Neuroactive steroid effects on autophagy in a human embryonic kidney 293 (HEK) cell model.

Autor: Salvatore SV; Departments of Psychiatry, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Ilagan MXG; High-Throughput Screening Core, Center for Drug Discovery, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Shu H; Departments of Psychiatry, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Lambert PM; Departments of Psychiatry, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA.; Medical Scientist Training Program, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Benz A; Departments of Psychiatry, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Qian M; Developmental Biology, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Covey DF; Developmental Biology, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA.; Taylor Family Institute for Innovative Psychiatric Research, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Zorumski CF; Departments of Psychiatry, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA.; Taylor Family Institute for Innovative Psychiatric Research, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA., Mennerick S; Departments of Psychiatry, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA. menneris@wustl.edu.; Taylor Family Institute for Innovative Psychiatric Research, Washington University in St. Louis School of Medicine, 660 S. Euclid Ave., MSC 8134-0181-0G, St. Louis, MO, 63110, USA. menneris@wustl.edu.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Jan 10; Vol. 14 (1), pp. 1042. Date of Electronic Publication: 2024 Jan 10.
DOI: 10.1038/s41598-024-51582-x
Abstrakt: Neuropsychiatric and neurodegenerative disorders are correlated with cellular stress. Macroautophagy (autophagy) may represent an important protective pathway to maintain cellular homeostasis and functionality, as it targets cytoplasmic components to lysosomes for degradation and recycling. Given recent evidence that some novel psychiatric treatments, such as the neuroactive steroid (NAS) allopregnanolone (AlloP, brexanolone), may induce autophagy, we stably transfected human embryonic kidney 293 (HEK) cells with a ratiometric fluorescent probe to assay NAS effects on autophagy. We hypothesized that NAS may modulate autophagy in part by the ability of uncharged NAS to readily permeate membranes. Microscopy revealed a weak effect of AlloP on autophagic flux compared with the positive control treatment of Torin1. In high-throughput microplate experiments, we found that autophagy induction was more robust in early passages of HEK cells. Despite limiting studies to early passages for maximum sensitivity, a range of NAS structures failed to reliably induce autophagy or interact with Torin1 or starvation effects. To probe NAS in a system where AlloP effects have been shown previously, we surveyed astrocytes and again saw minimal autophagy induction by AlloP. Combined with other published results, our results suggest that NAS may modulate autophagy in a cell-specific or context-specific manner. Although there is merit to cell lines as a screening tool, future studies may require assaying NAS in cells from brain regions involved in neuropsychiatric disorders.
(© 2024. The Author(s).)
Databáze: MEDLINE
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