ORIGINAL RESEARCH

published: 03 November 2020

doi: 10.3389/fcvm.2020.582399

Molecular Basis of Cardiac and

Vascular Injuries Associated With
COVID-19
Mahmood Yaseen Hachim 1 , Saba Al Heialy 1,2 , Abiola Senok 1*, Qutayba Hamid 2,3 and
Alawi Alsheikh-Ali 1*
1
College of Medicine, Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai, United Arab Emirates,
2
Meakins-Christie Laboratories, Research Institute of the McGill University Health Center, Montreal, QC, Canada, 3 College of
Medicine, Sharjah Institute for Medical Research, University of Sharjah, Sharjah, United Arab Emirates

Background: Coronavirus disease 2019 (COVID-19) is a viral respiratory illness caused

by the novel coronavirus SARS-CoV-2. The presence of the pre-existing cardiac disease
is associated with an increased likelihood of severe clinical course and mortality
in patients with COVID-19. Besides, current evidence indicates that a significant
number of patients with COVID-19 also exhibit cardiovascular involvement even in
the absence of known cardiac risk factors. Therefore, there is a need to understand
the underlying mechanisms and genetic predispositions that explain cardiovascular
involvement in COVID-19.
Edited by:
Objectives: In silico analysis of publicly available datasets to decipher the molecular
Mireille Ouimet,
University of Ottawa, Canada basis, potential pathways, and the role of the endothelium in the pathogenesis of cardiac
Reviewed by: and vascular injuries in COVID-19.
Giuseppe Danilo Norata,
University of Milan, Italy
Materials and Methods: Consistent significant differentially expressed genes (DEGs)
Wai Ho Tang, shared by endothelium and peripheral immune cells were identified in five microarray
Guangzhou Medical University, China
transcriptomic profiling datasets in patients with venous thromboembolism “VTE,” acute
*Correspondence:
coronary syndrome, heart failure and/or cardiogenic shock (main cardiovascular injuries
Abiola Senok
Abiola.senok@mbru.ac.ae related to COVID-19) compared to healthy controls. The identified genes were further
Alawi Alsheikh-Ali examined in the publicly available transcriptomic dataset for cell/tissue specificity in lung
Alawi.Alsheikhali@mbru.ac.ae
tissue, in different ethnicities and in SARS-CoV-2 infected vs. mock-infected lung tissues
Specialty section: and cardiomyocytes.
This article was submitted to
Results: We identified 36 DEGs in blood and endothelium known to play key roles
Cardiovascular Metabolism,
a section of the journal in endothelium and vascular biology, regulation of cellular response to stress as well
Frontiers in Cardiovascular Medicine as endothelial cell migration. Some of these genes were upregulated significantly in
Received: 11 July 2020 SARS-CoV-2 infected lung tissues. On the other hand, some genes with cardioprotective
Accepted: 18 September 2020
Published: 03 November 2020
functions were downregulated in SARS-CoV-2 infected cardiomyocytes.
Citation: Conclusion: In conclusion, our findings from the analysis of publicly available
Hachim MY, Al Heialy S, Senok A,
transcriptomic datasets identified shared core genes pertinent to cardiac and
Hamid Q and Alsheikh-Ali A (2020)
Molecular Basis of Cardiac and vascular-related injuries and their probable role in genetic susceptibility to cardiovascular
Vascular Injuries Associated With injury in patients with COVID-19.
COVID-19.
Front. Cardiovasc. Med. 7:582399. Keywords: COVID-19, SARS-CoV-2, cardiac and vascular injuries, transcriptomic, DEG (differentially expressed
doi: 10.3389/fcvm.2020.582399 gene) analysis

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Hachim et al. Cardiovascular Injuries in COVID-19

INTRODUCTION a report from China documenting high levels of troponin or

cardiac arrest in up to 12% of patients without prior history of
Coronavirus disease 2019 (COVID-19) is a viral respiratory cardiovascular disease (6).
illness caused by the novel coronavirus SARS-CoV-2. To date (1st The acute cardiovascular syndrome associated with COVID-
September 2020), the number of laboratory-confirmed cases of 19 includes a variety of clinical presentations of acute
COVID-19 has exceeded 25 million globally, with over 800,000 cardiac injury, cardiomyopathy, and hemodynamic instability.
fatalities (1). The clinical spectrum of COVID-19 ranges from Myocardial injury, arrhythmias, cardiac arrests, heart failure, and
asymptomatic infection to mild to moderate disease in the coagulation abnormality were reported in 7–33% of patients with
majority of patients (2, 3). However, some patients exhibit COVID-19 in China (3, 9). The angiotensin-converting enzyme
a more severe clinical course characterized by multisystemic 2 (ACE-2) receptors used for cellular entry by SARS-CoV-2 are
and life-threatening manifestations with pneumonia and acute expressed in the lung as well as in various organs, including
respiratory distress as prominent features (2–4). Patients with the heart and endothelial cells (10–12). Direct SARS-CoV-2
pre-existing cardiac disease, hypertension, diabetes, and obesity infection of the endothelial cells, along with diffuse endothelial
are more likely to have a severe clinical course with a higher inflammation, has been reported (11). The cytokine storm and
risk of mortality (5–7). In a meta-analysis of 8 studies, including profound inflammation seen in patients with severe COVID-19
46,248 patients, cardiovascular disease was the third most are associated with macrophage and endothelial activation and
common comorbidity in patients with COVID-19 (4). Moreover, surges in the levels of interleukin (IL)-1, IL-6, IL-8, and Tumor
there is increasing evidence that a significant number of patients Necrosis Factor-alpha (TNF-α). Emerging data also indicate a
with COVID-19 have cardiovascular involvement, which further hypercoagulable state in a cluster of patients with COVID-
increases the likelihood of mortality (5, 6, 8, 9). Notably, even in 19 with a high incidence of venous thromboembolism (VTE)
the absence of known cardiac risk factors, patients with COVID- despite the use of prophylactic anticoagulants (13). Studies have
19 may have an increased risk of cardiovascular injury with shown that IL-6, one of the significant cytokines described in the

