Development of recombinant PLC-zeta protein as a therapeutic intervention for the clinical treatment of Oocyte Activation Failure

Saleh, Alaaeldin, Thanassoulas, Angelos, Aliyev, Elnur, Swann, Karl ORCID: https://orcid.org/0000-0002-4355-1449, Naija, Azza, Yalcin, Huseyin C., Lai, F. Anthony ORCID: https://orcid.org/0000-0003-2852-8547 and Nomikos, Michail 2024. Development of recombinant PLC-zeta protein as a therapeutic intervention for the clinical treatment of Oocyte Activation Failure. Biomedicines 12 (6) , 1183. 10.3390/biomedicines12061183

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Abstract

The sperm-specific phospholipase C zeta (PLCζ) protein is widely considered as the predominant physiological stimulus for initiating the Ca2+ release responsible for oocyte activation during mammalian fertilization. The increasing number of genetic and clinical reports that directly link PLCζ defects and/or deficiencies with oocyte activation failure (OAF) necessitates the use of a powerful therapeutic intervention to overcome such cases of male factor infertility. Currently, in vitro fertilization (IVF) clinics treat OAF cases after intracytoplasmic sperm injection (ICSI) with Ca2+ ionophores. Despite their successful use, such chemical agents are unable to trigger the physiological pattern of Ca2+ oscillations. Moreover, the safety of these ionophores is not yet fully established. We have previously demonstrated that recombinant PLCζ protein can be successfully used to rescue failed oocyte activation, resulting in efficient blastocyst formation. Herein, we produced a maltose binding protein (MBP)-tagged recombinant human PLCζ protein capable of inducing Ca2+ oscillations in mouse oocytes similar to those observed at fertilization. Circular dichroism (CD) experiments revealed a stable, well-folded protein with a high helical content. Moreover, the recombinant protein could retain its enzymatic properties for at least up to 90 days after storage at −80 °C. Finally, a chick embryo model was employed and revealed that exposure of fertilized chicken eggs to MBP-PLCζ did not alter the embryonic viability when compared to the control, giving a first indication of its safety. Our data support the potential use of the MBP-PLCζ recombinant protein as an effective therapeutic tool but further studies are required prior to its use in a clinical setting.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Biosciences
Additional Information:	License information from Publisher: LICENSE 1: URL: https://creativecommons.org/licenses/by/4.0/, Type: open-access
Publisher:	MDPI
Date of First Compliant Deposit:	10 June 2024
Date of Acceptance:	1 May 2024
Last Modified:	10 Jun 2024 09:00
URI:	https://orca.cardiff.ac.uk/id/eprint/169637

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