TMT-based quantitative proteomic assessment of Vicia sativa induced neurotoxicity by β-cyano-L-alanine and γ-glutamyl-β-cyano-L-alanine in SH-SY5Y cells

Riley, Samuel; Nguyen, Vy; Bhattacharjee, Rudrarup; Ng, Pei Qin; Ritchie, Tarin; Fisk, Ian; Gecz, Jozef; Kumar, Raman; Searle, Iain R

Abstract

β-Cyano-L-alanine (BCA) and γ-glutamyl-β-cyano-L-alanine (GBCA) are the primary antinutritional compounds present within the seeds of the high protein and drought tolerant orphan legume Vicia sativa. Evidence of neurotoxicity is limited to symptom analysis from animal feed trials whilst the molecular mechanisms underlying suspected neurotoxicity in monogastric animals are largely unknown. In this study, we first optimised an in vitro cell-based assay for rapid testing of BCA and GBCA toxicity in the retinoic acid differentiated SH-SY5Y human neuroblastoma cells. Using this system, we then performed proteomics analyses to determine dysregulated expression of proteins in BCA or GBCA treated of differentiated SH-SY5Y cells. Our findings indicate that BCA affects expression of proteins involved in DNA damage and translation whilst GBCA treatment causes dysregulation of those involved in mitosis and cell cycle. Following BCA treatment, we identified changes in many proteins previously suggested to have close association with neurodegenerative diseases including amyotrophic lateral sclerosis and Alzheimer’s disease as well as cancers. Following GBCA treatment, dysregulation of proteins involved in apoptosis pathways was observed. Finally, the lack of common dysregulated proteins and pathways in BCA or GBCA treated cells indicated that they most likely cause neurotoxicity via distinct mechanisms.

Citation

Riley, S., Nguyen, V., Bhattacharjee, R., Ng, P. Q., Ritchie, T., Fisk, I., Gecz, J., Kumar, R., & Searle, I. R. (2025). TMT-based quantitative proteomic assessment of Vicia sativa induced neurotoxicity by β-cyano-L-alanine and γ-glutamyl-β-cyano-L-alanine in SH-SY5Y cells

• Working Paper Type: Preprint
• Publication Date: Apr 9, 2025
• Deposit Date: Apr 17, 2025
• Publicly Available Date: Apr 22, 2025
• DOI: https://doi.org/10.1101/2025.04.05.647336
• Public URL: https://nottingham-repository.worktribe.com/output/47828530
• Publisher URL: https://www.biorxiv.org/content/10.1101/2025.04.05.647336v1