Inhibition of SARS-CoV-2 infection in human iPSC-derived cardiomyocytes by targeting the Sigma-1 receptor disrupts cytoarchitecture and beating

SARS-CoV-2 infects cardiac cells and causes heart dysfunction. Conditions such as myocarditis and arrhythmia have been reported in COVID-19 patients. The Sigma-1 receptor (S1R) is a ubiquitously expressed chaperone that plays a central role in cardiomyocyte function. S1R has been proposed as a therapeutic target because it may affect SARS-CoV-2 replication; however, the impact of the inhibition of S1R in human cardiomyocytes remains to be described. In this study, we investigated the consequences of S1R inhibition in iPSC-derived human cardiomyocytes (hiPSC-CM). SARS-CoV-2 infection in hiPSC-CM was productive and reduced cell survival. S1R inhibition decreased both the number of infected cells and viral particles after 48 hours. S1R inhibition also prevented the release of pro-inflammatory cytokines and cell death. Although the S1R antagonist NE-100 triggered those protective effects, it compromised cytoskeleton integrity by downregulating the expression of structural-related genes and reducing beating frequency. Our findings suggest that the detrimental effects of S1R inhibition in human cardiomyocytes’ integrity may abrogate its therapeutic potential against COVID and should be carefully considered.

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The Protective Effects of Dexmedetomidine against Hypoxia/Reoxygenation-Induced Inflammatory Injury and Permeability in Brain Endothelial Cells Mediated by Sigma-1 Receptor

ACS Chemical Neuroscience ◽

10.1021/acschemneuro.1c00032 ◽

2021 ◽

Author(s):

Qin Zhao ◽

Shoushui Yu ◽

Yong Ling ◽

Shiyuan Hao ◽

Jia Liu

Keyword(s):

Endothelial Cells ◽

Protective Effects ◽

Brain Endothelial Cells ◽

Inflammatory Injury ◽

Sigma 1 Receptor ◽

Sigma 1 ◽

Hypoxia Reoxygenation

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Flow Cytometry Analysis of HIV-1 Env Conformations at the Surface of Infected Cells and Virions: Role of Nef, CD4, and SERINC5

Journal of Virology ◽

10.1128/jvi.01783-19 ◽

2019 ◽

Vol 94 (6) ◽

Cited By ~ 3

Author(s):

Isabelle Staropoli ◽

Jérémy Dufloo ◽

Anaïs Ducher ◽

Pierre-Henri Commere ◽

Anna Sartori-Rupp ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Cellular Proteins ◽

Viral Infectivity ◽

Viral Particles ◽

Immunodeficiency Virus ◽

Infected Cells ◽

Env Protein ◽

The Impact ◽

Hiv 1

ABSTRACT The HIV-1 Env protein is exposed at the surface of virions and infected cells. Env fluctuates between different closed and open structural states and these conformations influence both viral infectivity and sensitivity to antibody binding and neutralization. We established a flow virometry assay to visualize Env proteins at the surface of human immunodeficiency virus type 1 (HIV-1) virions. The assay is performed on ultracentrifuged fluorescent viral particles that are stained with a panel of broadly neutralizing antibodies (bNAbs) and nonneutralizing antibodies (nnAbs) that probe different epitopes of Env. We used this assay to compare Env at the surface of producer cells and viral particles and to analyze the effect of Nef, CD4, and SERINC5 on Env accessibility to antibodies. We studied the laboratory-adapted strain NL4-3 and two transmitted/founder viruses, THRO and CH058. We confirm that antibody accessibility varies between viral strains and show that Nef, CD4, and SERINC5 additively impact Env conformations. We further demonstrate that the Env accessibility profile on virions is globally similar to that observed on HIV-1-infected cells, with some noticeable differences. For instance, nnAbs bind to virions more efficiently than to producer cells, likely reflecting changes in Env conformational states on mature viral particles. This test complements other techniques and provides a convenient and simple tool for quantifying and probing the structure of Env at the virion surface and to analyze the impact of viral and cellular proteins on these parameters. IMPORTANCE HIV-1 Env conformation is one of the key parameters determining viral infectivity. The flow virometry-based assay developed in this study allows for the characterization of proteins incorporated in HIV-1 particles. We studied the conformation of HIV-1 Env and the impact that the viral protein Nef and the cellular proteins CD4 and SERINC5 have on Env accessibility to antibodies. Our assay permitted us to highlight some noticeable differences in the conformation of Env between producer cells and viral particles. It contributes to a better understanding of the actual composition of HIV-1 particles.

