Evidence of mitochondria origin of SARS-CoV-2 double-membrane vesicles: a review.

Coronavirus Disease-19 (COVID-19) pandemic is caused by the coronavirus SARS-CoV-2 that has infected in a year more than 200 million people and has killed almost 4.5 million people worldwide. This infection affects mainly certain groups of people that have high susceptibility to present severe COVID-19 due to comorbidities. Moreover, the long-COVID-19 comprises a series of symptoms that may remain in some patients for months after infection that further compromises health of individuals. Therefore, this pandemic poses a serious emergency worldwide. Thus, since this pandemic is profoundly affecting economic and social life of societies, a deeper understanding of SARS-CoV-2 infection cycle could help to envisage novel therapeutic alternatives that limit or stop COVID-19. Several recent findings have unexpectedly found that mitochondria play a critical role in SARS-CoV-2 cell infection. Indeed, it has been suggested that this organelle could be the origin of its replication niches, the double membrane vesicles (DMV), as its been observed with another virus. In this regard, mitochondria derived vesicles (MDV), involved in mitochondria quality control, were discovered more than 10 years ago and interestingly there is a population characterized by a double membrane. MDV shedding is induced by mitochondrial stress and it has a fast assembly dynamic, reason that perhaps has precluded their identification in electron microscopy or tomography studies. These and other features of MDV together with recent SARS-CoV-2 protein interactome with the host and other findings linking SARS-CoV-2 to mitochondria, support that these vesicles are the precursors of SARS-CoV-2 induced DMV. In this work, the celular, molecular, phenotypical and biochemical evidence that supports this hypothesis is reviewed and integrated into the current model of SARS-CoV-2 cell infection. In this scheme, some relevant questions are raised as pending topics for research that would help in the near future to test this hypothesis. The intention (abstract truncated).

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A molecular pore spans the double membrane of the coronavirus replication organelle

10.1101/2020.06.25.171686 ◽

2020 ◽

Cited By ~ 3

Author(s):

Georg Wolff ◽

Ronald W.A.L. Limpens ◽

Jessika C. Zevenhoven-Dobbe ◽

Ulrike Laugks ◽

Shawn Zheng ◽

...

Keyword(s):

Drug Target ◽

Rna Synthesis ◽

Transmembrane Protein ◽

Membrane Vesicle ◽

Critical Role ◽

Membrane Vesicles ◽

Genome Replication ◽

Double Membrane ◽

The Core ◽

Novel Drug

Coronavirus genome replication is associated with virus-induced cytosolic double-membrane vesicles, which may provide a tailored micro-environment for viral RNA synthesis in the infected cell. However, it is unclear how newly synthesized genomes and mRNAs can travel from these sealed replication compartments to the cytosol to ensure their translation and the assembly of progeny virions. Here, using cellular electron cryo-microscopy, we unveiled a molecular pore complex that spans both membranes of the double-membrane vesicle and would allow export of RNA to the cytosol. A hexameric assembly of a large viral transmembrane protein was found to form the core of the crown-shaped complex. This coronavirus-specific structure likely plays a critical role in coronavirus replication and thus constitutes a novel drug target

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Dysregulation of lipid metabolism and pathological inflammation in patients with COVID-19

Scientific Reports ◽

10.1038/s41598-021-82426-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Marianna Caterino ◽

Monica Gelzo ◽

Stefano Sol ◽

Roberta Fedele ◽

Anna Annunziata ◽

...

Keyword(s):

Virus Entry ◽

Critical Role ◽

Membrane Vesicles ◽

Virus Propagation ◽

Lipidomic Analysis ◽

Key Factor ◽

Patient Profiles ◽

Cellular Pathways ◽

Severity Degree

AbstractIn recent months, Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread throughout the world. COVID-19 patients show mild, moderate or severe symptoms with the latter ones requiring access to specialized intensive care. SARS-CoV-2 infections, pathogenesis and progression have not been clearly elucidated yet, thus forcing the development of many complementary approaches to identify candidate cellular pathways involved in disease progression. Host lipids play a critical role in the virus life, being the double-membrane vesicles a key factor in coronavirus replication. Moreover, lipid biogenesis pathways affect receptor-mediated virus entry at the endosomal cell surface and modulate virus propagation. In this study, targeted lipidomic analysis coupled with proinflammatory cytokines and alarmins measurement were carried out in serum of COVID-19 patients characterized by different severity degree. Serum IL-26, a cytokine involved in IL-17 pathway, TSLP and adiponectin were measured and correlated to lipid COVID-19 patient profiles. These results could be important for the classification of the COVID-19 disease and the identification of therapeutic targets.

