Bando, Silvia Yumi

Lauterbach, Gerhard da Paz

Moreira-Filho, Carlos Alberto

Vieira, Sandra Elisabete

English

Cadernos Espinosanos (E-Journal)

Human Leukocyte Transcriptional Response to SARS-CoV-2 InfectionSandra Elisabete Vieira0000-0000-0000-0000 ,I Silvia Yumi Bando0000-0000-0000-0000 ,I Gerhard da Paz Lauterbach0000-0000-0000-0000 ,II Carlos AlbertoMoreira-Filho0000-0000-0000-0000 I,*IDepartamento de Pediatria, Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, BR. IIDivisao de Clinica Medica, Hospital das ClinicasHCFMUSP, Sao Paulo, SP, BRVieira SE, Bando SY, Lauterbach GP, Moreira-Filho CA. Human Leukocyte Transcriptional Response to SARS-CoV-2 Infection. Clinics. 2020;75:e2078*Corresponding author. E-mail: carlos.moreira@hc.fm.usp.brIt is necessary to elucidate why in the present coronavirusdisease-19 (COVID-19) global pandemic infection by SARS-CoV-2 affects children, adults, and elderly people differently.Epidemiological and clinical data have shown that mostchildren who test positive for SARS-CoV-2 are either asymp-tomatic or develop mild symptoms (1-3). Several hypotheseshave been suggested to explain these findings: differencesin the immune system of adults and children (and betweenvery young children, pre-schooling children, and adolescents);competition with other respiratory viruses in the respiratorymucosa and lungs influencing the viral load; and differencesbetween adults and children in the expression of the ACE2protein, the SARS-CoV-2 receptor (1). It was recently descri-bed that the hypomethylation and subsequent hyperexpres-sion of the ACE2 gene increased susceptibility for developingCOVID-19 (4).In severe COVID-19 cases, the viral and pulmonary phasesare followed by a final hyperinflammatory phase, which canlead to severe acute respiratory distress syndrome (ARDS),often with a fatal outcome. Here, it should be noted thatchildren are quite susceptible to H1N1-related ARDS (5) andthat the 2003 coronavirus SARS pandemic affected patientsof all ages (6). Therefore, despite the relatively small numberof reported COVID-19 cases in children and the scarceinformation on these cases, it is not possible to assume thatall pediatric COVID-19 cases will follow a mild course (7).The study of the differences between children and adultswith COVID-19 regarding the immune response and diseasecourse represents a unique opportunity for developing newtherapies (8), which is demanded (9) to avoid the collapse ofhealth systems now and in the immediate post-pandemicperiod (2022-2024), as shown by recent epidemiologicalstudies (10). Consequently, we decided to investigate thegenomic basis of those differences through a comparativestudy of the transcriptional responses of human leukocytesto SARS-CoV-2 infection in children and adults, also focusingon the differences between oligosymptomatic and severecases, as further described in the following paragraphs.Severe COVID-19 cases are characterized by a ‘‘cytokinestorm’’ (hypercytokinemia) that promotes hyperinflamma-tion and ARDS (11,12), which is not observed in oligosymp-tomatic cases (13). The inflammatory responses in adults andchildren vary with age, with a progressive increase in inflam-matory cytokines and neutrophil activity, which correlateswith the augmented severity of ARDS in elderly people.Even in pediatric septic shock, the vast majority of geneswith altered expression profiles are in neutrophils (69%) andmonocytes (28%), and just a small minority are in lympho-cytes (14). Therefore, it is quite probable that in circulatingleukocytes distinct transcriptional modules (see below) areassociated with different responses to SARS-CoV-2 inCOVID-19 patients, thus allowing us to delineate adult andchild responses and, in these two groups, the oligosympto-matic and severe case subgroups. The functional analysis ofthese transcriptional modules will allow, as commented onbelow, a better understanding of the pathogenic mechanismstriggered by SARS-CoV-2 and eventually, the identificationof new therapeutic targets.The availability of platforms for large scale gene expres-sion analysis—mainly DNAmicroarrays and next generationsequencing (NGS)—has made it possible for immune res-ponse studies to migrate from a reductionist approach to oneof systems biology (15), enabling a global perception of themolecular, cellular, and tissue events involved in the differenttypes of immune response (16). Studies at this new globaltranscriptome scale have permitted, for instance, a betterunderstanding of the innate and adaptative immune respon-ses, of the defense mechanisms against different pathogens,and the evaluation of the responses to vaccination (17-21).An initial major hurdle in global transcriptome studies washow to analyze and interpret the enormously large geneexpression datasets obtained through DNA microarrays orNGS platforms. The development of statistical and computa-tional tools for the analysis of gene co-expression networkshelped to overcome this limitation (22,23). These tools arepresently used for associating genes and gene expressionprofiles with biological processes