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Protective effect of creatine against inhibition by methylglyoxal of mitochondrial respiration of cardiac cells

Biochemical Journal ◽

10.1042/bj20021576 ◽

2003 ◽

Vol 372 (2) ◽

pp. 661-669 ◽

Cited By ~ 20

Author(s):

Soumya Sinha ROY ◽

Swati BISWAS ◽

Manju RAY ◽

Subhankar RAY

Keyword(s):

Creatine Kinase ◽

Protective Effect ◽

Mitochondrial Respiration ◽

Therapeutic Potential ◽

Ehrlich Ascites Carcinoma ◽

Creatine Phosphate ◽

Inhibitory Effect ◽

Cardiac Cells ◽

Protective Effects ◽

Tissue Slices

Previous publications from our laboratory have shown that methylglyoxal inhibits mitochondrial respiration of malignant and cardiac cells, but it has no effect on mitochondrial respiration of other normal cells [Biswas, Ray, Misra, Dutta and Ray (1997) Biochem. J. 323, 343–348; Ray, Biswas and Ray (1997) Mol. Cell. Biochem. 171, 95–103]. However, this inhibitory effect of methylglyoxal is not significant in cardiac tissue slices. Moreover, post-mitochondrial supernatant (PMS) of cardiac cells could almost completely protect the mitochondrial respiration against the inhibitory effect of methylglyoxal. A systematic search indicated that creatine present in cardiac cells is responsible for this protective effect. Glutathione has also some protective effect. However, creatine phosphate, creatinine, urea, glutathione disulphide and β-mercaptoethanol have no protective effect. The inhibitory and protective effects of methylglyoxal and creatine respectively on cardiac mitochondrial respiration were studied with various concentrations of both methylglyoxal and creatine. Interestingly, neither creatine nor glutathione have any protective effect on the inhibition by methylglyoxal on the mitochondrial respiration of Ehrlich ascites carcinoma cells. The creatine and glutathione contents of several PMS, which were tested for the possible protective effect, were measured. The activities of two important enzymes, namely glyoxalase I and creatine kinase, which act upon glutathione plus methylglyoxal and creatine respectively, were also measured in different PMS. Whether mitochondrial creatine kinase had any role in the protective effect of creatine had also been investigated using 1-fluoro-2,4-dinitrobenzene, an inhibitor of creatine kinase. The differential effect of creatine on mitochondria of cardiac and malignant cells has been discussed with reference to the therapeutic potential of methylglyoxal.

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Inhibition of SARS-CoV-2 infection in human cardiomyocytes by targeting the Sigma-1 receptor disrupts cytoskeleton architecture and contractility

10.1101/2021.02.20.432092 ◽

2021 ◽

Author(s):

José Alexandre Salerno ◽

Thayana Torquato ◽

Jairo R. Temerozo ◽

Livia Goto-Silva ◽

Mayara Mendes ◽

...

Keyword(s):

Therapeutic Strategy ◽

Sigma 1 Receptor ◽

Human Cardiomyocytes ◽

Sigma 1 ◽

Induced Pluripotent ◽

And Function ◽

Beating Frequency ◽

Cytoskeleton Integrity

ABSTRACTHeart dysfunction, represented by conditions such as myocarditis and arrhythmia, has been reported in COVID-19 patients. Therapeutic strategies focused on the cardiovascular system, however, remain scarce. The Sigma-1 receptor (S1R) has been recently proposed as a therapeutic target because its inhibition reduces SARS-CoV-2 replication. To investigate the role of S1R in SARS-CoV-2 infection in the heart, we used human cardiomyocytes derived from induced pluripotent stem cells (hiPSC-CM) as an experimental model. Here we show that the S1R antagonist NE-100 decreases SARS-CoV-2 infection and viral replication in hiPSC-CMs. Also, NE-100 reduces cytokine release and cell death associated with infection. Because S1R is involved in cardiac physiology, we investigated the effects of NE-100 in cardiomyocyte morphology and function. We show that NE-100 compromises cytoskeleton integrity and reduces beating frequency, causing contractile impairment. These results show that targeting S1R to challenge SARS-CoV-2 infection may be a useful therapeutic strategy but its detrimental effects in vivo on cardiac function should not be ignored.

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Sigma-1 Receptor Is Critical for Mitochondrial Activity and Unfolded Protein Response in Larval Zebrafish

International Journal of Molecular Sciences ◽

10.3390/ijms222011049 ◽

2021 ◽

Vol 22 (20) ◽

pp. 11049

Author(s):

Lucie Crouzier ◽

Morgane Denus ◽

Elodie M. Richard ◽

Amarande Tavernier ◽

Camille Diez ◽

...

Keyword(s):

Er Stress ◽

Unfolded Protein Response ◽

Transmembrane Protein ◽

Critical Role ◽

Mitochondrial Activity ◽

Sigma 1 Receptor ◽

Unfolded Protein ◽

Sigma 1 ◽

Protein Response ◽

The Impact

The sigma-1 receptor (S1R) is a highly conserved transmembrane protein highly enriched in mitochondria-associated endoplasmic reticulum (ER) membranes, where it interacts with several partners involved in ER-mitochondria Ca2+ transfer, activation of the ER stress pathways, and mitochondria function. We characterized a new S1R deficient zebrafish line and analyzed the impact of S1R deficiency on visual, auditory and locomotor functions. The s1r+25/+25 mutant line showed impairments in visual and locomotor functions compared to s1rWT. The locomotion of the s1r+25/+25 larvae, at 5 days post fertilization, was increased in the light and dark phases of the visual motor response. No deficit was observed in acoustic startle response. A critical role of S1R was shown in ER stress pathways and mitochondrial activity. Using qPCR to analyze the unfolded protein response genes, we observed that loss of S1R led to decreased levels of IRE1 and PERK-related effectors and increased over-expression of most of the effectors after a tunicamycin challenge. Finally, S1R deficiency led to alterations in mitochondria bioenergetics with decreased in basal, ATP-linked and non-mitochondrial respiration and following tunicamycin challenge. In conclusion, this new zebrafish model confirmed the importance of S1R activity on ER-mitochondria communication. It will be a useful tool to further analyze the physiopathological roles of S1R.

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The Endogenous Hallucinogen and Trace Amine N,N-Dimethyltryptamine (DMT) Displays Potent Protective Effects against Hypoxia via Sigma-1 Receptor Activation in Human Primary iPSC-Derived Cortical Neurons and Microglia-Like Immune Cells

Frontiers in Neuroscience ◽

10.3389/fnins.2016.00423 ◽

2016 ◽

Vol 10 ◽

Cited By ~ 16

Author(s):

Attila Szabo ◽

Attila Kovacs ◽

Jordi Riba ◽

Srdjan Djurovic ◽

Eva Rajnavolgyi ◽

...

Keyword(s):

Immune Cells ◽

Cortical Neurons ◽

Receptor Activation ◽

Protective Effects ◽

Sigma 1 Receptor ◽

Trace Amine ◽

Sigma 1

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Recent Advances in the Development of Sigma-1 Receptor Ligands

Australian Journal of Chemistry ◽

10.1071/ch14590 ◽

2015 ◽

Vol 68 (4) ◽

pp. 600 ◽

Cited By ~ 6

Author(s):

Madhura Manohar ◽

Samuel D. Banister ◽

Corinne Beinat ◽

James O'Brien-Brown ◽

Michael Kassiou

Keyword(s):

Therapeutic Potential ◽

Molecular Size ◽

Receptor Subtypes ◽

Receptor Ligands ◽

Anatomical Distribution ◽

Sigma 1 Receptor ◽

Affinity Ligands ◽

Selective Ligands ◽

Sigma 1 ◽

Recent Developments

The existence of two distinct sigma (σ) receptor subtypes was established in the early 1990s. Sigma-1 and sigma-2 receptors (S1Rs and S2Rs, respectively) were shown to possess distinct molecular size, anatomical distribution, and ligand discrimination. S2R is overexpressed in numerous human cancers, and has therapeutic potential for the imaging and treatment of certain tumours. In contrast, S1R is more broadly involved in a wide variety of central nervous system (CNS) diseases including motor disorders, memory deficits, depression, schizophrenia, anxiety, pain, drug addiction, and many more. Since the human S1R was cloned in 1996, numerous high affinity ligands with excellent selectivity for S1R have been developed. This review focuses on recent developments in the generation of structurally diverse S1R-selective ligands and novel therapeutic candidates targeting S1Rs.

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Electron Microscopy of a Herpesvirus Isolated from Marek's Disease in Duck and Chicken Embryo Fibroblast Cultures

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100100858 ◽

1968 ◽

Vol 26 ◽

pp. 222-223 ◽

Cited By ~ 1

Author(s):

Keyvan Nazerian

Keyword(s):

Inclusion Bodies ◽

Chicken Embryo Fibroblast ◽

Marek's Disease ◽

Marek’S Disease ◽

Duck Embryo Fibroblast ◽

Multinucleated Cells ◽

Viral Particles ◽

Infected Cells ◽

And Control ◽

Do So

A herpes-like virus has been isolated from duck embryo fibroblast (DEF) cultures inoculated with blood from Marek's disease (MD) infected birds. Cultures which contained this virus produced MD in susceptible chickens while virus negative cultures and control cultures failed to do so. This and other circumstantial evidence including similarities in properties of the virus and the MD agent implicate this virus in the etiology of MD.Histochemical studies demonstrated the presence of DNA-staining intranuclear inclusion bodies in polykarocytes in infected cultures. Distinct nucleo-plasmic aggregates were also seen in sections of similar multinucleated cells examined with the electron microscope. These aggregates are probably the same as the inclusion bodies seen with the light microscope. Naked viral particles were observed in the nucleus of infected cells within or on the edges of the nucleoplasmic aggregates. These particles measured 95-100mμ, in diameter and rarely escaped into the cytoplasm or nuclear vesicles by budding through the nuclear membrane (Fig. 1). The enveloped particles (Fig. 2) formed in this manner measured 150-170mμ in diameter and always had a densely stained nucleoid. The virus in supernatant fluids consisted of naked capsids with 162 hollow, cylindrical capsomeres (Fig. 3). Enveloped particles were not seen in such preparations.

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