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The Dictatorship of Appearance or Simulation in Present-Day "Democracies" against the Background of the Market Ecocide of the Foundations of Human Life on Earth

Level of Life of the Population of Regions of Russia ◽

10.19181/1999-9836-2019-10075 ◽

2019 ◽

Vol 15 (3) ◽

pp. 82-93

Author(s):

Aleksandr Subetto

Keyword(s):

Social Life ◽

Human Life ◽

Environmental Disaster ◽

Environmental Destruction ◽

Global Environmental ◽

The Social ◽

The World ◽

Point Of No Return ◽

Life On Earth ◽

Near Future

It is proved that the current era is characterized by many governments around the world as dictatorship of "appearance" or "simulation" of the most activities transforming politics, even the tragic events like local ecological catastrophes, local wars, "colour revolutions", the elections in a "theatre", "acting", on the background of market ecocide – really accelerating processes of the first phase of a Global Environmental Disaster, which, at the transition "point of no return" in the near future, may turn into a process of irreversible environmental destruction of all mankind. This dictatorship of "appearance" or simulation as a "curtain" market democracy, hiding the capitalism-led, process of dehumanization of man, is an indicator of the inadequacy of states and political "elites" imperative of survival of mankind, as the imperative out of the ecological impasse of history in market-capitalist format. There comes a reckoning for this departure into the " market-capitalist illusion of apparent prosperity. The societies of the world, including Rossiya, have faced a dilemma:either environmental destruction, or the Noosphere Breakthrough, which, in its essence, is a change in the social organization of social life and its reproduction – the transition from the dominance of capitalism and the market to the Noosphere Ecological Spiritual Socialism on the basis of scientific and educational society and the management of socionatural evolution.

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The YAP/TAZ Signaling Pathway in the Tumor Microenvironment and Carcinogenesis: Current Knowledge and Therapeutic Promises

International Journal of Molecular Sciences ◽

10.3390/ijms23010430 ◽

2021 ◽

Vol 23 (1) ◽

pp. 430

Author(s):

Ángel Ortega ◽

Ivana Vera ◽

Maria P. Diaz ◽

Carla Navarro ◽

Milagros Rojas ◽

...

Keyword(s):

Tumor Microenvironment ◽

Signaling Pathway ◽

Growth Regulation ◽

Current Knowledge ◽

Biological Effects ◽

Critical Role ◽

Binding Motif ◽

Hippo Signaling ◽

Tumor Resistance ◽

Therapeutic Alternatives

The yes-associated protein (YAP) and the transcriptional coactivator with PDZ-binding motif (TAZ) are transcriptional coactivators, members of the Hippo signaling pathway, which play a critical role in cell growth regulation, embryonic development, regeneration, proliferation, and cancer origin and progression. The mechanism involves the nuclear binding of the un-phosphorylated YAP/TAZ complex to release the transcriptional enhanced associate domain (TEAD) from its repressors. The active ternary complex is responsible for the aforementioned biological effects. Overexpression of YAP/TAZ has been reported in cancer stem cells and tumor resistance. The resistance involves chemotherapy, targeted therapy, and immunotherapy. This review provides an overview of YAP/TAZ pathways’ role in carcinogenesis and tumor microenvironment. Potential therapeutic alternatives are also discussed.

