Human-Induced Pluripotent Stem Cells (iPSCs) Differentiation Into Sensory Neurons for Screening Analgesic Compounds

• Grant number: 25/02959-0
• Support Opportunities: Regular Research Grants
• Start date: July 01, 2025
• End date: August 31, 2026
• Field of knowledge: Biological Sciences - Pharmacology - General Pharmacology
• Mobility Program: SPRINT - Projetos de pesquisa - Mobilidade
• Principal Investigator: Vanessa Olzon Zambelli
• Grantee: Vanessa Olzon Zambelli
• Principal researcher abroad: Juliana Milani Scorisa Salgado
• Institution abroad: Yale School of Medicine (YSM), United States
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated researchers: Michelle Cristiane Búfalo
• Associated research grant: 20/13139-0 - Centre of Excellence in New Target Discovery, AP.PCPE

Abstract

Human-induced pluripotent stem cells (iPSC) are promising for modeling diseases in individual human genetic backgrounds thus, for developing precision medicine. Also, their pluripotency allows them to be differentiated into diverse cell types. Previously, we showed that the neuroblastoma SH-SY5Y cell line is successfully differentiated into sensory neuron-like cells, expressing critical proteins involved in the nociceptive transmission, such as TRPV1, Nav1.7, Nav1.8, and Nav1.9. Importantly, challenging these cells with pro-inflammatory compounds is sufficient to up-regulate c-Fos expression, a marker for neuronal activity and pro-nociceptive gene expressions, as well as induce substance P release. Of interest, the incubation with morphine decreases substance P release, showing that these neurons are responsive to analgesics. Using this cell model, the Center of Excellence in New Target Discovery (CENTD) team found around five candidates with analgesic properties and one new molecular target that may regulate pain. However, there are limitations in differentiating SH-SY5Y in sensory neurons since it is derived from malignant tumors and, even after differentiation, the physiology may be compromised. Of note, the iPSC-derived neurons have the most physiologically relevant biology for recapitulating human neuron physiology as seen in vivo. This proposal represents a joint effort of two laboratories, capitalizing on their complementary expertise in pain models (CENTD/Butantan Institute) and pluripotent stem cells (iPSCs) to develop custom-tailored protocols designed for maximum efficiency (NeuroCore/ Yale University). The proposed exchange activities will include joint laboratory sessions, where both teams will work together to differentiate iPSCs into sensory neurons, thereby enabling a cutting-edge model for validating analgesic compounds and molecular targets for pain control. We expect to significantly contribute to the pain field and CENTD's mission, which is to identify molecular targets involved in inflammatory/painful diseases, using poisons and animal secretions as tools for developing new drugs. This collaboration will synergize our efforts in identifying effective analgesics, a critical area of study in chronic pain management. Together, we form a multidisciplinary team with behavioral pain experts, neuroscientists, and specialists in iPSC to provide translational applications for painful degenerative diseases. (AU)