Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
  • Clinical Research and Public Health
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Video Abstracts
  • Reviews
    • View all reviews ...
    • Complement Biology and Therapeutics (May 2025)
    • Evolving insights into MASLD and MASH pathogenesis and treatment (Apr 2025)
    • Microbiome in Health and Disease (Feb 2025)
    • Substance Use Disorders (Oct 2024)
    • Clonal Hematopoiesis (Oct 2024)
    • Sex Differences in Medicine (Sep 2024)
    • Vascular Malformations (Apr 2024)
    • View all review series ...
  • Viewpoint
  • Collections
    • In-Press Preview
    • Clinical Research and Public Health
    • Research Letters
    • Letters to the Editor
    • Editorials
    • Commentaries
    • Editor's notes
    • Reviews
    • Viewpoints
    • 100th anniversary
    • Top read articles

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Video Abstracts
  • In-Press Preview
  • Clinical Research and Public Health
  • Research Letters
  • Letters to the Editor
  • Editorials
  • Commentaries
  • Editor's notes
  • Reviews
  • Viewpoints
  • 100th anniversary
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
GPR101 mediates the pro-resolving actions of RvD5n-3 DPA in arthritis and infections
Magdalena B. Flak, … , Francesco Palmas, Jesmond Dalli
Magdalena B. Flak, … , Francesco Palmas, Jesmond Dalli
Published December 3, 2019
Citation Information: J Clin Invest. 2020;130(1):359-373. https://doi.org/10.1172/JCI131609.
View: Text | PDF
Research Article Infectious disease Inflammation

GPR101 mediates the pro-resolving actions of RvD5n-3 DPA in arthritis and infections

  • Text
  • PDF
Abstract

N-3 docosapentaenoic acid–derived resolvin D5 (RvD5n-3 DPA) is diurnally regulated in peripheral blood and exerts tissue-protective actions during inflammatory arthritis. Here, using an orphan GPCR screening approach coupled with functional readouts, we investigated the receptor(s) involved in mediating the leukocyte-directed actions of RvD5n-3 DPA and identified GPR101 as the top candidate. RvD5n-3 DPA bound to GPR101 with high selectivity and stereospecificity, as demonstrated by a calculated KD of approximately 6.9 nM. In macrophages, GPR101 knockdown limited the ability of RvD5n-3 DPA to upregulate cyclic adenosine monophosphate, phagocytosis of bacteria, and efferocytosis. Inhibition of this receptor in mouse and human leukocytes abrogated the pro-resolving actions of RvD5n-3 DPA, including the regulation of bacterial phagocytosis in neutrophils. Knockdown of the receptor in vivo reversed the protective actions of RvD5n-3 DPA in limiting joint and gut inflammation during inflammatory arthritis. Administration of RvD5n-3 DPA during E. coli–initiated inflammation regulated neutrophil trafficking to the site of inflammation, increased bacterial phagocytosis by neutrophils and macrophages, and accelerated the resolution of infectious inflammation. These in vivo protective actions of RvD5n-3 DPA were limited when Gpr101 was knocked down. Together, our findings demonstrate a fundamental role for GPR101 in mediating the leukocyte-directed actions of RvD5n-3 DPA.

Authors

Magdalena B. Flak, Duco S. Koenis, Agua Sobrino, James Smith, Kimberly Pistorius, Francesco Palmas, Jesmond Dalli

×

Figure 3

RvD5n-3 DPA stereospecifically activates GPR101 and increases cAMP in human macrophages.

Options: View larger image (or click on image) Download as PowerPoint
RvD5n-3 DPA stereospecifically activates GPR101 and increases cAMP in hu...
(A) GPR101-expressing CHO cells were incubated with RvD5n-3 DPA or DA-RvD5n-3 DPA (10 nM), and impedance was measured for 30 minutes. Results are representative of 3 distinct experiments. (B) GPR101-expressing CHO cells were incubated with the indicated concentrations of the ligands described in A, and changes in impedance from baseline values were determined at t = 10 minutes. Results represent the mean ± SEM. n = 3 from 3 distinct experiments. *P < 0.05; 1-way ANOVA with a Holm-Sidak post hoc multiple comparisons test. (C) GPR101-overexpressing CHO cells were incubated with RvD5n-3 DPA or DHA-derived RvD5 (1 nM), and cell impedance was measured over a 30-minute period using the xCELLigence DP system. Results are shown as the mean ± SEM (n = 4 in 3 independent experiments). (D) CHO cells expressing GPR101 coupled with the β-arrestin luminescence reporter system were incubated with the indicated concentrations of RvD5n-3 DPA, DHA-derived RvD5, or vehicle (cell-plating reagent containing 0.01% ethanol), and receptor activation was measured as an increase in luminescence signal. Results are shown as the mean ± SEM (n = 3 in 2 independent experiments). (E) GPR101-expressing CHO cells were incubated with CTX (1 μg/mL, 2 hours), PTX (1 μg/mL, 16 hours), or vehicle and then with RvD5n-3 DPA (10 nM), and impedance was measured over a 30-minute period. Results are representative of 3 distinct experiments. (F) Human monocyte–derived macrophages were incubated with either an siRNA against GPR101 or a control sequence (CT siRNA; 72 hours at 37°C) and then with RvD5n-3 DPA (10 nM) or vehicle (Veh) (PBS containing 0.01% ethanol) for 2 minutes, and cAMP concentrations were assessed. Results represent the mean ± SEM (n = 4 donors). *P < 0.05; 1-way ANOVA with Holm-Sidak post hoc multiple comparisons test.

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts