Label-free detection of prostaglandin transporter (SLCO2A1) function and inhibition: insights by wound healing and TRACT assays.
| Title: | Label-free detection of prostaglandin transporter (SLCO2A1) function and inhibition: insights by wound healing and TRACT assays. |
|---|---|
| Authors: | Mocking TAM; Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, Netherlands.; van Oostveen WM; Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, Netherlands.; Department of Rheumatology, Leiden University Medical Center, Leiden University, Leiden, Netherlands.; Immunology and Respiratory Diseases, Boehringer-Ingelheim, Ridgefield, CT, United States.; Oncode Institute, Leiden, Netherlands.; van Veldhoven JPD; Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, Netherlands.; Department of Rheumatology, Leiden University Medical Center, Leiden University, Leiden, Netherlands.; Immunology and Respiratory Diseases, Boehringer-Ingelheim, Ridgefield, CT, United States.; Oncode Institute, Leiden, Netherlands.; Minnee H; Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, Netherlands.; Fehres CM; Department of Rheumatology, Leiden University Medical Center, Leiden University, Leiden, Netherlands.; Whitehurst CE; Immunology and Respiratory Diseases, Boehringer-Ingelheim, Ridgefield, CT, United States.; IJzerman AP; Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, Netherlands.; Heitman LH; Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, Netherlands.; Oncode Institute, Leiden, Netherlands. |
| Source: | Frontiers in pharmacology [Front Pharmacol] 2024 May 09; Vol. 15, pp. 1372109. Date of Electronic Publication: 2024 May 09 (Print Publication: 2024). |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: Frontiers Media] Country of Publication: Switzerland NLM ID: 101548923 Publication Model: eCollection Cited Medium: Print ISSN: 1663-9812 (Print) Linking ISSN: 16639812 NLM ISO Abbreviation: Front Pharmacol Subsets: PubMed not MEDLINE |
| Imprint Name(s): | Original Publication: [Lausanne : Frontiers Media] |
| Abstract: | The prostaglandin transporter (PGT, SLCO2A1) mediates transport of prostanoids (a.o. prostaglandin E2 (PGE2)) into cells and thereby promotes their degradation. Overexpression of PGT leads to low extracellular PGE2 levels and has been linked to impaired wound healing of diabetic foot ulcers. Inhibition of PGT could thus be beneficial, however, no PGT inhibitors are currently on the market and drug discovery efforts are hampered by lack of high-through screening assays for this transporter. Here we report on a label-free impedance-based assay for PGT that measures transport activity through receptor activation (TRACT) utilizing prostaglandin E2 receptor subtype EP3 and EP4 that are activated by PGE2. We found that induction of PGT expression on HEK293-JumpIn-SLCO2A1 cells that also express EP3 and EP4 leads to an over 10-fold reduction in agonistic potency of PGE2. PGE2 potency could be recovered upon inhibition of PGT-mediated PGE2 uptake with PGT inhibitors olmesartan and T26A, the potency of which could be established as well. Moreover, the TRACT assay enabled the assessment of transport function of PGT natural variants. Lastly, HUVEC cells endogenously expressing prostanoid receptors and PGT were exploited to study wound healing properties of PGE2 and T26A in real-time using a novel impedance-based scratch-induced wound healing assay. These novel impedance-based assays will advance PGT drug discovery efforts and pave the way for the development of PGT-based therapies.; (Copyright © 2024 Mocking, van Oostveen, van Veldhoven, Minnee, Fehres, Whitehurst, IJzerman and Heitman.) |
| Competing Interests: | The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. |
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| Contributed Indexing: | Keywords: PGE2; SLCO2A1; genetic variants; label-free; prostaglandin transporter; prostanoid receptors; wound healing; xCELLigence |
| Entry Date(s): | Date Created: 20240524 Latest Revision: 20240525 |
| Update Code: | 20260130 |
| PubMed Central ID: | PMC11111933 |
| DOI: | 10.3389/fphar.2024.1372109 |
| PMID: | 38783936 |
| Database: | MEDLINE |
Journal Article