SARS-CoV-2 viral genes Nsp6, Nsp8, and M compromise cellular ATP levels to impair survival and function of human pluripotent stem cell-derived cardiomyocytes
Juli Liu, Shiyong Wu, Yucheng Zhang, Cheng Wang, Sheng Liu, Jun Wan, Lei Yang
Stem Cell Research & Therapy, doi:10.1186/s13287-023-03485-3
Background Cardiovascular complications significantly augment the overall COVID-19 mortality, largely due to the susceptibility of human cardiomyocytes (CMs) to SARS-CoV-2 virus. SARS-CoV-2 virus encodes 27 genes, whose specific impacts on CM health are not fully understood. This study elucidates the deleterious effects of SARS-CoV-2 genes Nsp6, M, and Nsp8 on human CMs. Methods CMs were derived from human pluripotent stem cells (hPSCs), including human embryonic stem cells and induced pluripotent stem cells, using 2D and 3D differentiation methods. We overexpressed Nsp6, M, or Nsp8 in hPSCs and then applied whole mRNA-seq and mass spectrometry for multi-omics analysis. Co-immunoprecipitation mass spectrometry was utilized to map the protein interaction networks of Nsp6, M, and Nsp8 within host hiPSC-CMs.
Results Nsp6, Nsp8, and M globally perturb the transcriptome and proteome of hPSC-CMs. SARS-CoV-2 infection and the overexpression of Nsp6, Nsp8, or M coherently upregulated genes associated with apoptosis and immune/ inflammation pathways, whereas downregulated genes linked to heart contraction and functions. Global interactome analysis revealed interactions between Nsp6, Nsp8, and M with ATPase subunits. Overexpression of Nsp6, Nsp8, or M significantly reduced cellular ATP levels, markedly increased apoptosis, and compromised Ca 2+ handling in hPSC-CMs. Importantly, administration of FDA-approved drugs, ivermectin and meclizine, could restore ATP levels, thereby mitigating apoptosis and dysfunction in hPSC-CMs overexpressing Nsp6, Nsp8, or M.
Conclusion Overall, our findings uncover the extensive damaging effects of Nsp6, Nsp8, and M on hPSC-CMs, underlining the crucial role of ATP homeostasis in CM death and functional abnormalities induced by these SARS-CoV-2 genes, and reveal the potential therapeutic strategies to alleviate these detrimental effects with FDA-approved drugs.
Supplementary Information The online version contains supplementary material available at https:// doi. org/ 10. 1186/ s13287-023-03485-3. Additional file 1. Supplementary Figures. S1 . mRNA expression profiling of Nsp6OE, Nsp8OE, or MOE and control hESC-CMs.
Additional file 2. Table Additional file 3. Table S2 . Protein expression profiling of Nsp6OE, Nsp8OE, or MOE and control hESC-CMs. Additional file 4. Table S3 . CoIP-MS of Nsp6OE, Nsp8OE, or MOE and control hESC-CMs. Additional file 5. Table S4 . Primers for RT-qPCR. Author contributions JL initiated and designed the studies. JL and SW performed all experiments, and data collection and analyses. YZ, SL, and JW conducted bioinformatics analyses. CW assisted in CM functional studies. JL, SW, JW, and LY wrote the manuscript. All authors read and approved the final manuscript.
Declarations Ethics approval and consent to participate Not applicable.
Consent for publication Not applicable.
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