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Myocardial Damage by SARS-CoV-2: Emerging Mechanisms and Therapies

Viruses ◽

10.3390/v13091880 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1880

Author(s):

Huyen Tran Ho ◽

Stefan Peischard ◽

Nathalie Strutz-Seebohm ◽

Karin Klingel ◽

Guiscard Seebohm

Keyword(s):

Immune Response ◽

Cell Death ◽

Energy Metabolism ◽

Programmed Cell Death ◽

Viral Replication ◽

Ion Homeostasis ◽

Membrane Vesicles ◽

Cardiac Cells ◽

The Body ◽

Double Membrane

Evidence is emerging that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can infect various organs of the body, including cardiomyocytes and cardiac endothelial cells in the heart. This review focuses on the effects of SARS-CoV-2 in the heart after direct infection that can lead to myocarditis and an outline of potential treatment options. The main points are: (1) Viral entry: SARS-CoV-2 uses specific receptors and proteases for docking and priming in cardiac cells. Thus, different receptors or protease inhibitors might be effective in SARS-CoV-2-infected cardiac cells. (2) Viral replication: SARS-CoV-2 uses RNA-dependent RNA polymerase for replication. Drugs acting against ssRNA(+) viral replication for cardiac cells can be effective. (3) Autophagy and double-membrane vesicles: SARS-CoV-2 manipulates autophagy to inhibit viral clearance and promote SARS-CoV-2 replication by creating double-membrane vesicles as replication sites. (4) Immune response: Host immune response is manipulated to evade host cell attacks against SARS-CoV-2 and increased inflammation by dysregulating immune cells. Efficiency of immunosuppressive therapy must be elucidated. (5) Programmed cell death: SARS-CoV-2 inhibits programmed cell death in early stages and induces apoptosis, necroptosis, and pyroptosis in later stages. (6) Energy metabolism: SARS-CoV-2 infection leads to disturbed energy metabolism that in turn leads to a decrease in ATP production and ROS production. (7) Viroporins: SARS-CoV-2 creates viroporins that lead to an imbalance of ion homeostasis. This causes apoptosis, altered action potential, and arrhythmia.

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The Origin, Dynamic Morphology, and PI4P-Independent Formation of Encephalomyocarditis Virus Replication Organelles

mBio ◽

10.1128/mbio.00420-18 ◽

2018 ◽

Vol 9 (2) ◽

pp. e00420-18 ◽

Cited By ~ 11

Author(s):

C. E. Melia ◽

H. M. van der Schaar ◽

A. W. M. de Jong ◽

H. R. Lyoo ◽

E. J. Snijder ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Rna Synthesis ◽

Electron Tomography ◽

Membrane Vesicles ◽

Encephalomyocarditis Virus ◽

Escape Mutant ◽

Membrane Structures ◽

Double Membrane ◽

Single Membrane ◽

Phosphatidylinositol 4

ABSTRACTPicornaviruses induce dramatic rearrangements of endomembranes in the cells that they infect to produce dedicated platforms for viral replication. These structures, termed replication organelles (ROs), have been well characterized for theEnterovirusgenus of thePicornaviridae. However, it is unknown whether the diverse RO morphologies associated with enterovirus infection are conserved among other picornaviruses. Here, we use serial electron tomography at different stages of infection to assess the three-dimensional architecture of ROs induced by encephalomyocarditis virus (EMCV), a member of theCardiovirusgenus of the family of picornaviruses that is distantly related. Ultrastructural analyses revealed connections between early single-membrane EMCV ROs and the endoplasmic reticulum (ER), establishing the ER as a likely donor organelle for their formation. These early single-membrane ROs appear to transform into double-membrane vesicles (DMVs) as infection progresses. Both single- and double-membrane structures were found to support viral RNA synthesis, and progeny viruses accumulated in close proximity, suggesting a spatial association between RNA synthesis and virus assembly. Further, we explored the role of phosphatidylinositol 4-phosphate (PI4P), a critical host factor for both enterovirus and cardiovirus replication that has been recently found to expedite enterovirus RO formation rather than being strictly required. By exploiting an EMCV escape mutant, we found that low-PI4P conditions could also be overcome for the formation of cardiovirus ROs. Collectively, our data show that despite differences in the membrane source, there are striking similarities in the biogenesis, morphology, and transformation of cardiovirus and enterovirus ROs, which may well extend to other picornaviruses.IMPORTANCELike all positive-sense RNA viruses, picornaviruses induce the rearrangement of host cell membranes to form unique structures, or replication organelles (ROs), that support viral RNA synthesis. Here, we investigate the architecture and biogenesis of cardiovirus ROs and compare them with those induced by enteroviruses, members of the well-characterized picornavirus genusEnterovirus. The origins and dynamic morphologies of cardiovirus ROs are revealed using electron tomography, which points to the endoplasmic reticulum as the donor organelle usurped to produce single-membrane tubules and vesicles that transform into double-membrane vesicles. We show that PI4P, a critical lipid for cardiovirus and enterovirus replication, is not strictly required for the formation of cardiovirus ROs, as functional ROs with typical morphologies are formed under phosphatidylinositol 4-kinase type III alpha (PI4KA) inhibition in cells infected with an escape mutant. Our data show that the transformation from single-membrane structures to double-membrane vesicles is a conserved feature of cardiovirus and enterovirus infections that likely extends to other picornavirus genera.

