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. 2022 Apr 6;10(4):858.
doi: 10.3390/biomedicines10040858.

COVID-19 Infection Induce miR-371a-3p Upregulation Resulting in Influence on Male Fertility

Heike Goebel  1 , Barbara Koeditz  2 , Manuel Huerta  2 , Ersen Kameri  2 , Tim Nestler  2 , Thomas Kamphausen  3 , Johannes Friemann  4 , Matthias Hamdorf  5   6 , Timo Ohrmann  2 , Philipp Koehler  7   8 , Oliver A Cornely  7   8   9   10 , Manuel Montesinos-Rongen  11 , David Nicol  12 , Hubert Schorle  13 , Peter Boor  14 , Alexander Quaas  1 , Christian Pallasch  7 , Axel Heidenreich  2   15 , Melanie von Brandenstein  2
Affiliations

COVID-19 Infection Induce miR-371a-3p Upregulation Resulting in Influence on Male Fertility

Heike Goebel et al. Biomedicines. .

Abstract

In December 2019, the first case of COVID-19 was reported and since then several groups have already published that the virus can be present in the testis. To study the influence of SARS-CoV-2 which cause a dysregulation of the androgen receptor (AR) level, thereby leading to fertility problems and inducing germ cell testicular changes in patients after the infection. Formalin-Fixed-Paraffin-Embedded (FFPE) testicular samples from patients who died with or as a result of COVID-19 (n = 32) with controls (n = 6), inflammatory changes (n = 9), seminoma with/without metastasis (n = 11) compared with healthy biopsy samples (n = 3) were analyzed and compared via qRT-PCR for the expression of miR-371a-3p. An immunohistochemical analysis (IHC) and ELISA were performed in order to highlight the miR-371a-3p targeting the AR. Serum samples of patients with mild or severe COVID-19 symptoms (n = 34) were analyzed for miR-371a-3p expression. In 70% of the analyzed postmortem testicular tissue samples, a significant upregulation of the miR-371a-3p was detected, and 75% of the samples showed a reduced spermatogenesis. In serum samples, the upregulation of the miR-371a-3p was also detectable. The upregulation of the miR-371a-3p is responsible for the downregulation of the AR in SARS-CoV-2-positive patients, resulting in decreased spermatogenesis. Since the dysregulation of the AR is associated with infertility, further studies have to confirm if the identified dysregulation is regressive after a declining infection.

Keywords: COVID-19; SARS-CoV-2; androgen receptor; male infertility; miR-371a-3p.

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Conflict of interest statement

O.A.C. reports grants and personal fees from Actelion, Amplyx, Astellas, Basilea, Cidara, Da Volterra, F2G, Gilead, MedPace, Merck/MSD, Pfizer, Scynexis; grants from DFG (German Research Foundation), German Federal Ministry of Research and Education, Immunic, Janssen, Medicines Company, Melinta Therapeutics; personal fees from Allecra Therapeutics, Al-Jazeera Pharmaceuticals, Biocon, Biosys, CoRe Consulting, Entasis, Grupo Biotoscana, IQVIA, Matinas, Menarini, Molecular Partners, MSG-ERC, Mylan, Nabriva, Noxxon, Octapharma, Paratek, PSI, Roche Diagnostics, Seres, Shionogi; others from Wiley (Blackwell); outside the submitted work. P.K. is supported by the German Federal Ministry of Research and Education and the State of North Rhine-Westphalia, Germany and has received non-financial scientific grants from Miltenyi Biotec GmbH, Bergisch Gladbach, Germany, and the Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Cologne, Germany, and received lecture honoraria from and/or is advisor to Akademie für Infektionsmedizin e.V., Ambu GmbH, Astellas Pharma, European Confederation of Medical Mycology, Gilead Sciences, GPR Academy Ruesselsheim, MSD Sharp & Dohme GmbH, Noxxon N.V., Pfizer Pharma GmbH and University Hospital, LMU Munich outside the submitted work.