GRAPHICAL ABSTRACT | SARS-CoV-2 can induce cardiovascular injures in COVID-19 patients by manipulating a core set of genes specific to endothelium in the
lungs, heart, and vessels. This can activate pathways for systemic immune-mediated cardiovascular injuries or increase vulnerability to cardiac injury via inhibition of
cardioprotective proteins. Created with BioRender.com.

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Hachim et al. Cardiovascular Injuries in COVID-19

TABLE 1 | List and details of publicly available transcriptomic datasets used in the identification of differentially expressed genes (DEGs) in cardiovascular injuries.

Cardiovascular injuries Title GSE Sample type Patients Controls

Venous thromboembolism Whole blood gene expression profiles GSE19151 Whole blood 70 63
distinguish patients with single vs. recurrent
venous thromboembolism
Whole blood gene expression profiles GSE48000 Whole blood 109 25
distinguish clinical phenotypes of venous
thromboembolism [Set1]
Gene expression profile of endothelial GSE118259 Endothelial cells 8 5
colony-forming cells (ECFCs) isolated from
patients with unprovoked venous
thromboembolism (uVTE)
Acute coronary syndrome Differential gene expression in GSE19339 White blood cells 4 4
thrombus-derived white blood cells of
patients with acute coronary syndrome
Heart failure and/or Expression data from heart failure vs. control GSE9128 Peripheral blood mononuclear cells 24 12
cardiogenic shock peripheral blood mononuclear cells

Total 5 datasets 215 109

cytokine storm, is associated with vascular leakage, activation of endothelium in the pathogenesis of cardiac and vascular injuries
the coagulation cascade, and cardiomyopathy (14, 15). in COVID-19.
One of the proposed mechanisms of cardiovascular injury
in COVID-19 is direct injury to myocardial cells due to viral
invasion of the vascular endothelium and myocardium (16). The MATERIALS AND METHODS
second postulate is the impact of tissue hypoxia, destabilization
of coronary plaque, and micro-thrombogenesis caused by the Identification of Differentially Expressed
systematic inflammation associated with cytokine storm (16). In Genes in Blood Cells Following
addition, the potential role of genetic susceptibility to COVID-19 Cardiovascular Injuries
related cardiac events has recently been highlighted as a possible Datasets
contributor to the high mortality among African American Publicly available transcriptomic datasets were retrieved from
patients with COVID-19 (17). As cardiovascular involvement Gene Expression Omnibus (GEO) (https://www.ncbi.nlm.
in COVID-19 is now recognized as a predictor of mortality, nih.gov/geo/). Microarray gene expression datasets with the
there is a need to understand the underlying mechanisms and word “venous thromboembolism, acute coronary syndrome,
genetic predisposition. arrhythmia, viral myocarditis, heart failure, and/or cardiogenic
Endothelial cells, like other structural cells, when shock” were selected. Then we selected datasets with human
physiologically activated or during injury like the case of patients’ samples that were compared with age-matched
cardiovascular diseases with or without COVID-19, can release healthy controls and where the samples studied were either
increased levels of circulating phospholipid-rich microvesicles whole blood, peripheral blood cells, or endothelium. No
that can affect recipient cells locally or via the systemic datasets of viral myocarditis or cardiogenic shock fulfilled
circulation (18). Such vesicles, called exosomes, may enclose a these inclusion criteria. The five datasets (215 patients and 109
range of parent cell molecules, including nucleic acids (DNA, healthy control) that fulfilled the inclusion criteria are shown
mRNA, microRNA, and lncRNA), proteins, and lipids (19). in Table 1.
Necrotic or apoptotic processes induced during vascular
endothelium damage can lead to the dissemination of such
exosomes such that mRNA detected in the circulation can be DEGs
representative of cells that do not circulate (20). Sampling and We used GEOquery and limma R packages through the GEO2R
molecular analysis of such circulating cells, extracellular vesicles, tool for each dataset (22). We selected the differentially expressed
nucleic acids, which is referred to as liquid biopsy, is emerging as probes, as previously described (23). Briefly, we sorted the genes
a promising approach for research in cardiovascular injuries (19). related to the filtered probes according to the False Discovery
Recently endothelial, granulocyte, and platelet-derived exosomes Rate (FDR) and selected the top 2,000 differentially expressed
were used to discriminate and map coronary atherosclerotic probes with FDR <0.05 from each dataset. The annotated genes
plaque and calcification in asymptomatic patients (21). In line in each dataset were intersected with DEGs from all other
with this paradigm, we carried out in silico analysis of publicly datasets. Enriched Ontology Clustering for the identified genes
available datasets derived from different cell sources to decipher was performed using the Metascape (http://metascape.org/gp/
the molecular basis, potential pathways, and the role of the index.html#/main/step1).

