Elevated Glucose Levels Favor SARS-CoV-2 Infection and Monocyte Response through a HIF-1α/Glycolysis-Dependent Axis
Authors: Ana Campos Codo
Victor Corasolla Carregari
Rafael Elias Marques
Alexandre Borin
Maria Luiza Moretti
Carlos Alberto Oliveira de Biagi Junior
Eli Mansour
Robson Francisco Carvalho
Gustavo Gastão Davanzo
Karina Bispo Dos Santos
Andre S Vieira
Laís Durço Coimbra
Jeffersson Leandro Jimenez Restrepo
Luciana C Ribeiro
Pedro Henrique Vendramini
Thyago A Nunes
Fabrício Bíscaro Pereira
Vinícius O Boldrini
Guilherme Reis-de-Oliveira
Pedro M Moraes-Vieira
Helison R Carmo
Marcus V Agrela
Alessandro S Farias
Stéfanie Primon Muraro
Juliana Silveira Prodonoff
João Victor Virgilio-da-Silva
Helder I Nakaya
Fernanda Crunfli
Gabriela Fabiano de Souza
Marcelo A Mori
Ana F Bernardes
Licio Augusto Velloso
Andre C Palma
Marco Aurélio Ramirez Vinolo
Raisa G Ulaf
Daniel A Toledo-Teixeira
Lauar de Brito Monteiro
Andrei C Sposito
Pierina Lorencini Parise
André Damasio
José Luiz Proença-Módena
Daniel Martins-de-Souza
Natalia S Brunetti
Matheus Cavalheiro Martini
Journal: Cell Metabolism (2020)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Covid-19; HIF-1alpha; SARS-CoV-2; diabetes; glycolysis
Affiliations: Laboratory of Immunometabolism, Department of Genetics, Evolution, Microbiology and Immunology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil., Department of Genetics, Evolution, Microbiology and Immunology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil., Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil., Department of Clinical and Toxicological analyses, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil., Brazilian Biosciences National Laboratory (LNBio), Campinas, São Paulo, Brazil., Department of Clinical Medicine, School of Medical Sciences, University of Campinas, Campinas, São Paulo, Brazil., Department of Animal Biology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil., Department of Internal Medicine, School of Medical Sciences, University of Campinas, Campinas, São Paulo, Brazil., Hematology and Hemotherapy Center University of Campinas, Campinas, São Paulo, Brazil., Department of Internal Medicine, School of Medical Sciences, University of Campinas, Campinas, São Paulo, Brazil; Obesity and Comorbidities Research Center (OCRC), University of Campinas, São Paulo, Brazil., Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil; Experimental Medicine Research Cluster (EMRC), University of Campinas, São Paulo, Brazil., Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, São Paulo, Brazil., Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil; Obesity and Comorbidities Research Center (OCRC), University of Campinas, São Paulo, Brazil; Experimental Medicine Research Cluster (EMRC), University of Campinas, São Paulo, Brazil., Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil; Experimental Medicine Research Cluster (EMRC), University of Campinas, São Paulo, Brazil; D'Or Institute for Research and Education (IDOR), São Paulo, Brazil; Instituto Nacional de Biomarcadores em Neuropsiquiatria, Conselho Nacional de Desenvolvimento Científico e Tecnológico, São Paulo, Brazil., Department of Genetics, Evolution, Microbiology and Immunology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil; Experimental Medicine Research Cluster (EMRC), University of Campinas, São Paulo, Brazil., Laboratory of Immunometabolism, Department of Genetics, Evolution, Microbiology and Immunology, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil; Obesity and Comorbidities Research Center (OCRC), University of Campinas, São Paulo, Brazil; Experimental Medicine Research Cluster (EMRC), University of Campinas, São Paulo, Brazil. Electronic address: .

Abstract:  COVID-19 can result in severe lung injury. It remained to be determined why diabetic individuals with uncontrolled glucose levels are more prone to develop the severe form of COVID-19. The molecular mechanism underlying SARS-CoV-2 infection and what determines the onset of the cytokine storm found in severe COVID-19 patients are unknown. Monocytes and macrophages are the most enriched immune cell types in the lungs of COVID-19 patients and appear to have a central role in the pathogenicity of the disease. These cells adapt their metabolism upon infection and become highly glycolytic, which facilitates SARS-CoV-2 replication. The infection triggers mitochondrial ROS production, which induces stabilization of hypoxia-inducible factor-1α (HIF-1α) and consequently promotes glycolysis. HIF-1α-induced changes in monocyte metabolism by SARS-CoV-2 infection directly inhibit T cell response and reduce epithelial cell survival. Targeting HIF-1ɑ may have great therapeutic potential for the development of novel drugs to treat COVID-19. 

Retracted: No

PubPeer comments:
 Figure 4C - More similar than expected ![file] , 
 Thanks for the bring my attention of this matter and appreciate comment. We reviewed the figure and found that this was a copy paste error that happened during compiling for publication. We are currently compiling the data to send to the Editor for correction. We apologize for the mistake. , 
 Figure 1D and 1E might be duplicated too. Could authors check please? ![file] , 
 Author could please mention how many times the viral load was performed in Fig 1D, and G. It is surprising to see that the SD is very very narrow and the VL linear to glucose. Also I guess the RNA copy number was calculated in the cells not the supernatant. Is it true VL or ddCt or relative VL? Is it productive replication or just infection? Also it s quite surprising to see that linear VL while other paper failed to show infection in the monocytes. A better description will help other to replicate the finding, particularly the monocyte infection. ![](http: )![file] Ref: https: wwwnc.cdc.gov eid article 26 9 20-1495_article#tnF3 Thank you in advance. , 
 #4 Dear, In both experiments, we have done two independent experiments in triplicate. At the end of the protocol, we removed the supernatant and added 300 uL of TRIzol reagent (Sigma T9424) according to the manufactureru2019s instructions. We performed cDNA conversion using 2000 ug of RNA and with the cDNA, we performed a standard curve using serial dilutions of SARS-CoV-2 cDNA. We then used the standard curve to calculate the number of copies in the volume of the run (in our case 6 uL), normalizing that per mL, as showed in the entire work. We believe that the infection generates productive replication because, in a time course, the number of viral copies increases with time (data not showed), however, we need to perform more experiments to validate that. About the last question, we tested the infection in mouse bone marrow-derived macrophages, peripheral blood monocytes, and peripheral blood macrophages (7 days of M-CSF stimulation). We were not able to infect mouse cells but successfully infected human monocytes and macrophages. We believe the protocol is clear in the star methods section, however, if you have any questions remaining, please contact us. Last, about the paper mentioned, one important difference is the MOI used (0.01 there and 0.1 ours), which in our case is 10 times higher. We also found the paper attached that were able to infect monocytes in vitro (MOI 1) and found circuiting monocytes containing SARS-CoV-2. https: www.biorxiv.org content 10.1101 2020.07.28.225912v1.full.pdf Best regards Gustavo Gastu00e3o Davanzo ,