Figures

Figure 1
Figure 1
ACE2 in testicular FFPE samples from control (autopsy and testicular biopsy (TESE)) and COVID-19 patients. In all three depicted samples, ACE2 expression could be found. Shown here is a representative example out of five (40× magnification). Insert shows respective negative controls (63×).
Figure 2
Figure 2
Upper part: IHC from testicular tissue samples. (A,B) present a slightly reduced spermatogenesis and normal AR expression in healthy control tissue sample (brownish marked nuclei, clone AR441) (C,D) Example of COVID-19 positive patient with no expression of the AR as well as a reduced spermatogenesis. (E,F) are examples of inflammatory changes control and (G,H) from autopsy of a case of previous intensive care treatment. (I,J) autopsy of vaccinated man dying of COVID-19. Figures exemplified a collective of each analyzed group used. Left side H&E (spermiogenesis yellow arrowheads), right side IHC for androgen receptor AR441 (black arrowheads), each at 400× magnification. Lower part: Androgen receptor ELISA from FFPE tissue. Total protein was isolated from FFPE tissue and 1 µg of total protein was used for the analysis by ELISA. Significant decreased AR values were found in testicular tissue samples compared with control group. *** p < 0.0001, ** p < 0.001, unmarked = ns. Control ELISA for ß-actin to control isolation quantity (K). Detection of AR protein in FFPE samples (L). Significant reduction in the AR levels in COVID-19 samples (±30%) compared to control autopsy and ICU non COVID-19 were detectable (*** p < 0.001).
Figure 2
Figure 2
Upper part: IHC from testicular tissue samples. (A,B) present a slightly reduced spermatogenesis and normal AR expression in healthy control tissue sample (brownish marked nuclei, clone AR441) (C,D) Example of COVID-19 positive patient with no expression of the AR as well as a reduced spermatogenesis. (E,F) are examples of inflammatory changes control and (G,H) from autopsy of a case of previous intensive care treatment. (I,J) autopsy of vaccinated man dying of COVID-19. Figures exemplified a collective of each analyzed group used. Left side H&E (spermiogenesis yellow arrowheads), right side IHC for androgen receptor AR441 (black arrowheads), each at 400× magnification. Lower part: Androgen receptor ELISA from FFPE tissue. Total protein was isolated from FFPE tissue and 1 µg of total protein was used for the analysis by ELISA. Significant decreased AR values were found in testicular tissue samples compared with control group. *** p < 0.0001, ** p < 0.001, unmarked = ns. Control ELISA for ß-actin to control isolation quantity (K). Detection of AR protein in FFPE samples (L). Significant reduction in the AR levels in COVID-19 samples (±30%) compared to control autopsy and ICU non COVID-19 were detectable (*** p < 0.001).
Figure 3
Figure 3
miR 371a-3p in FFPE testicular tissue samples from COVID-19 patients compared to patients with ICU/ECMO, inflamed testis and Seminoma patients without metastasis (M−) or with metastasis (M+). All samples were normalized to 5s rRNA and corresponding controls (for COVID-19 and ICU controls from autopsy patients and for inflamed testis and Seminoma controls from testicular surgery without inflammatory disease or tumor). An increased expression, comparable with the miR expression of Seminoma patients with metastasis was detectable. In case of ICU non COVID-19, Seminoma without metastasis and inflamed, a significant reduction compared to COVID-19 was detectable. *** p < 0.0001, ** p < 0.001, * p < 0.01, unmarked = ns.
Figure 4
Figure 4
IHC followed by miR-371a-3p in situ PCR was performed. As it can be seen, those samples (controls, autopsy and TESE) showing high AR levels (here seen in brownish coloration of the nuclei, 40×) the miR-371a-3p expression is reduced (FITC staining right row, 40×, inserts 63×). DAPI staining was included to visualize the nuclei. In COVID-19 samples were the AR levels were reduced (nuclei appears blue, first row) the miR-371a-3p was significantly upregulated (last row). Here, results out of three independent experiments are presented.
Figure 5
Figure 5
qRT-PCR results for the detection of miR-371a-3p in serum from patients with mild or severe COVID-19. (A) depicted the miR-371a-3p expression female and male. Upregulation was found in case of male patients either with mild or severe symptoms. (B) summarized all female and male cases showing the expression of the miR-371a-3p. Increase in male patients was detectable (* p < 0.01).
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References

    1. Zhang Y., Geng X., Tan Y., Li Q., Xu C., Xu J., Hao L., Zeng Z., Luo X., Liu F., et al. New understanding of the damage of SARS-CoV-2 infection outside the respiratory system. Biomed. Pharmacother. 2020;127:110195. doi: 10.1016/j.biopha.2020.110195. - DOI - PMC - PubMed
    1. Kirtipal N., Kumar S., Dubey S.K., Dwivedi V.D., Gireesh Babu K., Maly P., Bharadwaj S. Understanding on the possible routes for SARS CoV-2 invasion via ACE2 in the host linked with multiple organs damage. Infect. Genet. Evol. 2022;99:105254. doi: 10.1016/j.meegid.2022.105254. - DOI - PMC - PubMed
    1. Salamanna F., Maglio M., Landini M.P., Fini M. Body Localization of ACE-2: On the Trail of the Keyhole of SARS-CoV-2. Front. Med. 2020;7:594495. doi: 10.3389/fmed.2020.594495. - DOI - PMC - PubMed
    1. Ejaz H., Alsrhani A., Zafar A., Javed H., Junaid K., Abdalla A.E., Abosalif K.O.A., Ahmed Z., Younas S. COVID-19 and comorbidities: Deleterious impact on infected patients. J. Infect. Public Health. 2020;13:1833–1839. doi: 10.1016/j.jiph.2020.07.014. - DOI - PMC - PubMed
    1. Migliore L., Nicoli V., Stoccoro A. Gender Specific Differences in Disease Susceptibility: The Role of Epigenetics. Biomedicines. 2021;9:652. doi: 10.3390/biomedicines9060652. - DOI - PMC - PubMed