PROJECT SUMMARY/ABSTRACT Species-specific differences and the complexity of cellular interactions involved in human pain perception have created challenges relying on conventional preclinical rodent models or simple human pluripotent stem cell (hPSC)-derived nociceptor models for novel drug development. Pain signaling pathways comprise of multiple neuronal, glial, and immune cell types within the peripheral (PNS) and central nervous systems (CNS), but the contributions of these cells in pain sensation is still under investigation. Moreover, these cells and the circuits they form exhibit region-specific characteristics, with unique positional identities along the body axis, and require a three-dimensional (3D) microenvironment with component parts—including end-organ tissues— to establish appropriate connectivity, plasticity, excitability, and functionality. Recognizing these challenges, the goal of this project is to bridge the gap by developing advanced stem cell tools to facilitate the creation of comprehensive and region-specific models for studying afferent pain circuitry. Building upon established technologies for region-specific spinal cord differentiation within my lab, this project will pursue three main objectives. Objective 1 involves developing scalable and systematic methods to generate region-specific sensory neurons and glial subtypes. Objective 2 focuses on creating and characterizing region-specific somatosensory organoids that faithfully mimic the in vivo microenvironment. Objective 3 aims to integrate hPSC-derived pain circuits into two tissue-engineered models for women's health—breast cancer metastasis and endometriosis— to determine how region- and subtype- specificity impacts innervation in the context of health and disease. Altogether, we will produce a suite of region-specific cells, circuits, and tissues that will deepen our understanding of the unique pain networks along the body axis and expedite the development of model systems for high-throughput screening of targeted analgesics. Furthermore, as a new investigator with multidisciplinary interests, this New Innovator Award will advance technologies in our lab that can be applied beyond pain research to a broad range of applications in neurological diseases, tissue engineering, and regenerative medicine.