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Ultrastructural Evidence for Direct Renal Infection with SARS-CoV-2

Journal of the American Society of Nephrology ◽

10.1681/asn.2020040432 ◽

2020 ◽

Vol 31 (8) ◽

pp. 1683-1687 ◽

Cited By ~ 84

Author(s):

Evan A. Farkash ◽

Allecia M. Wilson ◽

Jeffrey M. Jentzen

Keyword(s):

Renal Failure ◽

Light Microscopy ◽

Virus Assembly ◽

Light And Electron Microscopy ◽

Membrane Vesicles ◽

Renal Tissue ◽

Double Membrane ◽

Renal Tubular Epithelium ◽

Renal Tubular ◽

Renal Infection

BackgroundA significant fraction of patients with coronavirus disease 2019 (COVID-19) display abnormalities in renal function. Retrospective studies of patients hospitalized with COVID-19 in Wuhan, China, report an incidence of 3%–7% progressing to ARF, a marker of poor prognosis. The cause of the renal failure in COVID-19 is unknown, but one hypothesized mechanism is direct renal infection by the causative virus, SARS-CoV-2.MethodsWe performed an autopsy on a single patient who died of COVID-19 after open repair of an aortic dissection, complicated by hypoxic respiratory failure and oliguric renal failure. We used light and electron microscopy to examine renal tissue for evidence of SARS-CoV-2 within renal cells.ResultsLight microscopy of proximal tubules showed geographic isometric vacuolization, corresponding to a focus of tubules with abundant intracellular viral arrays. Individual viruses averaged 76 µm in diameter and had an envelope studded with crown-like, electron-dense spikes. Vacuoles contained double-membrane vesicles suggestive of partially assembled virus.ConclusionsThe presence of viral particles in the renal tubular epithelium that were morphologically identical to SARS-CoV-2, and with viral arrays and other features of virus assembly, provide evidence of a productive direct infection of the kidney by SARS-CoV-2. This finding offers confirmatory evidence that direct renal infection occurs in the setting of AKI in COVID-19. However, the frequency and clinical significance of direct infection in COVID-19 is unclear. Tubular isometric vacuolization observed with light microscopy, which correlates with double-membrane vesicles containing vacuoles observed with electronic microscopy, may be a useful histologic marker for active SARS-CoV-2 infection in kidney biopsy or autopsy specimens.

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Examining the Effects of Overtime Work on Subjective Social Status and Social Inclusion in the Chinese Context

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph17093265 ◽

2020 ◽

Vol 17 (9) ◽

pp. 3265 ◽

Cited By ~ 3

Author(s):

Yashuo Chen ◽

Pengbo Li ◽

Chunjiang Yang

Keyword(s):

Social Status ◽

Social Life ◽

Social Inclusion ◽

Relative Deprivation ◽

Critical Role ◽

Negative Relationship ◽

Subjective Social Status ◽

Low Level ◽

Overtime Work ◽

High Level

Although researchers have argued that long work hours have been shown to threaten individual health, lead to work-family conflict, and reduce job performance, the effect of overtime work on social-related outcomes has received little attention. Based on the framework of relative deprivation, we attempt to address this important issue by exploring whether, why, and when individuals’ overtime work influences their social attitudes. By using the data of 400 Chinese employees from the China Labor-Force Dynamics Survey (CLD), we found that overtime work was associated with a low level of subjective social status and social inclusion. In addition, we found that the time type of overtime work (work overtime on weekdays or on weekends and holidays) has a moderating effect on the relationship between overtime work and social inclusion. That is, employees who work overtime on weekdays are unlikely to have a sense of social inclusion. Furthermore, the negative relationship between overtime work and subjective social status was stronger at a low level of fairness rather than a high level of fairness. In contrast, the negative relationship between overtime work and social inclusion was stronger at a high level of fairness rather than a low level of fairness. These findings highlight the critical role of overtime work in social life and also provide novel insights into social intervention aimed at the happiness and harmony of a society.

