and cells involved in pain transduction, transmission and modulation in humans. We will provide a comprehensive atlas of cellular gene expression in human peripheral nerve using single nuclei RNAseq, spatial transcriptomics and multiplex proteomics, and leverage this to understand the distribution of genes involved in traumatic (neuromas) and idiopathic (hereditary) painful neuropathies. We will also provide expanded single-nucleus sequencing atlases of dorsal root ganglia, including DRG from donors without and with a recent history of pain. We will generate a spatial atlas of hDRG using imaging mass cytometry, and optimize computational approaches for integrated IMC and single cell transcriptomic analysis of hDRG. Finally, we will combine analysis of electrophysiological, transcriptional, and morphological data from hDRG and human spinal cord dorsal horn neurons. We will work with other centers to optimize protocols and data collection to allow integrated analysis across multiple centers, in collaboration with the U24 DCIC and HEAL data ecosystem. These goals will be accomplished through the coordinated activity of 3 scientific projects, led by international leaders in the fields of pain neurobiology, genetics, and mechanisms of axon degeneration and regeneration. The project titles are as follows: Project 1: Multi-omics peripheral nerve atlas enables fine-mapping of pain molecular phenotypes. Project 2: Characterization of the human dorsal root ganglia at the single cell level via integrated transcriptomics and spatial proteomics Project 3: Functional and genetic characterization of human DRG and spinal cord at single cell resolution The projects are supported by an experienced program leadership team and an administration core, a human tissue procurement and processing core, and a data core.

Project Narrative: Tens of millions of Americans suffer from chronic pain, and the epidemic of overdose deaths related to opioids makes it clear that safer treatments are needed. To help develop new, safer treatments, scientists must learn much more about the genes and cells that contribute to pain in humans. This application proposes a new center that will map the genes and cells associated with various types of pain, focusing on the peripheral nerves and spinal cord, to provide important new information to help with the development of new therapies.