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Hachim et al. Cardiovascular Injuries in COVID-19

Identification of DEGs in Different Identification of DEGs in SARS-CoV-2

Ethnicities Infected Cells and Lungs
In light of the premise for a potential role for genetic The expression of the shortlisted genes was explored in the
susceptibility to cardiovascular injuries associated with dataset (GSE147507), where RNA-sequencing of transformed
COVID-19, we further explored for the expression of the alveolar lung cells (A549) were mock-treated (n = 6) or infected
identified DEGs in the publicly available dataset (GSE17078) with SARS-CoV-2 (USA-WA1/2020) (n = 6) (25). The same
of blood outgrowth endothelial cells from 27 healthy dataset contains uninfected human lung biopsies, one male (age:
subjects of diverse ages and grouped into Caucasian and 72 years), and one female (age: 60 years), which were used as
African Americans. biological replicates and were compared to lung samples derived
from a single deceased male patient with COVID-19 (age: 74
years). The retrieved data were used to identify DEGs between
Virus Perturbations From GEO
infected and uninfected lung samples using BioJupies online tool
In order to explore if the identified genes showed
(https://amp.pharm.mssm.edu/biojupies/). The normalized gene
differential expression during viral infections and to
expression was used further to estimate infiltrating immune cells
identify the viruses that affect their expression, we used
in the lungs.
the “Gene-virus associations by differential expression
of gene following viral infection” database. “https://
amp.pharm.mssm.edu/Harmonizome/dataset/GEO+ Estimation of Infiltrating Immune Cells in
Signatures+of+Differentially+Expressed+Genes+for+Viral+ the Lungs
Infections.” The normalized gene expression was uploaded to CIBERSORT
(https://cibersort.stanford.edu/) to quantify immune cell
fractions from bulk lung tissue gene expression profile (26).
Lung Gene Expression
To identify which lung cells specifically express the genes of
interest at a significantly higher level compared to other cells, Map of Protein Expression Across Human
we explored LungGENS (Lung Gene Expression iN Single-cell), a Tissues
web-based resource for querying lung single-cell gene expression Tissue specificity of the identified genes was investigated using
databases (24). The Human Protein Atlas (https://www.proteinatlas.org/) (27).

FIGURE 1 | Shared DEGs in the blood and endothelium of patients with venous thromboembolism compared to healthy controls. DEGs in the whole blood of patients
with venous thromboembolism (GSE19151 and GSE48000) were intersected with DEGs in endothelial cells of patients with venous thromboembolism (GSE118259).
Created with BioRender.com.

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Hachim et al. Cardiovascular Injuries in COVID-19

TABLE 2 | List of the 36 shared DEGs in the blood and endothelium of patients with venous thromboembolism compared to healthy controls.

Gene symbol Description Biological process (GO)

AFTPH Aftiphilin GO:0015031 protein transport;GO:0015833 peptide transport;GO:0046907 intracellular transport

