Phenotypic drug screen uncovers the metabolic GCH1/BH4 pathway as key regulator of EGFR/KRAS-mediated neuropathic pain and lung cancer Cronin, Shane J. F. Rao, Shuan Tejada Giráldez, Miguel Ángel Turnes, Bruna Lenfers Licht-Mayer, Simon Omura, Takao Brenneis, Christian Jacobs, Emily Barret, Lee Latremoliere, Alban Andrews, Nick A. Channon, Keith M. Latini, Alexandra Arvanites, Anthony C. Davidow, Lance S. Costigan, Michael Rubin, Lee L. Penninger, Josef M. Woolf, J. Woolf C.J.W. is supported by NIH R35NS105076 and an HMS Blavatnik Award. J.M.P. is supported by the Austrian Federal Ministry of Education, Science and Research, the Austrian Academy of Sciences, and the city of Vienna and grants from the Austrian Science Fund (FWF) Wittgenstein award (Z 271-B19), the T. von Zastrow foundation, a Canada 150 Research Chairs Program (F18-01336), and CIHR (FRN 168899). L.L.R. is supported by an HSCI Therapeutic Screening Center (TSC) grant (no. CF-0009-17-03). A. Latremoliere is supported by NIH grant R01NS112266. A. Latini is a CNPq fellow. S.R. is supported by the National Key R&D Program of China, 2021YFE0193400. Increased tetrahydrobiopterin (BH4) generated in injured sensory neurons contributes to increased pain sensitivity and its persistence. GTP cyclohydrolase 1 (GCH1) is the rate-limiting enzyme in the de novo BH4 synthetic pathway, and human single-nucleotide polymorphism studies, together with mouse genetic modeling, have demonstrated that decreased GCH1 leads to both reduced BH4 and pain. However, little is known about the regulation of Gch1 expression upon nerve injury and whether this could be modulated as an analgesic therapeutic intervention. We performed a phenotypic screen using about 1000 bioactive compounds, many of which are target-annotated FDA-approved drugs, for their effect on regulating Gch1 expression in rodent injured dorsal root ganglion neurons. From this approach, we uncovered relevant pathways that regulate Gch1 expression in sensory neurons. We report that EGFR/KRAS signaling triggers increased Gch1 expression and contributes to neuropathic pain; conversely, inhibiting EGFR suppressed GCH1 and BH4 and exerted analgesic effects, suggesting a molecular link between EGFR/KRAS and pain perception. We also show that GCH1/BH4 acts downstream of KRAS to drive lung cancer, identifying a potentially druggable pathway. Our screen shows that pharmacologic modulation of GCH1 expression and BH4 could be used to develop pharmacological treatments to alleviate pain and identified a critical role for EGFR-regulated GCH1/BH4 expression in neuropathic pain and cancer in rodents. 2025-01-27T08:06:05Z 2025-01-27T08:06:05Z 2022 journal article Published version: Cronin SJF. et al. Phenotypic drug screen uncovers the metabolic GCH1/BH4 pathway as key regulator of EGFR/KRAS-mediated neuropathic pain and lung cancer. Sci Transl Med. 2022 Aug 31;14(660):eabj1531. doi: 10.1126/scitranslmed.abj1531 https://hdl.handle.net/10481/100393 10.1126/scitranslmed.abj1531 eng http://creativecommons.org/licenses/by-nc-nd/4.0/ open access Attribution-NonCommercial-NoDerivatives 4.0 Internacional American Association for the Advancement of Science