Effect of COVID-19 on Platelet Mitochondrial Bioenergetic, Antioxidants and Oxidative Stress in Infertile Men.

COVID-19 Clinical Trial

— COVInfertility  

Official title:

Effect of COVID-19 on Spermiogram, Platelet Mitochondrial Bioenergetic, Antioxidants and Oxidative Stress in Infertile Men

Clinical Trial Details — Status: Active, not recruiting

Administrative data

• NCT number: NCT05421234
• Other study ID #: COVID-19-INFERTILITY-SK01/2022
• Status: Active, not recruiting
• Phase: N/A
• Start date: April 1, 2022
• Est. completion date: March 30, 2023

Study information

• Verified date: January 2022
• Source: Comenius University
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

To verify the hypothesis that infertility and the effect of SARS-CoV-2 on infertility may damage platelet mitochondrial bioenergetics and endogenous coenzyme Q10 levels in infertile men.

Description:

Infertility is defined as the failure of the reproductive system to achieve pregnancy after 12 months of unprotected sex life. The pathobiochemical mechanisms of male fertility disorders include reduced sperm motility and quality, oxidative stress, reduced antioxidant capacity, mtDNA fragmentation, and sperm mitochondrial dysfunction.

Sperm contain a number of mitochondria that are spirally arranged around the middle part of the axomen. The main role of mitochondria in spermatozoa is to generate the energy needed for their motility (1, 2). Endogenous sources - coenzyme Q10 and carnitine - are key for energy production (ATP) in sperm mitochondria. Physiological functions of sperm require a minimal amount of reactive oxygen species (ROS), but uncontrolled ROS production contributes to reduced motility and sperm count, fragmentation of mtDNA (3).

In recent years, blood cells (platelets, lymphocytes and monocytes) have been used to diagnose mitochondrial disorders. Isolated peripheral blood platelets are an available source of mitochondria to assess mitochondrial health. Platelets receive energy mainly through glycolysis and oxidative phosphorylation. Platelet mitochondrial dysfunction has been demonstrated in patients with chronic kidney disease (4, 5), in patients with rheumatoid arthritis (6), in patients with acute COVID-19 (7). An O2k-respirometer (Oroboros, Austria) (8, 9) is used for respirometric analysis of platelet mitochondrial bioenergetics.

None information is available on the effect of infertility on platelet mitochondrial function, none on the effect of SARS-CoV-2 on platelet mitochondrial function in infertile patients, or the effect of vaccination on sperm function. Testicular damage and subsequent infertility due to SARS-CoV infection is expected. -2, directly via SARS-CoV-2 binding to ACE2 receptors or secondarily, in relation to the immunological and inflammatory response (10). SARS-CoV-2 virus induces excessive production of pro-inflammatory cytokines, mainly interleukin 6 (IL6), interleukin-1β (IL-1β) and tumor necrosis factor α (TNFα). Cytokines can impair sperm movement and reduce sperm count. High levels of pro-inflammatory cytokines have been found in infertile men (with oligozoospermia, asthenozoospermia, teratozoospermia) (11).

SARS-CoV-2 virus can manipulate mitochondrial function in patients with post-COVID-19 syndrome, which may persist for a long time (12). In previous our study the investigators found modulation of platelet mitochondrial respiration, reduction ATP production via oxidative phosphorylation, reduces endogenous coenzyme Q10 production, reprogramming of cellular metabolism patients after 4-7 weeks overcoming acute COVID-19, SARS-CoV-2 (7). In another studies the investigators confirmed platelet mitochondrial bioenergetic deficiency, reduced endogenous coenzyme Q10 production in patients with post-COVID-19 syndrome, 3-6 months after overcoming COVID-19 (13, 14, 15). Results of this study contribute to the understanding of the pathobiochemical mechanisms of infertility on subcellular level and to verify the hypothesis that infertility and the effect of SARS-CoV-2 on infertility may affect platelet mitochondrial bioenergetics and endogenous coenzyme Q10 levels.

Recruitment information / eligibility

• Status: Active, not recruiting
• Enrollment: 45
• Est. completion date: March 30, 2023
• Est. primary completion date: December 31, 2022
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: Male
• Age group: 18 Years to 40 Years

Eligibility

Inclusion Criteria:

• Infertile patients without COVID-19

• Infertile patients after COVID-19 Control group: healthy volunteers

Exclusion Criteria:

• disagreement with informed consent

Related Conditions & MeSH terms

• COVID-19
• Infertility
• Infertility, Male

Intervention

Other:
• diagnostic test and sperm analysis
Diagnostic Test: 2x14 ml of peripheral blood collected in the tube with anticoagulant, for platelet isolation, respirometry mitochondrial analysis, antioxidants (coenzyme Q10, vitamin E, gamma-tocopherol, beta-carotene) and TBARS estimation. Sperm analysis: standard spermiogram examination (volume, pH, number, motility and pathology) in the broker chamber, as well as extended examinations: mioxsys for redox potential, Vitalsperm (eosin-nigrosine staining) for sperm vitality and anti-sperm antibody (IgG) test.

