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bioRxiv · 10.1101/2025.06.28.662125

TRPML3 regulates neuronal gene expression in an in vitro model of autophagy and may act as a genetic marker of familial neurodegenerative disorders

Adusumilli, S.·Mathe, M. M.·Shandilya, J.·Nayak, T. K.
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Abstract

Autophagy is a conserved pro-survival pathway for delivering misfolded proteins and damaged organelles to lysosomes for degradation and protein homeostasis. Anomaly in autophagy leads to aberrant protein aggregation in neuronal cells, which is a common etiology of neurodegenerative disorders. Endo-lysosomal cation channel TRPML3 (Transient Receptor Potential Mucolipin-3) has been shown to induce autophagy in cell line models. However, the mechanism of TRPML3 mediated autophagy induction and the underlying gene expression changes are not clearly understood. Here, by using Ca2+-and electrical-current measurements, RNA sequencing and RT PCR studies, we explored the cellular function of TRPML3 and the global transcriptomic profile in a cell-based serum starvation model of autophagy. We report that serum starvation leads to downregulation of neuronal developmental genes during autophagy induction. TRPML3 overexpression further amplifies the effect of starvation in downregulating neuronal gene expression. But, when nutrition is not a limiting condition, TRPML3 overexpression upregulated neuronal genes including those responsible for axon guidance, synaptogenesis, and dendritic arborization. TRPML3 mediated neuronal gene expression changes were, presumably, due to transcription factors (TF) TFEB, FOXO1 and neuron-specific TFs such as SOX2, and ETV5. To further validate the role of TRPML3 in neuronal gene regulation, we performed meta-analysis of publicly available RNAseq datasets on neurodegenerative disorders which provided insight into the heterogeneity in the molecular mechanisms of autophagy and corroborated the TFEB-mediated autophagy induction and neuronal gene expression in TRPML3 overexpression condition. Based on our results, we propose that TRPML3 may act as a potential genetic marker for familial neurodegenerative disorders.

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Adusumilli, S., Mathe, M. M., Shandilya, J., Nayak, T. K.. 2025-06-29. TRPML3 regulates neuronal gene expression in an in vitro model of autophagy and may act as a genetic marker of familial neurodegenerative disorders. https://doi.org/10.1101/2025.06.28.662125

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