Atlas-Guided Cell-Specific Transcriptomics Identifies Pathway-Level Remodeling of Human Hypothalamic CRH Neurons in Opioid Use Disorder

Abstract

Opioid use disorder (OUD) disrupts hypothalamic stress signaling, yet the molecular state of human paraventricular nucleus (PVN) corticotropin-releasing hormone (CRH) neurons remains poorly defined. Using single-nucleus RNA sequencing of 343,819 human hypothalamic nuclei, we applied complementary human HYPOMAP/MapMyCells and mouse PVN Atlas strategies that independently converged on the standardized Allen Brain Map subcluster Splat 410 843, providing cross-atlas validation of a rare CRH-enriched PVN population. Integrated GO, Reactome and KEGG analysis revealed OUD-associated enrichment of synaptic, junctional, cytoskeletal, receptor-signaling and ion transport programs, with relative enrichment of RNA processing, ER Golgi/vesicular, endolysosomal and glycan-related functions in controls. Targeted neuropeptide analysis further identified nominal increases in OXT, GHR, GAL and PRLR and decreases in VIP, AGRP, NPY, CRHBP and CALCR. Together, these findings define a reproducible human PVN CRH population and identify coordinated pathway and neuropeptide remodeling in OUD.