A novel blood-based bioassay to monitor adiponectin signaling.

Autor: Lone AH; Department of Biology, York University, Toronto, ON, Canada., Tang J; Department of Biology, York University, Toronto, ON, Canada., Pignalosa A; Allysta Pharmaceuticals Inc., Bellevue, WA, USA., Hsu HH; Allysta Pharmaceuticals Inc., Bellevue, WA, USA., Abdul-Sater AA; School of Kinesiology and Health Science, York University, Toronto, ON, Canada. Electronic address: aasater@yorku.ca., Sweeney G; Department of Biology, York University, Toronto, ON, Canada. Electronic address: gsweeney@yorku.ca.
Jazyk: angličtina
Zdroj: International immunopharmacology [Int Immunopharmacol] 2024 May 10; Vol. 132, pp. 111890. Date of Electronic Publication: 2024 Mar 27.
DOI: 10.1016/j.intimp.2024.111890
Abstrakt: The diverse beneficial effects of adiponectin-receptor signaling, including its impact on the regulation of inflammatory processes in vivo, have resulted in development of adiponectin receptor agonists as a treatment for metabolic disorders. However, there are no established non-invasive bioassays for detection of adiponectin target engagement in humans or animal models. Here, we designed an assay using small amounts of blood to assess adiponectin action. Specifically, we tested effects of the small 10-amino acid peptide adiponectin receptor agonist, ALY688, in a sublethal LPS endotoxemia model in mice. LPS-induced pro-inflammatory cytokine levels in serum were significantly reduced in mice treated with ALY688, assessed via multiplex ELISA in flow cytometry. Furthermore, ALY688 alone significantly induced TGF-β release in serum 1 h after treatment and was elevated for up to 24 h. Additionally, using a flow-cytometry panel for detection of changes in circulating immune cell phenotypes, we observed a significant increase in absolute T cell counts in mice after ALY688 treatment. To assess changes in intracellular signaling effectors downstream of adiponectin, phospho-flow cytometry was conducted. There was a significant increase in phosphorylation of AMPK and p38-MAPK in mice after ALY688 treatment. We then used human donor immune cells (PBMCs) treated with ALY688 ex vivo and observed elevation of AMPK and p38-MAPK phosphorylation from baseline in response to ALY688. Together, these results indicate we can detect adiponectin action on immune cells in vivo by assessing adiponectin signaling pathway for AMPK and p38-MAPK, as well as pro-inflammatory cytokine levels. This new approach provides a blood-based bioassay for screening adiponectin action.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: These studies were supported in part via a research contract with Allysta Pharmaceuticals Inc. HHH and AP are employees of Allysta and GS and AAS consult for Allysta. Other authors confirm that no conflict of interest occurs.
(Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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