Locations

Country	Name	City	State
Slovakia	Pharmacobiochemical Laboratory of Third Department of Internal Medicine, Faculty of Medicine Comenius University in Bratislava	Bratislava

Sponsors (2)

Lead Sponsor	Collaborator
Comenius University	GYN-FIV

Country where clinical trial is conducted

Slovakia, 

References & Publications (12)

Abobaker A, Raba AA. Does COVID-19 affect male fertility? World J Urol. 2021 Mar;39(3):975-976. doi: 10.1007/s00345-020-03208-w. Epub 2020 Apr 21. — View Citation

Boguenet M, Bouet PE, Spiers A, Reynier P, May-Panloup P. Mitochondria: their role in spermatozoa and in male infertility. Hum Reprod Update. 2021 Jun 22;27(4):697-719. doi: 10.1093/humupd/dmab001. Review. — View Citation

Gvozdjakova A, Klauco F, Kucharska J, Sumbalova Z. Is mitochondrial bioenergetics and coenzyme Q10 the target of a virus causing COVID-19? Bratisl Lek Listy. 2020;121(11):775-778. doi: 10.4149/BLL_2020_126. — View Citation

Gvozdjáková A, Kucharská J, Dubravicky J, Mojto V, Singh RB. Coenzyme Q10, a-tocopherol, and oxidative stress could be important metabolic biomarkers of male infertility. Dis Markers. 2015;2015:827941. doi: 10.1155/2015/827941. Epub 2015 Feb 25. — View Citation

Gvozdjakova A, Sumbalova Z, Kucharska J, Chladekova A, Rausova Z, Vancova O, Komlosi M, Ulicna O, Mojto V. Platelet mitochondrial bioenergetic analysis in patients with nephropathies and non-communicable diseases: a new method. Bratisl Lek Listy. 2019;120 — View Citation

Gvozdjáková A, Sumbalová Z, Kucharská J, Komlósi M, Rausová Z, Vancová O, Számošová M, Mojto V. Platelet Mitochondrial Respiration, Endogenous Coenzyme Q(10) and Oxidative Stress in Patients with Chronic Kidney Disease. Diagnostics (Basel). 2020 Mar 23;10 — View Citation

Gvozdjáková A, Sumbalová Z, Kucharská J, Szamosová M, Cápová L, Rausová Z, Vancová O, Mojto V, Langsjoen P, Palacka P. Platelet mitochondrial respiration and coenzyme Q10 could be used as new diagnostic strategy for mitochondrial dysfunction in rheumatoid — View Citation

Khalili MA, Leisegang K, Majzoub A, Finelli R, Panner Selvam MK, Henkel R, Mojgan M, Agarwal A. Male Fertility and the COVID-19 Pandemic: Systematic Review of the Literature. World J Mens Health. 2020 Oct;38(4):506-520. doi: 10.5534/wjmh.200134. Epub 2020 — View Citation

Pesta D, Gnaiger E. High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle. Methods Mol Biol. 2012;810:25-58. doi: 10.1007/978-1-61779-382-0_3. — View Citation

Rajak P, Roy S, Dutta M, Podder S, Sarkar S, Ganguly A, Mandi M, Khatun S. Understanding the cross-talk between mediators of infertility and COVID-19. Reprod Biol. 2021 Dec;21(4):100559. doi: 10.1016/j.repbio.2021.100559. Epub 2021 Sep 1. Review. — View Citation

Sheweita SA, Tilmisany AM, Al-Sawaf H. Mechanisms of male infertility: role of antioxidants. Curr Drug Metab. 2005 Oct;6(5):495-501. Review. — View Citation

Sumbalova Z, Kucharska J, Palacka P, Rausova Z, Langsjoen PH, Langsjoen AM, Gvozdjakova A. Platelet mitochondrial function and endogenous coenzyme Q10 levels are reduced in patients after COVID-19. Bratisl Lek Listy. 2022;123(1):9-15. doi: 10.4149/BLL_202 — View Citation

* Note: There are 12 references in all — Click here to view all references

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Clinical symptoms	Clinical symptoms: infertile patients without COVID-19 (vaccinated, or none vaccinated) Clinical symptoms patients after COVID-19	15 minutes
Primary	Damaged platelet mitochondrial bioenergetics 1	Basal oxygen consumption rate in intact platelets (ce)	1 day
Primary	Damaged platelet mitochondrial bioenergetics 2	rate of mitochondria LEAK respiration with CI-linked substrates (1PM - state 4)	1 day
Primary	Damaged platelet mitochondrial bioenergetics 3	CI-linked respiration coupled with ATPproduction (2D-CI-linked oxidative phosphorylation capacity), respiration after addition of cytochrome c (2C).	1 day
Primary	Damaged platelet mitochondrial bioenergetics 4	Maximal oxidative capacity (the electron transfer capacity -ET), after uncoupler titration (3U).	1 day
Primary	Damaged platelet mitochondrial bioenergetics 5	After addition of exogenous substrate glutamate (4G) non-coupled mitochondrial oxygen consumption.	1 day
Primary	Damaged platelet mitochondrial bioenergetics 6	Non-coupled oxygen consumption with CI&CII-linked substrate (5S) improvement of mitochondrial parameters representing OXPHOS- and electron tranport capacity (ET-capacity).	1 day
Primary	Endogenous coenzyme Q10-TOTAL 1	CoQ10-TOTAL in: Platelets (pmol.10-9 cells)	1 day
Primary	Endogenous coenzyme Q10-TOTAL 2	CoQ10-TOTAL in: Blood (µmol.L-1)	1 day
Primary	Endogenous coenzyme Q10-TOTAL 3	CoQ10-TOTAL in: Plasma (µmol.L-1)	1 day
Primary	Endogenous coenzyme TBARS	Endogenous concentration of CoQ10-TOTAL (ubiquinone + ubiquinol) in platelets, blood and plasma CoQ10-TOTAL in: TBARS in plasma (µmol.L-1).	1 day
Primary	Sperm analysis 1	standard spermiogram examination (volume, pH, number, motility and pathology) in the broker chamber	1 day
Primary	Sperm analysis 2	mioxsys for redox potential	1 day
Primary	Sperm analysis 3	Vitalsperm (eosin-nigrosine staining) for sperm vitality	1 day
Primary	Sperm analysis 4	anti-sperm antibody (IgG) test	1 day