AHNAK AHNAK nucleoprotein GO:1901385 regulation of voltage-gated calcium channel activity;GO:1901019 regulation of calcium ion
transmembrane transporter activity;GO:1903169 regulation of calcium ion transmembrane transport
AQR Aquarius intron-binding spliceosomal GO:0006283 transcription-coupled nucleotide-excision repair;GO:0006289 nucleotide-excision
factor repair;GO:0000398 mRNA splicing, via spliceosome
CHD9 Chromodomain helicase DNA GO:0032508 DNA duplex unwinding;GO:0032392 DNA geometric change;GO:0071103 DNA conformation
binding protein 9 change
CNPY2 Canopy FGF signaling regulator 2 GO:0045716 positive regulation of low-density lipoprotein particle receptor biosynthetic
process;GO:0045714 regulation of low-density lipoprotein particle receptor biosynthetic
process;GO:0010870 positive regulation of receptor biosynthetic process
DEDD Death effector domain containing GO:0008625 extrinsic apoptotic signaling pathway via death domain receptors;GO:0097191 extrinsic
apoptotic signaling pathway;GO:0007283 spermatogenesis
DERL2 Derlin 2 GO:1904153 negative regulation of retrograde protein transport, ER to cytosol;GO:1904293 negative
regulation of ERAD pathway;GO:1904152 regulation of retrograde protein transport, ER to cytosol
DICER1 Dicer 1, ribonuclease III GO:0032290 peripheral nervous system myelin formation;GO:0014040 positive regulation of Schwann cell
differentiation;GO:0033168 conversion of ds siRNA to ss siRNA involved in RNA interference
ERCC1 ERCC excision repair 1, GO:1905765 negative regulation of protection from non-homologous end joining at telomere;GO:1904431
endonuclease non-catalytic subunit positive regulation of t-circle formation;GO:1905764 regulation of protection from non-homologous end
joining at telomere
ETS1 ETS proto-oncogene 1, transcription GO:0045648 positive regulation of erythrocyte differentiation;GO:1904996 positive regulation of leukocyte
factor adhesion to vascular endothelial cell;GO:0060055 angiogenesis involved in wound healing
FBXO38 F-box protein 38 GO:0002842 positive regulation of T cell mediated immune response to tumor cell;GO:0002840 regulation of
T cell mediated immune response to tumor cell;GO:0002424 T cell mediated immune response to tumor cell
GADD45GIP1 GADD45G interacting protein 1 GO:0070126 mitochondrial translational termination;GO:0071850 mitotic cell cycle arrest;GO:0070125
mitochondrial translational elongation
HIKESHI Heat shock protein nuclear import GO:1900034 regulation of cellular response to heat;GO:0006606 protein import into nucleus;GO:0034605
factor hikeshi cellular response to heat
IFT52 Intraflagellar transport 52 GO:0035720 intraciliary anterograde transport;GO:0035735 intraciliary transport involved in cilium
assembly;GO:0042733 embryonic digit morphogenesis
LGALS8 Galectin 8 GO:1904977 lymphatic endothelial cell migration;GO:0098792 xenophagy;GO:0036303 lymph vessel
morphogenesis
MRPS11 Mitochondrial ribosomal protein S11 GO:0070126 mitochondrial translational termination;GO:0070125 mitochondrial translational
elongation;GO:0042769 DNA damage response, detection of DNA damage
MTF2 Metal response element binding GO:0061086 negative regulation of histone H3-K27 methylation;GO:0061087 positive regulation of histone
transcription factor 2 H3-K27 methylation;GO:0061085 regulation of histone H3-K27 methylation
MYC MYC proto-oncogene, bHLH GO:0090096 positive regulation of metanephric cap mesenchymal cell proliferation;GO:0090095 regulation
transcription factor of metanephric cap mesenchymal cell proliferation;GO:0090094 metanephric cap mesenchymal cell
proliferation involved in metanephros development
NDUFA4 NDUFA4 mitochondrial complex GO:0006123 mitochondrial electron transport, cytochrome c to oxygen;GO:0006120 mitochondrial electron
associated transport, NADH to ubiquinone;GO:0042775 mitochondrial ATP synthesis coupled electron transport
NDUFB7 NADH:ubiquinone oxidoreductase GO:0006120 mitochondrial electron transport, NADH to ubiquinone;GO:0042775 mitochondrial ATP
subunit B7 synthesis coupled electron transport;GO:0042773 ATP synthesis coupled electron transport
OGT O-linked N-acetylglucosamine GO:0061087 positive regulation of histone H3-K27 methylation;GO:0061085 regulation of histone H3-K27
(GlcNAc) transferase methylation;GO:0043982 histone H4-K8 acetylation
OSBPL8 Oxysterol binding protein like 8 GO:0046326 positive regulation of glucose import;GO:0010891 negative regulation of sequestering of
triglyceride;GO:0046628 positive regulation of insulin receptor signaling pathway
PDCD10 Programmed cell death 10 GO:1903588 negative regulation of blood vessel endothelial cell proliferation involved in sprouting
angiogenesis;GO:0036481 intrinsic apoptotic signaling pathway in response to hydrogen
peroxide;GO:0090051 negative regulation of cell migration involved in sprouting angiogenesis
PRMT2 Protein arginine methyltransferase 2 GO:0019919 peptidyl-arginine methylation, to asymmetrical-dimethyl arginine;GO:0035247 peptidyl-arginine
omega-N-methylation;GO:0060765 regulation of androgen receptor signaling pathway
PUS3 Pseudouridine synthase 3 GO:0031119 tRNA pseudouridine synthesis;GO:1990481 mRNA pseudouridine synthesis;GO:0001522
pseudouridine synthesis
RORA RAR related orphan receptor A GO:0021702 cerebellar Purkinje cell differentiation;GO:0072539 T-helper 17 cell differentiation;GO:0021694
cerebellar Purkinje cell layer formation

(Continued)

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TABLE 2 | Continued

Gene symbol Description Biological process (GO)

RPS15A Ribosomal protein S15a GO:0006614 SRP-dependent cotranslational protein targeting to membrane;GO:0006613 cotranslational
protein targeting to membrane;GO:0000184 nuclear-transcribed mRNA catabolic process,
nonsense-mediated decay
RPS29 Ribosomal protein S29 GO:0006614 SRP-dependent cotranslational protein targeting to membrane;GO:0006613 cotranslational
protein targeting to membrane;GO:0000184 nuclear-transcribed mRNA catabolic process,
nonsense-mediated decay
SLC35A2 Solute carrier family 35 member A2 GO:0072334 UDP-galactose transmembrane transport;GO:0090481 pyrimidine nucleotide-sugar
transmembrane transport;GO:0006012 galactose metabolic process
SON SON DNA and RNA binding protein GO:0048024 regulation of mRNA splicing, via spliceosome;GO:0000281 mitotic cytokinesis;GO:0050684
regulation of mRNA processing
SPAG9 Sperm associated antigen 9 GO:0007257 activation of JUN kinase activity;GO:0043507 positive regulation of JUN kinase
activity;GO:0043506 regulation of JUN kinase activity
TRRAP Transformation/transcription domain GO:0043968 histone H2A acetylation;GO:0043967 histone H4 acetylation;GO:1904837 beta-catenin-TCF
associated protein complex assembly
TTC1 Tetratricopeptide repeat domain 1 GO:0006457 protein folding;GO:0009987 cellular process;GO:0008150 biological_process
TXNL1 Thioredoxin like 1 GO:0045454 cell redox homeostasis;GO:0019725 cellular homeostasis;GO:0055114 oxidation-reduction
process
USP33 Ubiquitin specific peptidase 33 GO:0071108 protein K48-linked deubiquitination;GO:0070536 protein K63-linked
deubiquitination;GO:0051298 centrosome duplication
ZDHHC3 Zinc finger DHHC-type GO:1903546 protein localization to photoreceptor outer segment;GO:0097499 protein localization to
palmitoyltransferase 3 non-motile cilium;GO:0018230 peptidyl-L-cysteine S-palmitoylation

