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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

Liu et al., Stem Cell Research & Therapy, doi:10.1186/s13287-023-03485-3
Sep 2023  
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Ivermectin for COVID-19
4th treatment shown to reduce risk in August 2020
 
*, now with p < 0.00000000001 from 104 studies, recognized in 23 countries.
No treatment is 100% effective. Protocols combine treatments. * >10% efficacy, ≥3 studies.
4,400+ studies for 79 treatments. c19ivm.org
In Vitro study showing that ivermectin and meclizine mitigated cardiac cell death and dysfunction caused by SARS-CoV-2 viral genes.
Authors found that SARS-CoV-2 viral genes Nsp6, Nsp8, and M had harmful effects on human cardiomyocytes (heart muscle cells) derived from human pluripotent stem cells.
SARS-CoV-2 infection and overexpression of these genes depleted cellular ATP, activated genes related to cell death and inflammation, suppressed genes important for heart contraction, and compromised calcium handling.
Ivermectin and meclizine restored ATP levels and mitigated the negative effects of the SARS-CoV-2 viral genes.
68 preclinical studies support the efficacy of ivermectin for COVID-19:
Ivermectin, better known for antiparasitic activity, is a broad spectrum antiviral with activity against many viruses including H7N766, Dengue32,67,68, HIV-168, Simian virus 4069, Zika32,70,71, West Nile71, Yellow Fever72,73, Japanese encephalitis72, Chikungunya73, Semliki Forest virus73, Human papillomavirus52, Epstein-Barr52, BK Polyomavirus74, and Sindbis virus73.
Ivermectin inhibits importin-α/β-dependent nuclear import of viral proteins66,68,69,75, shows spike-ACE2 disruption at 1nM with microfluidic diffusional sizing33, binds to glycan sites on the SARS-CoV-2 spike protein preventing interaction with blood and epithelial cells and inhibiting hemagglutination36,76, shows dose-dependent inhibition of wildtype and omicron variants31, exhibits dose-dependent inhibition of lung injury56,61, may inhibit SARS-CoV-2 via IMPase inhibition32, may inhibit SARS-CoV-2 induced formation of fibrin clots resistant to degradation5, inhibits SARS-CoV-2 3CLpro49, may inhibit SARS-CoV-2 RdRp activity24, may minimize viral myocarditis by inhibiting NF-κB/p65-mediated inflammation in macrophages55, may be beneficial for COVID-19 ARDS by blocking GSDMD and NET formation77, may inhibit SARS-CoV-2 by disrupting CD147 interaction78-81, shows protection against inflammation, cytokine storm, and mortality in an LPS mouse model sharing key pathological features of severe COVID-1954,82, may be beneficial in severe COVID-19 by binding IGF1 to inhibit the promotion of inflammation, fibrosis, and cell proliferation that leads to lung damage4, may minimize SARS-CoV-2 induced cardiac damage35,43, increases Bifidobacteria which play a key role in the immune system83, has immunomodulatory46 and anti-inflammatory65,84 properties, and has an extensive and very positive safety profile85.
Liu et al., 13 Sep 2023, peer-reviewed, 7 authors. Contact: juliliu@outlook.com, lyang7@iu.edu.
In Vitro studies are an important part of preclinical research, however results may be very different in vivo.
This PaperIvermectinAll
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. Competing interests The authors declare no competing interests. • fast, convenient online submission • thorough peer review by experienced researchers in your field • rapid publication on acceptance • support for research data, including large and complex data types • gold Open Access which fosters wider collaboration and increased citations maximum visibility for your research: over 100M website views per year • At BMC, research is always in progress. Learn more biomedcentral.com/submissions Ready to submit your research..
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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.</jats:p>\n' ' </jats:sec><jats:sec>\n' ' <jats:title>Methods</jats:title>\n' ' <jats:p>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. 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