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SCIENCE CHINA Life Sciences, Volume 63 , Issue 7 : 1006-1015(2020) https://doi.org/10.1007/s11427-020-1705-0

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

Xixi Liu 1,2,4,†, Yidong Chen 1,2,4,5,†, Wenhao Tang 6, Li Zhang 2, Wei Chen 1,2,4,5, Zhiqiang Yan 1,2,4,5, Peng Yuan 2,3,4, Ming Yang 1,2,4,5, Siming Kong 1,2,4,5, Liying Yan 1,2,3,4,*, Jie Qiao 1,2,3,4,5,*
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  • ReceivedMar 31, 2020
  • AcceptedApr 21, 2020
  • PublishedApr 30, 2020

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 single-cell 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.


Acknowledgment

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


Interest statement

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


Supplement

SUPPORTING INFORMATION

Figure S1 Classical testis cell markers expression.

Figure S2 Expression pattern of ACE2 related genes.

Figure S3 The cell number and classical marker expression of Sertoli cells.

The supporting information is available online at http://life.scichina.com and https://link.springer.com. The supporting materials are published as submitted, without typesetting or editing. The responsibility for scientific accuracy and content remains entirely with the authors.


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  • Figure 1

    ACE2 expression pattern and the ratio of ACE2-positive cells in normal testis cells. A, T-distributed stochastic neighbor embedding (t-SNE) plot of all normal 2,854 single cells in the previous paper. Different colors represent different cell types. B, The expression levels (TPM) of ACE2 in all cell types. C, Cell type distribution ratio of ACE2-positive cells. Different colors represent different cell types. D, ACE2-positive cells ratio in each cell type. E, ACE2-positive cells ratio and ACE2 average expression levels (TPM) in different age groups.

  • Figure 2

    Different ACE2 expression levels in normal human male testis cells. A, Violin plot of five classes with different ACE2 expression levels. B, Five classes composition distribution in all ACE2-positive cell types. C, Gene ontology terms of different classes. D, Venn plot of intersection genes in different classes. E, Heatmap of intersection genes in all classes. Class F represents ACE2-negative cells; Class A represents cells with ACE2 expression in the range of 0 to 20; Class B represents cells with ACE2 expression in the range of 20 to 100; Class C represents cells with ACE2 expression in the range of 100 to 150; Class D represents cells with ACE2 expression in the range of 150 to 300; Class E represents cells with ACE2 expression levels greater than 300. F, Lineplots of increasing genes. Class F represents ACE2-negative cells; Class A represents cells with ACE2 expression in the range of 0 to 20; Class B represents cells with ACE2 expression in the range of 20 to 100; Class C represents cells with ACE2 expression in the range of 100 to 150; Class D represents cells with ACE2 expression in the range of 150 to 300; Class E represents cells with ACE2 expression levels greater than 300.

  • Figure 3

    Possible COVID-19 process-related genes expression pattern in normal testis cells. A, The expression levels (TPM) of TMPRSS2 in all cell types. B, Lineplot of ACE2 and TMPRSS2. C, Lineplot of CTSL and CTSB. D, Lineplot of BSG.

  • Figure 4

    ACE2 expression pattern in NOA patients. A, ACE2-positive cells ratio in each Sertoli cell group. Nor.ST refers to Sertoli cells from normal data. NOA.ST refers to Sertoli cells from three non-obstructive azoospermia data. B, Five classes composition distribution in each Sertoli cell group. C, The expression levels of ACE2 in different Sertoli cell groups. D, The expression levels of TMPRSS2 in different Sertoli cell groups. E, Gene ontology terms of DEGs in different classes of Sertoli cells from non-obstructive azoospermia data. DEGs refer to differentially expressed genes. F, Gene ontology terms of DEGs in different classes of Sertoli cells from non-obstructive azoospermia and normal data. DEGs refer to differentially expressed genes. G, The average expression levels of up-regulated gene lists in normal E, F class and NOA E, F class. H, The average expression levels of down-regulated gene list in normal E, F class and NOA E, F class.

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