A blood cell-type expression (RNA) option was used to examine genes were vital for pathways involved in cell homeostasis,
the cell specificity of the identified genes. Normalized expressions response to stress, and cellular metabolism. These include
(NX) for 18 blood cell types and total peripheral blood pathways related to targets of C-MYC transcriptional activation
mononuclear cells (PBMC) were explored. (MYC, TRRAP, PDCD10, OGT, USP33, ZDHHC3, and ETS1),
regulation of cellular response to stress (ERCC1, MYC,
Identification of Differentially Expressed SPAG9, PDCD10, DERL2, and HIKESHI), and endothelial
Genes in SARS-CoV-2 Infected cell migration (ETS1, LGALS8, and PDCD10). Figure 2
Human-Induced Pluripotent Stem shows the list of biological pathways associated with the
DEGs. Four genes MYC, ETS1, OGT, and PDCD10 were
Cell-Derived Cardiomyocytes shown to be common between the top pathways indicating
The expression of the shortlisted genes was explored in the their significant molecular and biological role:. They are all
transcriptomic dataset “GSE150392” which is derived from enriched in the PID MYC ACTIV PATHWAY, suggesting
human-induced pluripotent stem cell-derived cardiomyocytes that they are targets of C-MYC transcriptional activation.
infected in vitro with SARS-CoV-2. The genes which showed The proto-oncogene c-Myc is vital for vascular development.
significant differential expression between SARS-CoV-2 and Gene expression analysis of c-Myc-deficient endothelial cells
mock-infected cells were identified. showed that the senescent phenotype of c-Myc is needed
for the prevention of vascular pro-inflammatory phenotype
RESULTS (28). Global or endothelial and hematopoietic cell-specific
loss of c-Myc leads to defects in vasculogenesis and primitive
Whole Blood and Endothelium Shared erythropoiesis (29).
DEGs in Patients With Venous
Thromboembolism SON, OGT, and RORA Are Differentially
DEGs in the whole blood of patients with VTE relative to healthy
controls (GSE19151 and GSE48000) were intersected with DEGs Expressed in the Peripheral Blood of
in endothelial cells of patients with VTE relative to healthy Patients With Acute Coronary Syndrome
controls (GSE118259), and 36 genes were identified as DEGs and Heart Failure
common to the three datasets, suggestive of their role in VTE The 36 genes identified to be specific to VTE were intersected
(Figure 1, Table 2). with DEGs in thrombus-derived white blood cells of patients with
acute coronary syndrome vs. controls (GSE19339) and peripheral
The 36 Shared DEGs Play an Essential Role blood mononuclear cells of patients with heart failure vs. control
in Endothelium Biology (GSE9128) (Figure 3). Four genes were shared between VTE
To understand the role of the identified 36 genes, we explored and acute coronary syndrome (MTF2, TXNL1, PRMT2, and
their shared biological pathways and found that several of these ERCC2), and ten genes were shared between VTE and heart

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FIGURE 2 | Top pathways enriched with shared DEGs in the blood and endothelium of patients with VTE compared to healthy controls. Created with BioRender.com.

failure (DICER1, CHD9, MYC, HIKESHI, USP33, AQR, DEDD, SARS-CoV strains from infected lung epithelial cells (SARS-
DERL2, CNPY2, and PUS3). Only three genes, SON (SON DNA CoV, SARS-dORF6, or SARS-BatSRBD “GSE47960, GSE47961
and RNA binding protein), OGT (O-linked N-acetylglucosamine and GSE50000”; icSARS-CoV or the icSARS-dORF6 mutant
[GlcNAc] transferase), and RORA (RAR related orphan receptor “GSE37827”) and a SARS CoV MA15 infection in C57Bl/6
A) were shared by all the three conditions. mouse model “GSE33266.” The genes which showed consistent
differential expression in different datasets in response to SARS-
CoV infections were SON, OGT, PRMT2 (protein arginine
SON, OGT, and RORA Expression in methyltransferase 2), TXNL1 (thioredoxin like 1), CNPY2
Healthy Endothelium of African Americans (canopy FGF signaling regulator 2), MRPS11 (mitochondrial
To explore the premise of genetic susceptibility for COVID-19 ribosomal protein S11), SPAG9 (sperm associated antigen 9),
related cardiac events, we explored the gene expression of the MTF2 (metal response element-binding transcription factor 2),
three shared DEGs (SON, OGT, and RORA) in the publicly CHD9 (chromodomain helicase DNA binding protein 9), and
available dataset (GSE17078) of blood outgrowth endothelial RPS29 (ribosomal protein S29).
cells from 27 healthy Caucasian and African American subjects.
The findings show that SON, OGT, and RORA are significantly
downregulated in the healthy endothelium of African Americans
Lung Single-Cell Expression of DEGs
The cellular composition of the lung is 40–50% endothelial
compared to Caucasians (Figure 4).
cells, which differentiate in parallel with epithelial cells to form
gas exchange units which are in contact with the external
Expression of the DEGs in Viral Infections environment and thus need to ensure a rapid immune response
All 36 DEGs showed differential expression during viral (30). In lung diseases, including infections, the transcriptomes
infections as per the “Gene-virus associations by differential of endothelial cells, pericyte/smooth muscle cells, fibroblasts,
expression of gene following viral infection” database. The most and macrophage clusters showed that endothelial cells had the
frequently identified viruses affecting most of the genes were most differentially expressed gene profile compared to other cell

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Hachim et al. Cardiovascular Injuries in COVID-19

FIGURE 3 | Shared peripheral blood DEGs in patients with venous thromboembolism, acute coronary syndrome, and heart failure. Created with BioRender.com.

