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Single-cell transcriptome analysis of the novel coronavirus (SARS-CoV-2) associated gene ACE2 expression in normal and non-obstructive azoospermia (NOA) human male testes

Abstract

Being infected by SARS-CoV-2 may cause damage to multiple organs in patients, such as the lung, liver and heart. Angiotensin-converting enzyme 2 (ACE2), reported as a SARS-CoV-2 receptor, is also expressed in human male testes. This suggests a potential risk in human male reproductive system. However, the characteristics of ACE2-positive cells and the expression of other SARS-CoV-2 process-related genes are still worthy of further investigation. Here, we performed singlecell RNA seq (scRNA-seq) analysis on 853 male embryo primordial germ cells (PGCs) and 2,854 normal testis cells to assess the effects of the SARS-CoV-2 virus on the male reproductive system from embryonic stage to adulthood. We also collected and constructed the scRNA-seq library on 228 Sertoli cells from three non-obstructive azoospermia (NOA) patients to assess the effects at disease state. We found that ACE2 expressing cells existed in almost all testis cell types and Sertoli cells had highest expression level and positive cells ratio. Moreover, ACE2 was also expressed in human male PGCs. In adulthood, the level of ACE2 expression decreased with the increase of age. We also found that ACE2 positive cells had high expressions of stress response and immune activation-related genes. Interestingly, some potential SARS-CoV-2 process-related genes such as TMPRSS2, BSG, CTSL and CTSB had different expression patterns in the same cell type. Furthermore, ACE2 expression level in NOA donors’ Sertoli cells was significantly decreased. Our work would help to assess the risk of SARS-CoV-2 infection in the male reproductive system.

References

  1. Anders, S., Pyl, P.T., and Huber, W. (2014). HTSeq—a Python framework to work with high-throughput sequencing data. Bioinformatics 31, 166–169.

    Article  Google Scholar 

  2. Backhaus, S., Zakrzewicz, A., Richter, K., Damm, J., Wilker, S., Fuchs-Moll, G., Küllmar, M., Hecker, A., Manzini, I., Ruppert, C., et al. (2017). Surfactant inhibits ATP-induced release of interleukin-1β via nicotinic acetylcholine receptors. J Lipid Res 58, 1055–1066.

    CAS  Article  Google Scholar 

  3. Ding, Y., He, L., Zhang, Q., Huang, Z., Che, X., Hou, J., Wang, H., Shen, H., Qiu, L., Li, Z., et al. (2004). Organ distribution of severe acute respiratory syndrome (SARS) associated coronavirus (SARS-CoV) in SARS patients: implications for pathogenesis and virus transmission pathways. J Pathol 203, 622–630.

    CAS  Article  Google Scholar 

  4. Douglas, G.C., O’Bryan, M.K., Hedger, M.P., Lee, D.K.L., Yarski, M.A., Smith, A.I., and Lew, R.A. (2004). The novel angiotensin-converting enzyme (ACE) homolog, ACE2, is selectively expressed by adult Leydig cells of the testis. Endocrinology 145, 4703–4711.

    CAS  Article  Google Scholar 

  5. Hoffmann, M., Kleine-Weber, H., Schroeder, S., Krüger, N., Herrler, T., Erichsen, S., Schiergens, T.S., Herrler, G., Wu, N.H., Nitsche, A., et al. (2020). SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell 181, 271–280.e8.

    CAS  Article  Google Scholar 

  6. Koguchi, T., Tanikawa, C., Mori, J., Kojima, Y., and Matsuda, K. (2016). Regulation of myo-inositol biosynthesis by p53-ISYNA1 pathway. Int J Oncol 48, 2415–2424.

    CAS  Article  Google Scholar 

  7. Li, L., Dong, J., Yan, L., Yong, J., Liu, X., Hu, Y., Fan, X., Wu, X., Guo, H., Wang, X., et al. (2017). Single-cell RNA-seq analysis maps development of human germline cells and gonadal niche interactions. Cell Stem Cell 20, 858–873.e4.

    CAS  Article  Google Scholar 

  8. Liu, J., Ji, H., Zheng, W., Wu, X., Zhu, J.J., Arnold, A.P., and Sandberg, K. (2010). Sex differences in renal angiotensin converting enzyme 2 (ACE2) activity are 17β-oestradiol-dependent and sex chromosome-independent. Biol Sex Dif 1, 6.

    CAS  Article  Google Scholar 

  9. Lu, R., Zhao, X., Li, J., Niu, P., Yang, B., Wu, H., Wang, W., Song, H., Huang, B., Zhu, N., et al. (2020). Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding. Lancet 395, 565–574.

    CAS  Article  Google Scholar 

  10. Ma, L., Xie, W., Li, D., Shi, L., Mao, Y., Xiong, Y., Zhang, Y., and Zhang, M. (2020). Effect of SARS-CoV-2 infection upon male gonadal function: A single center- based study. medRxiv, https://doi.org/10.1101/2020.03.21.20037267.