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Direct lysosome-based autophagy of lipid droplets in hepatocytes

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2011442117 ◽

2020 ◽

Vol 117 (51) ◽

pp. 32443-32452

Author(s):

Ryan J. Schulze ◽

Eugene W. Krueger ◽

Shaun G. Weller ◽

Katherine M. Johnson ◽

Carol A. Casey ◽

...

Keyword(s):

Mammalian Cells ◽

Lipid Droplets ◽

Critical Role ◽

Specific Protein ◽

Lipid Homeostasis ◽

Dynamic Interactions ◽

Double Membrane ◽

Live Cell Microscopy ◽

Cell Microscopy

Hepatocytes metabolize energy-rich cytoplasmic lipid droplets (LDs) in the lysosome-directed process of autophagy. An organelle-selective form of this process (macrolipophagy) results in the engulfment of LDs within double-membrane delimited structures (autophagosomes) before lysosomal fusion. Whether this is an exclusive autophagic mechanism used by hepatocytes to catabolize LDs is unclear. It is also unknown whether lysosomes alone might be sufficient to mediate LD turnover in the absence of an autophagosomal intermediate. We performed live-cell microscopy of hepatocytes to monitor the dynamic interactions between lysosomes and LDs in real-time. We additionally used a fluorescent variant of the LD-specific protein (PLIN2) that exhibits altered fluorescence in response to LD interactions with the lysosome. We find that mammalian lysosomes and LDs undergo interactions during which proteins and lipids can be transferred from LDs directly into lysosomes. Electron microscopy (EM) of primary hepatocytes or hepatocyte-derived cell lines supports the existence of these interactions. It reveals a dramatic process whereby the lipid contents of the LD can be “extruded” directly into the lysosomal lumen under nutrient-limited conditions. Significantly, these interactions are not affected by perturbations to crucial components of the canonical macroautophagy machinery and can occur in the absence of double-membrane lipoautophagosomes. These findings implicate the existence of an autophagic mechanism used by mammalian cells for the direct transfer of LD components into the lysosome for breakdown. This process further emphasizes the critical role of lysosomes in hepatic LD catabolism and provides insights into the mechanisms underlying lipid homeostasis in the liver.

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A molecular pore spans the double membrane of the coronavirus replication organelle

Science ◽

10.1126/science.abd3629 ◽

2020 ◽

Vol 369 (6509) ◽

pp. 1395-1398 ◽

Cited By ~ 26

Author(s):

Georg Wolff ◽

Ronald W. A. L. Limpens ◽

Jessika C. Zevenhoven-Dobbe ◽

Ulrike Laugks ◽

Shawn Zheng ◽

...

Keyword(s):

Drug Target ◽

Rna Synthesis ◽

Transmembrane Protein ◽

Membrane Vesicle ◽

Membrane Vesicles ◽

Genome Replication ◽

Messenger Rnas ◽

Double Membrane ◽

The Core ◽

Cryo Electron Microscopy

Coronavirus genome replication is associated with virus-induced cytosolic double-membrane vesicles, which may provide a tailored microenvironment for viral RNA synthesis in the infected cell. However, it is unclear how newly synthesized genomes and messenger RNAs can travel from these sealed replication compartments to the cytosol to ensure their translation and the assembly of progeny virions. In this study, we used cellular cryo–electron microscopy to visualize a molecular pore complex that spans both membranes of the double-membrane vesicle and would allow export of RNA to the cytosol. A hexameric assembly of a large viral transmembrane protein was found to form the core of the crown-shaped complex. This coronavirus-specific structure likely plays a key role in coronavirus replication and thus constitutes a potential drug target.

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