eprintid: 10175357 rev_number: 7 eprint_status: archive userid: 699 dir: disk0/10/17/53/57 datestamp: 2023-08-17 13:41:24 lastmod: 2023-08-17 13:41:24 status_changed: 2023-08-17 13:41:24 type: article metadata_visibility: show sword_depositor: 699 creators_name: Wehrli, L creators_name: Galdadas, I creators_name: Voirol, L creators_name: Smieško, M creators_name: Cambet, Y creators_name: Jaquet, V creators_name: Guerrier, S creators_name: Gervasio, FL creators_name: Nef, S creators_name: Rahban, R title: The action of physiological and synthetic steroids on the calcium channel CatSper in human sperm ispublished: pub divisions: UCL divisions: B04 divisions: C06 divisions: F56 keywords: CatSper, steroids, human sperm, high-throughput screening, calcium signaling, pharmacophore modelling, acrosome reaction note: © 2023 Wehrli, Galdadas, Voirol, Smieško, Cambet, Jaquet, Guerrier, Gervasio, Nef and Rahban. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/). abstract: The sperm-specific channel CatSper (cation channel of sperm) controls the intracellular Ca2+ concentration ([Ca2+]i) and plays an essential role in sperm function. It is mainly activated by the steroid progesterone (P4) but is also promiscuously activated by a wide range of synthetic and physiological compounds. These compounds include diverse steroids whose action on the channel is so far still controversial. To investigate the effect of these compounds on CatSper and sperm function, we developed a high-throughput screening (HTS) assay to measure changes in [Ca2+]i in human sperm and screened 1,280 approved and off-patent drugs including 90 steroids from the Prestwick chemical library. More than half of the steroids tested (53%) induced an increase in [Ca2+]i and reduced the P4-induced Ca2+ influx in human sperm in a dose-dependent manner. Ten of the most potent steroids (activating and P4-inhibiting) were selected for a detailed analysis of their action on CatSper and their ability to act on sperm acrosome reaction (AR) and penetration in viscous media. We found that these steroids show an inhibitory effect on P4 but not on prostaglandin E1-induced CatSper activation, suggesting that they compete for the same binding site as P4. Pregnenolone, dydrogesterone, epiandrosterone, nandrolone, and dehydroepiandrosterone acetate (DHEA) were found to activate CatSper at physiologically relevant concentrations within the nanomolar range. Like P4, most tested steroids did not significantly affect the AR while stanozolol and estropipate slightly increased sperm penetration into viscous medium. Furthermore, using a hybrid approach integrating pharmacophore analysis and statistical modelling, we were able to screen in silico for steroids that can activate the channel and define the physicochemical and structural properties required for a steroid to exhibit agonist activity against CatSper. Overall, our results indicate that not only physiological but also synthetic steroids can modulate the activity of CatSper with varying potency and if bound to CatSper prior to P4, could impair the timely CatSper activation necessary for proper fertilization to occur. date: 2023-07-20 date_type: published publisher: Frontiers Media SA official_url: https://doi.org/10.3389/fcell.2023.1221578 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 2044712 doi: 10.3389/fcell.2023.1221578 lyricists_name: Gervasio, Francesco Luigi lyricists_id: FLGER36 actors_name: Flynn, Bernadette actors_id: BFFLY94 actors_role: owner full_text_status: public publication: Frontiers in Cell and Developmental Biology volume: 11 article_number: 1221578 citation: Wehrli, L; Galdadas, I; Voirol, L; Smieško, M; Cambet, Y; Jaquet, V; Guerrier, S; ... Rahban, R; + view all <#> Wehrli, L; Galdadas, I; Voirol, L; Smieško, M; Cambet, Y; Jaquet, V; Guerrier, S; Gervasio, FL; Nef, S; Rahban, R; - view fewer <#> (2023) The action of physiological and synthetic steroids on the calcium channel CatSper in human sperm. Frontiers in Cell and Developmental Biology , 11 , Article 1221578. 10.3389/fcell.2023.1221578 <https://doi.org/10.3389/fcell.2023.1221578>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10175357/1/fcell-11-1221578.pdf