and for finding poten-tial therapeutic targets (24-25). Clustering techniques havebeen employed to find genes with similar expression pat-terns in multiple samples, thus identifying modules (26,27).DOI: 10.6061/clinics/2020/e2078Copyright & 2020 CLINICS – This is an Open Access article distributed under theterms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in anymedium or format, provided the original work is properly cited.No potential conflict of interest was reported.Received for publication on May 28, 2020. Accepted for publicationon June 2, 20201EDITORIALTranscriptional modules often represent biological processesand can be phenotype specific (25). The functional enrich-ment among the genes within a module is widely used fordisclosing its biological meaning (25). Moreover, it wasfound that in gene co-expression networks, the highly con-nected genes hold the whole transcriptional network togetherand are either associated with specific cellular processes orlink different biological processes (23). Connectivity measuresare currently used for the hierarchical categorization of genesin transcriptional modules highly correlated with at least onetrait of interest (gender, age, disease features, etc.), helping tofind genes that are highly significant for a certain trait or thatlink molecular pathways in a cell (25).The development of mathematical and computationalmethods for analyzing modular transcriptional repertoireshas been essential for unraveling the human immune defensemechanisms associated with good and bad responses torespiratory viruses (17,28-30). Our group, at the Departmentof Pediatrics, FMUSP, has tackled this approach for inves-tigating the genomic mechanisms associated with the deve-lopment, maturation, and decline of the immune system inhealth and disease (31-33). Recently, studying children undersix months of age hospitalized with acute viral bronchiolitis,we were able to show that in peripheral blood mononuclearcells (PBMC) there are distinct transcriptional modules asso-ciated either with responses to syncytial respiratory virus(HRSV) or rhinovirus (HRV) (20). We also identified host-response molecular markers that could be used for etio-pathogenic diagnosis. The finding of distinct transcriptionalprofiles associated with specific host responses to HRSV orHRV may contribute to unraveling the pathogenic mechan-isms triggered by different respiratory viruses that are indis-tinguishable by clinical presentation, paving the way fornew, specific therapeutic strategies.The experimental approach first adopted for studying thePBMC response to HRSV and HRV is now being used in ourlaboratory to identify the transcriptional responses of humanperipheral blood leukocytes to SARS-CoV-2 following res-piratory tract infection. Relevant knowledge on this subjecthas been newly published. Transcriptome characteristics ofthe bronchoalveolar lavage fluid and peripheral PBMC ofCOVID-19 patients revealed distinct host inflammatory cyto-kine profiles and the association between COVID-19 patho-genesis and excessive cytokine release (34). Compared toother respiratory viruses, SARS-CoV-2 drives a lower anti-viral transcriptional response—low IFN-I and IFN-III levelsand elevated chemokine expression—in accordance with thepro-inflammatory disease state associated with COVID-19(35). In a complementary line of work, COVID-19 patientswere compared to recovered and healthy subjects throughhigh dimensional cytometry, and the subsequent integrationof immune and clinical data revealed different immunotypesrelated to poor clinical course versus improving health (36).Returning to our approach, it aims to identify distinctivetranscriptional modules in the response of human leukocytesto SARS-CoV-2 infection in children and adults, and betweenoligosymptomatic and severe cases in both groups. Thetranscriptomic data thus obtained—distinctive transcrip-tional modules and their associated biological functions,highly connected and high significance genes, etc.—will beintegrated with clinical and demographic data in order togain a better understanding of the molecular mechani-sms involved in the immune response to SARS-CoV-2 and,eventually, for identifying host-response predictors andpotential therapeutic targets for drugs and vaccines.’ ACKNOWLEDGMENTSThis work was funded by Fundação de Amparo à Pesquisa do Estado deSão Paulo (FAPESP) grants no. 2015/22308-2 and 2020/06160-3(‘‘Acordos de Cooperação Covid-19’’), and Conselho Nacional deDesenvolvimento Científico e Tecnológico (CNPq) grant no. 307626/2014-8.’ AUTHOR CONTRIBUTIONSAll the authors contributed equally to this study and have read andapproved the final manuscript.’ REFERENCES1. Brodin P. Why is COVID-19 so mild in children? Acta Paediatr. 2020;109(6):1082-3. https://doi.org/10.1111/apa.152712. 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Human Leukocyte Transcriptional Response to SARSCoV-2 Infection

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Human Leukocyte Transcriptional Response to SARSCoV-2 Infection

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