FIGURE 4 | The mRNA expression of SON, OGT, and RORA in the publicly available dataset (GSE17078) of blood outgrowth endothelial cells from 27 healthy
subjects of diverse ages and grouped into Caucasian and African Americans. Created with BioRender.com.

types (31). We speculated that if we found common differentially able further to understand the link between COVID-19 and
expressed genes shared between the two cell types and which associated endothelium injuries. Querying lung single-cell gene
could be affected by SARS-CoV-2 infection, then we may be expression databases showed that expression of some of the

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Hachim et al. Cardiovascular Injuries in COVID-19

FIGURE 5 | Expression of DEGs in different lung cell types. Created with BioRender.com.

36 DEGs was significantly higher in lung endothelial cells

(PRMT2, OGT, MTF2, and CHD9), lung fibroblasts (PRMT2,
OGT, MTF2, CHD9, TXNL1, CNPY2, SPAG9, MRPS11, SON,
and RPS29) and epithelial cells (TXNL1, CNPY2, and SPAG9).
The only gene whose expression was found to also be related to
myeloid/immune cells was RPS29. Figure 5 shows the DEGs and
their expression in different cells. Details of peak expressions for
each DEG in different cells is provided in Supplementary Table.

SNPs in the Identified DEGs With

Significant Association to COVID-19
We searched for the COVID-19 GWAS (https://grasp.nhlbi.
nih.gov/Covid19GWASResults.aspx) looking for Annotated top
results (only variants with P<1E-5) in 1,723 positive cases vs.
11,409 negative controls and found that none of the 36 genes
identified carry SNPs with significant association to COVID-
19, indicating that these genes are differentially expressed FIGURE 6 | Identification of differentially expressed genes in SARS-CoV-2
during infection or disease as a dynamic response to stimuli infected cells. The shortlisted genes expression was explored in the dataset
or condition. (GSE147507), where RNA-Sequencing of transformed alveolar lung cells
(A549) were mock-treated (n = 6) or infected with SARS-CoV-2
(USA-WA1/2020) (n = 6). Created with BioRender.com.
RPS29 and SPAG9 in SARS-CoV-2 Infected
Lung Epithelial Cells
The expression of the 36 DEGs was examined in mock vs. SARS-
CoV-2 infected lung epithelial cells. Although most of the genes Cardiac Protective Genes Are
were upregulated by the virus infection, only RPS29 and SPAG9
Downregulated in Human-Induced
showed significant upregulation, as shown in Figure 6.
Pluripotent Stem Cell-Derived
SPAG9 and RPS29 in Immune Cells Cardiomyocytes Infected With
We sought to identify which immune cell expresses the highest SARS-CoV-2
level of RPS29 and SPAG9. Our findings indicate that RPS29 We explored the novel transcriptomic dataset “GSE150392”
showed low cell type specificity but was higher in T cells while which is derived from in vitro work in which human-induced
SPAG9 was enriched specifically in neutrophils and basophils pluripotent stem cell-derived cardiomyocytes were infected
(Figure 7). with SARS-CoV-2. Nine of the 36 DEGs showed significant

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Hachim et al. Cardiovascular Injuries in COVID-19

FIGURE 7 | Immune cells specificity of the identified genes (RPS29 and SPAG9) using The Human Protein Atlas. A blood cell-type expression (RNA) option was used
to examine the cell specificity of the identified genes. Normalized eXpression (NX) for 18 blood cell types and total peripheral blood mononuclear cells (PBMC) were
explored. Created with BioRender.com.

differential expression between SARS-CoV-2 and mock-infected cardiovascular complications seen in COVID-19 patients. To
cells (Table 3). Four genes (NDUFA4, NDUFB7, MRPS11, and achieve that, we started by comparing cases to control in each
HIKESHI) were downregulated by SARS-CoV-2 while the of these diseases to find their shared DEGs, and then determine
remaining five genes (CHD9, MTF2, RORA, MYC, and ETS1) if these genes were also triggered specifically in COVID-19. The
were upregulated. dataset we used for validation was Lung cells infected with SARS-
CoV-2 (which is one of the few datasets available). As these
identified genes were found to be expressed in lung cells, we
DISCUSSION postulate that they might represent the core machinery genes and
Although respiratory failure has been the primary concern in the link between COVID-19, which is, in essence, lung infection
COVID-19 infection, cardiac injury manifested by a rise in high- and cardiovascular injuries, which are systemic consequences.
sensitivity troponin has gained considerable attention due to its While it would be ideal to utilize datasets derived from COVID-
reported association with mortality (3, 5). A higher incidence 19 patients with cardiovascular outcomes for such comparative
of acute onset heart failure, myocardial infarction, myocarditis, analysis, these are currently not available. Nevertheless, the
and cardiac arrest in COVID-19 patients is documented in findings from this study provide important new information that
the literature (9). On the basis of this, we hypothesized that expands our current understanding of cardiovascular injuries
a common molecular pathway shared between these common in COVID-19.
cardiovascular diseases might be activated in SARS-CoV-2 From our comprehensive in silico approach, we identified
infection and thus provide an explanation for the high rate of 36 DEGs in the blood and endothelium of patients with VTE.

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Hachim et al. Cardiovascular Injuries in COVID-19

TABLE 3 | List of genes significantly altered in human-induced pluripotent stem cell-derived cardiomyocytes infected with SARS-CoV-2 in vitro extracted from
(GSE150392) dataset.

