α-Synuclein expression in response to bacterial ligands and metabolites in gut enteroendocrine cells: an in vitro proof of concept study

Michael J Hurley; Elisa Menozzi; Sofia Koletsi; Rachel Bates; Matthew E Gegg; Kai-Yin Chau; Hervé M Blottière; Jane Macnaughtan; Anthony H V Schapira

doi:10.1093/braincomms/fcad285

α-Synuclein expression in response to bacterial ligands and metabolites in gut enteroendocrine cells: an in vitro proof of concept study

Brain Commun. 2023 Oct 24;5(6):fcad285. doi: 10.1093/braincomms/fcad285. eCollection 2023.

Authors

Michael J Hurley^{1

2}, Elisa Menozzi^{1

2}, Sofia Koletsi^{1

2}, Rachel Bates^{1

2}, Matthew E Gegg^{1

2}, Kai-Yin Chau^{1

2}, Hervé M Blottière^{2

3}, Jane Macnaughtan^{1

2

4}, Anthony H V Schapira^{1

2}

Affiliations

¹ Department of Clinical and Movement Neurosciences, UCL Queen Square Institute of Neurology, London NW3 2PF, UK.
² Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA.
³ Université Paris-Saclay, INRAE, MetaGenoPolis, Jouy en Josas, & Nantes Université, INRAE, UMR 1280 PhAN, Nantes 44093, France.
⁴ Institute for Liver and Digestive Health, University College London, London NW3 2PF, UK.

Abstract

Caudo-rostral migration of pathological forms of α-synuclein from the gut to the brain is proposed as an early feature in Parkinson's disease pathogenesis, but the underlying mechanisms remain unknown. Intestinal epithelial enteroendocrine cells sense and respond to numerous luminal signals, including bacterial factors, and transmit this information to the brain via the enteric nervous system and vagus nerve. There is evidence that gut bacteria composition and their metabolites change in Parkinson's disease patients, and these alterations can trigger α-synuclein pathology in animal models of the disorder. Here, we investigated the effect of toll-like receptor and free fatty acid receptor agonists on the intracellular level of α-synuclein and its release using mouse secretin tumour cell line 1 enteroendocrine cells. Secretin tumour cell line 1 enteroendocrine cells were treated for 24 or 48 h with toll-like receptor agonists (toll-like receptor 4 selective lipopolysaccharide; toll-like receptor 2 selective Pam3CysSerLys4) and the free fatty acid receptor 2/3 agonists butyrate, propionate and acetate. The effect of selective receptor antagonists on the agonists' effects after 24 hours was also investigated. The level of α-synuclein protein was measured in cell lysates and cell culture media by western blot and enzyme-linked immunosorbent assay. The level of α-synuclein and tumour necrosis factor messenger RNA was measured by quantitative reverse transcription real-time polymerase chain reaction. Stimulation of secretin tumour cell line 1 enteroendocrine cells for 24 and 48 hours with toll-like receptor and free fatty acid receptor agonists significantly increased the amount of intracellular α-synuclein and the release of α-synuclein from the cells into the culture medium. Both effects were significantly reduced by antagonists selective for each receptor. Toll-like receptor and free fatty acid receptor agonists also significantly increased tumour necrosis factor transcription, and this was effectively inhibited by corresponding antagonists. Elevated intracellular α-synuclein increases the likelihood of aggregation and conversion to toxic forms. Factors derived from bacteria induce α-synuclein accumulation in secretin tumour cell line 1 enteroendocrine cells. Here, we provide support for a mechanism by which exposure of enteroendocrine cells to specific bacterial factors found in Parkinson's disease gut dysbiosis might facilitate accumulation of α-synuclein pathology in the gut.

Keywords: enteroendocrine cells; free fatty acid receptors; microbiome; short-chain fatty acids; toll-like receptors.