  11. Qi, L., Chi, X., Xu, J., Yang, J., Gong, E., and Gu, J. (2007). Pathological changes of testis tissue in SARS patients. Basic Clin Med.

  12. Qi, S., Huang, M., Teng, H., Lu, Y., Jiang, M., Wang, L., Shi, J., Ma, Q., Gu, G., Xin, Y., et al. (2015). Autoantibodies to chromogranin A are potential diagnostic biomarkers for non-small cell lung cancer. Tumor Biol 36, 9979–9985.

    CAS  Article  Google Scholar 

  13. Reis, A.B., Araújo, F.C., Pereira, V.M., Dos Reis, A.M., Santos, R.A., and Reis, F.M. (2010). Angiotensin (1–7) and its receptor Mas are expressed in the human testis: implications for male infertility. J Mol Hist 41, 75–80.

    CAS  Article  Google Scholar 

  14. Satija, R., Farrell, J.A., Gennert, D., Schier, A.F., and Regev, A. (2015). Spatial reconstruction of single-cell gene expression data. Nat Biotechnol 33, 495–502.

    CAS  Article  Google Scholar 

  15. Shen, Q., Xiao, X., Aierken, A., Liao, M., and Hua, J. (2020). The ACE2 expression in Sertoli cells and Germ cells may cause male reproductive disorder after SARS-CoV-2 infection. Preprint, https://doi.org/10.31219/osf.io/fs5hd.

  16. Simmons, G., Gosalia, D.N., Rennekamp, A.J., Reeves, J.D., Diamond, S. L., and Bates, P. (2005). Inhibitors of cathepsin L prevent severe acute respiratory syndrome coronavirus entry. Proc Natl Acad Sci USA 102, 11876–11881.

    CAS  Article  Google Scholar 

  17. Trapnell, C., Pachter, L., and Salzberg, S.L. (2009). TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25, 1105–1111.

    CAS  Article  Google Scholar 

  18. Wang, K., Chen, W., Zhou, Y.S., Lian, J.Q., Zhang, Z., Du, P., Gong, L., Zhang, Y., Cui, H.Y., Geng, J.J., et al. (2020). SARS-CoV-2 invades host cells via a novel route: CD147-spike protein. bioRxiv, https://doi.org/10.1101/2020.03.14.988345.

  19. Wang, M., Liu, X., Chang, G., Chen, Y., An, G., Yan, L., Gao, S., Xu, Y., Cui, Y., Dong, J., et al. (2018). Single-cell RNA sequencing analysis reveals sequential cell fate transition during human spermatogenesis. Cell Stem Cell 23, 599–614.e4.

    CAS  Article  Google Scholar 

  20. Wang, Z., and Xu, X. (2020). scRNA-seq profiling of human testes reveals the presence of the ACE2 receptor, a target for SARS-CoV-2 infection in spermatogonia, Leydig and Sertoli cells. Cells 9, 920–928.

    Article  Google Scholar 

  21. Williamson, C.R., Khurana, S., Nguyen, P., Byrne, C.J., and Tai, T.C. (2017). Comparative analysis of renin-angiotensin system (RAS)-related gene expression between hypertensive and normotensive rats. Med Sci Monit Basic Res 23, 20–24.

    Article  Google Scholar 

  22. Zou, X., Chen, K., Zou, J., Han, P., Hao, J., and Han, Z. (2020). Single-cell RNA-seq data analysis on the receptor ACE2 expression reveals the potential risk of different human organs vulnerable to 2019-nCoV infection. Front Med https://doi.org/10.1007/s11684-020-0754-0.

  23. Zuo, X., Xu, W., Xu, M., Tian, R., Moussalli, M.J., Mao, F., Zheng, X., Wang, J., Morris, J.S., Gagea, M., et al. (2017). Metastasis regulation by PPARD expression in cancer cells. JCI Insight 2, e91419.

    Article  Google Scholar 

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Acknowledgements

This work was funded by Beijing Municipal Science & Technology Commission (Z191100006619075) and the National Natural Science Foundation of China (81521002 and 81730038).

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Correspondence to Liying Yan or Jie Qiao.

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Compliance and ethics The author(s) declare that they have no conflict of interest.

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Liu, X., Chen, Y., Tang, W. et al. Single-cell transcriptome analysis of the novel coronavirus (SARS-CoV-2) associated gene ACE2 expression in normal and non-obstructive azoospermia (NOA) human male testes. Sci. China Life Sci. 63, 1006–1015 (2020). https://doi.org/10.1007/s11427-020-1705-0

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Keywords

  • SARS-CoV-2
  • COVID-19
  • testis cells
  • ACE2
  • TMPRSS2
  • fetal
  • adult
  • normal
  • NOA