Gene logFC AveExpr t P-value Adjusted P-value B

NDUFA4 −1.64768 8.542878 −3.32788 0.010298 0.05 −3.04532

NDUFB7 −1.60553 6.279966 −3.27608 0.011129 0.05 −3.1789
MRPS11 −1.44701 5.208024 −3.69074 0.006038 0.04 −2.54455
HIKESHI −1.4191 5.802794 −3.37874 0.009546 0.05 −3.02343
CHD9 0.805961 6.451166 3.302155 0.010702 0.05 −3.13644
MTF2 0.810151 5.232699 3.511189 0.007846 0.04 −2.81722
RORA 1.288202 3.65585 4.223798 0.002848 0.02 −1.69915
MYC 1.848579 6.041511 6.609779 0.000162 0.01 1.25177
ETS1 1.886356 4.778408 4.258643 0.002716 0.02 −1.70091

FIGURE 8 | Role of RPS29 and SPAG9 genes in SARS-COV-2 related cardiovascular injuries. (1) Lung viral infection (2) upregulates RPS29 that can (3) stimulate
hematopoietic stem cells and red blood cell development to provide immune cells like neutrophils (4) to reach the lung, (5) the virus-induced upregulation of SPAG9
might (6) induce antibodies against it that might cross-(7) react with the heart cytoskeleton and cause cardiac damage in the form of myocarditis and cardiac
dysfunction. Created with BioRender.com.

Among these were genes known to play key roles in endothelium expressed in the peripheral blood of patients with acute coronary
and vascular biology, with several being vital for pathways for C- syndrome and heart failure. These findings implicate SON, OGT,
MYC transcriptional activation, regulation of cellular response to and RORA as shared core genes in cardiac and vascular-related
stress as well as endothelial cell migration. In addition, some of injuries. As these DEGs were also shared with mesenchymal
the genes involved in endothelial cell migration (ETS1, LGALS8, cells of the lung, we speculate that they may represent the
and PDCD10) are also known to be associated with perturbations missing link between lung damage and related cardiovascular
during viral infection (32–37). Notably, of the 36 DEGs injuries reported in patients with COVID-19 patients. SON gene
identified, three genes, namely SON, OGT, and RORA, were also encodes an RNA-binding protein that promotes the splicing

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Hachim et al. Cardiovascular Injuries in COVID-19

of many cell-cycle and DNA-repair transcripts and maintains All the 36 DEGs showed differential expression during viral
accurate splicing for a subset of Human pre-mRNAs (38). SON infections, and the most frequently identified viruses were
is involved in pathways regulating virus infection like influenza SARS-CoV strains. Specifically, in SARS-CoV-2 infected lung
virus infection as its deletion can lead to reduced influenza epithelial cells, RPS29 and SPAG9 genes were significantly
viral RNA levels and decreased viral infection suggesting that upregulated. RPS29, which was the only DEG found to be specific
SON is needed for influenza virus replication (39). In human- to myeloid/immune Cells (S1.21) with TPM of 1,205.92 and
induced pluripotent stem cell-derived multipotent cardiac intermediate fibroblast 2 (S2.5) with TPM of 1,098.99 in the
progenitor cells, knockdown of SON reduced proliferation and lung, encodes for a ribosomal protein with an established role
differentiation of cardiomyocytes, while increasing fibroblasts in hematopoietic stem cells and red blood cell development
(40). OGT is an O-GlcNAc transferase that catalyzes the (48). RPS29 is a component of the small 40S ribosomal subunit
addition of the O-GlcNAc post-translational modification to and needed for rRNA processing and ribosome biogenesis (49).
proteins, which is essential in regulating the stress response, Germ-line mutation in RPS29 cause Diamond-Blackfan anemia,
differentiation, nutrient sensing, and autophagy (41). O-GlcNAc which is an inherited bone marrow failure syndrome (49).
level is increased during ischemia-reperfusion or hemorrhagic RNA-seq analysis of acute myocardial infarction samples has
shock with a cardioprotective effect making augmentation shown that RPS29 was one of the top upregulated genes (50).
of O-GlcNAc levels a potential new therapeutic option for Interestingly, RPS29 has been reported to be upregulated in
cardiovascular dysfunction or ischemia/reperfusion (42). RORA A549 cells infected with the novel H3N2 Swine Influenza virus
is a nuclear receptor retinoic acid-related orphan receptor-α and the 2009 H1N1 pandemic Influenza virus (51). It was
that has been recently identified in the heart to inhibit ANG also upregulated in inflammatory conditions like periodontitis
II-induced pathological hypertrophy and cardiomyocyte death, and associated with raised IFN-α (52). It is likely that the
repress IL-6 transcription, and its level is reduced in failing mouse upregulation of RPS20 in viral infection provides a mechanism
and human hearts (43). RORA deficient staggered mice subjected for stimulation of hematopoietic stem cells and red blood cell
to myocardial ischemia/reperfusion injury show significantly development for increased production of immune cells like
increased myocardial infarct size, myocardial apoptosis, and neutrophils for recruitment to the site of infection. SPAG9
exacerbated contractile dysfunction compared to wild-type is known to induce an immune response and to regulate
mice (44). Moreover, mice with cardiomyocyte-specific RORA JNK and mitogen-activated protein kinases (MAPKs) signaling
overexpression were less vulnerable to injury (44). RORA has pathways, cell cycle progression, and matrix metalloproteinases
been described as a transcription factor which ties metabolic (53). SPAG9 is involved in the trafficking of endocytic vesicles
and inflammatory signaling pathways. In fact, macrophages from within the intercellular bridge (54). SPAG9 antibody in serum
staggerer mice (which have a deletion in RORA) overexpress appears to be related to the type of lung cancer, indicating its
Il1b following LPS stimulation suggesting an anti-inflammatory specificity to lung-related tissues (55). It is one of the cardiac
role for RORA (45). One mechanism that has been postulated cytoskeleton and sarcomere assembly and function genes which
involves the role of RORA in inducing IκBα, which negatively are enhanced in mice with the deleted muscleblind-like family
regulated the NFκB signaling pathway (46). However, it has been of splice regulators involved in cardiac dysfunction (56). The
suggested that RORA may play a dual role in tissue and cell- virus-induced upregulation of SPAG9 might induce antibodies
dependent manner. For example, in adipose tissue RORA may against it that might cross-react with the heart cytoskeleton
play a pro-inflammatory role by driving endoplasmic reticulum and cause cardiac damage in the form of myocarditis and
stress (47). Interestingly in human-induced pluripotent stem, cardiac dysfunction. In Figure 8, we illustrate the pathway for
cell-derived cardiomyocytes infected in vitro with SARS-CoV-2, the postulated role of RPS29 and SPAG9 genes in SARS-COV-2
the expression of RORA was upregulated, and we speculate related cardiovascular injuries.
that this might be a cardioprotective response to direct Our analysis of the SARS-CoV-2 infected cardiomyocyte
viral invasion. Furthermore, SON, OGT, and RORA regulate derived dataset showed that of the 36 DEGs identified in
the maintenance and differentiation of stem cells, including this study, four genes (NDUFA4L2; NDUFB7; MRPS11;
endothelial progenitor cells (EPCs) (6). They are preferentially HIKESHI) which are known to be cardioprotective were
expressed in undifferentiated stem cells but downregulated downregulated. NDUFA4L2 plays a role in protecting
during stem cell differentiation (7–9). The ability of vascular cardiomyocytes from apoptosis and mitochondrial dysfunction
endothelial cells to repair relies on the EPCs (11). As the during ischemia/reperfusion event, while NDUFB7 has been
occurrence of cardiovascular events during COVID-19 suggests linked with mitochondrial dysfunction and cardiomyocyte
that targeting the endothelium is part of the viral infection senescence (57, 58). MRPS11 is a mitochondrial gene involved in
course, we surmise COVID-19 patients who have a pre-existing sex-specific cardiac structure and function alterations (59). Heat
genetic propensity for low SON, OGT, and RORA expression may shock proteins are involved in protecting the heart against heart
therefore be more susceptible to cardiac damage. Our findings failure by facilitating the removal of misfolded and degraded
of significant downregulation of SON, OGT, and RORA in proteins (60), and HIKESHI plays a role in heat-shock stress
healthy endothelium of African Americans is consistent with this response regulation to protect cells from heat shock damages.
hypothesis. This may explain the increased risk of cardiovascular This finding suggests that in addition to the proposed RPS29
injury among African American patients with COVID-19. and SPAG9 induced cardiac damage pathway alluded to earlier,

