Anti-Inflammatory Properties of Chemical Probes in Human Whole Blood: Focus on Prostaglandin E2 Production.

Filip Bergqvist,Per-Johan Jakobsson,Yvonne Sundström,Ingrid E Lundberg,Louise Berg,Michael Sundström,Iva Gunnarsson,Ming-Mei Shang

doi:10.3389/fphar.2020.00613

Filip Bergqvist, Per-Johan Jakobsson + Show 6 more

Open Access

https://doi.org/10.3389/fphar.2020.00613

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Abstract

We screened 57 chemical probes, high-quality tool compounds, and relevant clinically used drugs to investigate their effect on pro-inflammatory prostaglandin E2 (PGE2) production and interleukin-8 (IL-8) secretion in human whole blood. Freshly drawn blood from healthy volunteers and patients with systemic lupus erythematosus (SLE) or dermatomyositis was incubated with compounds at 0.1 or 1 µM and treated with lipopolysaccharide (LPS, 10 µg/ml) to induce a pro-inflammatory condition. Plasma was collected after 24 h for lipid profiling using liquid chromatography tandem mass spectrometry (LC-MS/MS) and IL-8 quantification using enzyme-linked immunosorbent assay (ELISA). Each compound was tested in at least four donors at one concentration based on prior knowledge of binding affinities and in vitro activity. Our screening suggested that PD0325901 (MEK-1/2 inhibitor), trametinib (MEK-1/2 inhibitor), and selumetinib (MEK-1 inhibitor) decreased while tofacitinib (JAK inhibitor) increased PGE2 production. These findings were validated by concentration–response experiment in two donors. Moreover, the tested MEK inhibitors decreased thromboxane B2 (TXB2) production and IL-8 secretion. We also investigated the lysophophatidylcholine (LPC) profile in plasma from treated whole blood as these lipids are potentially important mediators in inflammation, and we did not observe any changes in LPC profiles. Collectively, we deployed a semi-high throughput and robust methodology to investigate anti-inflammatory properties of new chemical probes.

Highlights

Inflammation is a highly controlled immune response to eliminate the cause of tissue injury or infection and to initiate tissue repair back to homeostasis via resolution (Nathan, 2002; Buckley et al, 2013)
We have tested the inhibitory effect on prostanoid production and IL-8 secretion in human whole blood for 57 high-quality inhibitors with known target specificities and in vitro potencies
We found that our positive control diclofenac for blocking prostanoid production decreased IL-8 secretion, which is explained by the fact that Prostaglandin E2 (PGE2) stimulates IL-8 production in cultured cells (Agro et al, 1996; Caristi et al, 2005; Aso et al, 2012; Venza et al, 2012)

Summary

Introduction

Inflammation is a highly controlled immune response to eliminate the cause of tissue injury or infection and to initiate tissue repair back to homeostasis via resolution (Nathan, 2002; Buckley et al, 2013). Inflammatory responses drive many autoimmune diseases (McInnes and Schett, 2011) and inflammation is a hallmark of cancer (Hanahan and Weinberg, 2011). Prostaglandin E2 (PGE2) is a potent lipid mediator of inflammation and immune responses, and PGE2 is a central mediator of pain, edema, and cartilage erosion typically observed in the joints of rheumatoid arthritis patients (Akaogi et al, 2012; Fattahi and Mirshafiey, 2012). PGE2 is synthesized via conversion of arachidonic acid by cyclooxygenases (COX-1 and COX-2) into unstable PGH2 that is further metabolized by the inducible terminal synthase microsomal prostaglandin E synthase-1 (mPGES-1) to generate PGE2. Selective inhibition of PGE2 production with small molecule inhibitors could be a desirable therapeutic strategy in inflammation and cancer (Bergqvist et al, 2020)

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