Abstract
Background
Growing evidence implicates early metabolic dysfunctions in retinal ganglion cells (RGCs) as a contributor to both high- and normal-tension glaucoma, yet no approved therapy directly protects RGCs to preserve vision. We aimed at identifying a safe, druggable neuroprotective strategy that restores RGC metabolic homeostasis for glaucoma therapy.
Methods
Using a live-cell mitochondrial screen in human embryonic stem cell-derived retinal ganglion cells (H7; female donor), we identified the clinically tested 5-HT1A antagonist WAY-100635 (WAY) as a neuroprotective agent. Mechanisms are probed by pharmacologic competition with agonist 8-OH-DPAT, cAMP assays, and PGC-1α dependent mitochondrial-biogenesis tests. RGC metabolism and survival are assessed by Seahorse and apoptosis assays. In vivo efficacy is evaluated in acute optic-nerve crush (ONC) and microbead-induced ocular-hypertension glaucoma models using histology, brain MRI, visual-acuity, contrast sensitivity testing, and flash VEPs to quantify cortical responses in wild-type C57BL/6 J male mice. Statistics used two-tailed Student’s t-tests or ANOVA, as appropriate.
Results
Here we show that WAY elicits a reversible cAMP surge that drives PGC-1α dependent mitochondrial biogenesis and reduces apoptosis in hRGCs. In glaucoma-associated OPTNE50K hRGCs, it restores mitochondrial fitness, attenuates excitotoxicity, and shifts metabolism toward aerobic glycolysis, while in progenitors, WAY enhances cristae maturation, oxidative phosphorylation, accelerating RGC specification. Systemic dosing in ONC mice preserves RGC somata, retinal function (PhNR), and optic-pathway integrity. WAY-treated glaucoma mice show preserved visual acuity and fVEP propagation to cortex, halting glaucoma progression.
Conclusions
A clinically tested 5-HT1A antagonist WAY restores RGC metabolic homeostasis and preserves visual-pathway function across acute and chronic injury models, without detected systemic toxicity, supporting development of a neuroprotective candidate for glaucoma and potentially for other mitochondrial optic neuropathies.
Plain Language Summary
Glaucoma slowly damages the nerve cells called retinal ganglion cells (RGCs) that carry signals from the eye to the brain. Current treatments mainly lower eye pressure but even when treated, many patients continue to lose vision. We screened for various possible compounds on human RGCs and discovered a drug already tested in people for another reason keeps RGCs alive during optic nerve injury and maintains the ability for visual signals to move from the eye to the brain, including in conditions where glaucoma develops. These results suggest this treatment could be used alongside pressure-lowering treatments to preserve vision. Further testing is needed to check this would be suitable for people with glaucoma.
Data availability
The resources used in this study have been detailed in the method and Supplemental table 1, and source data are available in Supplementary Data 2. Requests for further information and resources should be directed to and will be fulfilled by the lead contact, Arupratan Das (arupdas@iu.edu). All uncropped images for western blots are presented in the Supplementary Fig. 6 and flow cytometry gating steps are presented in the Supplementary Fig. 7.
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Acknowledgements
We thank the Hypoxia Core Facility at the Indiana University Cooperative Center for Excellence in Hematology (U54 DK106846) for Seahorse support, and Dr. Donald J. Zack for insightful feedback. Transmission-electron microscopy was performed with Derrick Gray, Dr. Quyen Q. Hoang, and Yangshin Park at the Indiana University School of Medicine (IUSM) Electron Microscopy Core; MRI sample preparation and imaging was assisted by Dr. Yu-Chien Wu and Erin E. Jarvis at the IU in vivo imaging core. We are thankful to Andi R. Masters and Christine M. Bach at the Clinical Pharmacology Analytical Core (CPAC, IUSM) for mass-spectrometry analyses, and to Drew M. Brown (Histology Core, IUSM) for tissue sectioning and H&E staining. We acknowledge Drs. Neha Mahajan, Qianyi Luo, and Ashay D. Bhatwadekar for OKR and ERG recordings; Drs. Amir R. Hajrasouliha and Sunland Gong for PhNR measurements; and Drs. Padmanabhan Pattabiraman and Avinash Soundararajan for IOP monitoring and anesthesia. Guidance in cryosectioning and staining was provided by Drs. Yoshikazu Imanishi, Sanae Imanishi, and Shimpei Takita; Dr. Weiming Mao for the access to his light-microscope; and Dr. Timothy W. Corson, Dr. Anbukkarasi Muniyandi, and Kamakshi Sishtla assisted with cell cycle analysis, enucleation and retinal dissection procedures. We thank Dr. Erica Cai (Indiana Biosciences Research Institute) for antisense-oligonucleotide (ASO) design, and Dr. Alyssa Coyne (Johns Hopkins University) for guidance on gymnotic ASO delivery. Drs. Thomas V. Johnson III and Erika A. Aguzzi (Johns Hopkins University) trained us optic-nerve crush and retinal flat-mount techniques; Dr. Harry A. Quigley and Elizabeth Kimball (Johns Hopkins University) assisted with the microbead glaucoma model. We thank Dr. Larry Benowitz and Dr. Yuqin Yin (Harvard Medical School) for providing the GAP-43 antibody, immunohistochemistry and image quantification protocols, and for reviewing GAP-43 positive optic nerve images. Funding was provided by the NIH (R00 EY028223 to A.D.; UL1 TR002529 to S.D. and A.D., R01 NS125020 to N.W.); Research to Prevent Blindness Challenge Grant to IU Ophthalmology, Indiana CTSI, The Glaucoma Foundation, Showalter Research Trust, Indiana University School of Medicine, and the BrightFocus Foundation (RReSTORe collaborative grant) to A. D. K.A. was supported in part by the Paul and Carole Stark Fellowship and a Sigma Xi Grant-in-Aid of Research. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
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A.D. conceived the study and supervised all aspects of the project. A.D., S.D., M.S., and K.A. designed the experiments, carried out the research, and analyzed the data. J.M. designed and executed the small-molecule screen. N.W. and J.C. processed, analyzed, and interpreted the MRI data.
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The authors declare no competing interests. A.D. is an inventor on a patent application (PCT/US2024/044204) currently under review by the U.S. Patent and Trademark Office, filed through Indiana University School of Medicine.
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Communications Medicine thanks Makoto Ishikawa, Richard Libby and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. A peer review file is available.
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Dutta, S., Surma, M.L., Chen, J. et al. The 5-HT1A receptor antagonist WAY-100635 maleate promotes retinal ganglion cell differentiation and protects the retino-visual circuits. Commun Med (2026). https://doi.org/10.1038/s43856-026-01528-3
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DOI: https://doi.org/10.1038/s43856-026-01528-3
