Innate immune activation and mitochondrial ROS induce acute and persistent cardiac conduction system dysfunction after COVID-19
Deepthi Ashok, Ting Liu, Misato Nakanishi-Koakutsu, Joseph Criscione, Meghana Prakash, Alexis Tensfeldt, Byunggik Kim, Bryan Ho, Julian Chow, Morgan Craney, Mark J. Ranek, Brian L. Lin, Kyriakos Papanicolaou, Agnieszka Sidor, D. Brian Foster, Hee Cheol Cho, Andrew Pekosz, Jason Villano, Deok-Ho Kim, Brian O’Rourke
Deepthi Ashok, Ting Liu, Misato Nakanishi-Koakutsu, Joseph Criscione, Meghana Prakash, Alexis Tensfeldt, Byunggik Kim, Bryan Ho, Julian Chow, Morgan Craney, Mark J. Ranek, Brian L. Lin, Kyriakos Papanicolaou, Agnieszka Sidor, D. Brian Foster, Hee Cheol Cho, Andrew Pekosz, Jason Villano, Deok-Ho Kim, Brian O’Rourke
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Research Article Cardiology Immunology Infectious disease

Innate immune activation and mitochondrial ROS induce acute and persistent cardiac conduction system dysfunction after COVID-19

Abstract

Cardiac arrhythmias increase during acute SARS-CoV-2 infection and in long COVID syndrome, by unknown mechanisms. This study explored the acute and long-term effects of COVID-19 on cardiac electrophysiology and the cardiac conduction system (CCS) in a hamster model. Electrocardiograms and subpleural pressures were recorded by telemetry for 4 weeks after SARS-CoV-2 infection, and interferon-stimulated gene expression and macrophage infiltration of the CCS were assessed at 4 days and 4 weeks postinfection. COVID-19 induced pronounced tachypnea and cardiac arrhythmias, including bradycardia and persistent atrioventricular block, though no viral protein expression was detected in the heart. Arrhythmias developed rapidly, partially reversed, and then redeveloped, indicating persistent CCS injury. COVID-19 induced cardiac cytokine expression, connexin mislocalization, and CCS macrophage remodeling. Interestingly, sterile innate immune activation by direct cardiac injection of polyinosinic:polycytidylic acid (PIC) induced arrhythmias similar to those of COVID-19. PIC strongly induced cytokine secretion and interferon signaling in hearts, human induced pluripotent stem cell–derived cardiomyocytes, and engineered heart tissues, accompanied by alterations in excitation-contraction coupling. Importantly, the pulmonary and cardiac effects of COVID-19 were blunted by JAK/STAT inhibition or a mitochondrially targeted antioxidant, indicating that SARS-CoV-2 infection indirectly leads to arrhythmias by innate immune activation and redox stress, which could have implications for long COVID syndrome.

Authors

Deepthi Ashok, Ting Liu, Misato Nakanishi-Koakutsu, Joseph Criscione, Meghana Prakash, Alexis Tensfeldt, Byunggik Kim, Bryan Ho, Julian Chow, Morgan Craney, Mark J. Ranek, Brian L. Lin, Kyriakos Papanicolaou, Agnieszka Sidor, D. Brian Foster, Hee Cheol Cho, Andrew Pekosz, Jason Villano, Deok-Ho Kim, Brian O’Rourke

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Figure 6

JAK/STAT inhibition, but not mitochondrial antioxidant treatment, suppresses interferon responses in hiPSC-CM.

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JAK/STAT inhibition, but not mitochondrial antioxidant treatment, suppre...
(A) Schematic of molecular mechanisms of COVID-19 and dsRNA induced IFN response. Ruxolitinib suppresses IFN-stimulated response and mitoTEMPO suppresses mitochondrial damage and oxidative stress. (Created with BioRender.com.) (B and C) Inhibition of JAK/STAT signaling with ruxolitinib (Rux) (1 μM) significantly suppressed PIC-mediated induction of the interferon response signaling proteins STAT1, p-STAT1, IRF9, and MX1 in hiPSC-CM monolayers. MitoTEMPO (MT) (1 μM) does not significantly suppress the IFN response. Bar graphs show relative band intensity normalized to total protein loading (1-way ANOVA with Tukey’s multiple-comparison test). Lysates were from 3–5 different hiPSC-CM samples. (C) Inhibition of JAK/STAT signaling in hiPSC-derived engineered heart tissues (EHTs) with ruxolitinib, but not mitoTEMPO (MT), decreased the interferon response of signaling proteins STAT1, pSTAT1, IRF9, and MX1. Lysates loaded for the Western blot in C were from 3 different EHTs. (D) Cytokine secretion into the media induced by PIC, measured using a human Proteome Profiler array (inset shows exemplar blots), was greatly suppressed by Rux treatment in hiPSC-CM.