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Hachim et al. Cardiovascular Injuries in COVID-19

SARS-CoV-2 also employs a mechanism of downregulation of with patients with COVID−19 infection vs. those without
cardioprotective genes to promote cardiac injury. COVID-19 and a similar CVR outcome will be useful in
In conclusion, our findings from the analysis of publicly pinpointing specific genes.
available transcriptomic datasets identified three shared core
genes pertinent to cardiac and vascular-related injuries. The DATA AVAILABILITY STATEMENT
possibility for their role in genetic susceptibility to cardiovascular
injury in patients with COVID-19 was highlighted. In addition, The raw data supporting the conclusions of this article will be
it is likely that a combination of RPS29 and SPAG9 genes made available by the authors, without undue reservation.
induced pathways, as well as downregulation of cardioprotective
genes, contribute to cardiac and vascular events in patients AUTHOR CONTRIBUTIONS
with COVID-19.
Given that our analysis is in silico, experimental validation All authors listed have made a substantial, direct and
of our findings suggesting the potential role in genetic intellectual contribution to the work, and approved it
susceptibility such as in vitro experiments on endothelial cells for publication.
exposed to SARS-CoV-2 antigens are needed to enable a
better understanding of cardiovascular events associated with SUPPLEMENTARY MATERIAL
SARS-CoV-2 infection. The main limitation here is that the study
is performed on the premise that venous thromboembolism, The Supplementary Material for this article can be found
acute coronary syndrome, and heart failure might be common online at: https://www.frontiersin.org/articles/10.3389/fcvm.
during COVID-19 infection. However, in silico analysis of studies 2020.582399/full#supplementary-material

REFERENCES 12. Hamming I, Timens W, Bulthuis ML, Lely AT, Navis G, van Goor H. Tissue
distribution of ACE2 protein, the functional receptor for SARS coronavirus.
1. Hopkins J. Coronavirus resource center John Hopkins University of medicine. A first step in understanding SARS pathogenesis. J Pathol. (2004) 203:631–
In: COVID-19 Dashboard by the Center for Systems Science and Engineering 7. doi: 10.1002/path.1570
(CSSE) at Johns Hopkins University (JHU). ArcGIS; Johns Hopkins University 13. Klok FA, Kruip M, van der Meer NJM, Arbous MS, Gommers
(2020). D, Kant KM, et al. Incidence of thrombotic complications in
2. Chen N, Zhou M, Dong X, Qu J, Gong F, Han Y, et al. Epidemiological critically ill ICU patients with COVID-19. Thromb Res. (2020)
and clinical characteristics of 99 cases of 2019 novel coronavirus 191:145–7. doi: 10.1016/j.thromres.2020.04.013
pneumonia in Wuhan, China: a descriptive study. Lancet. (2020) 14. Kanda T, Takahashi T. Interleukin-6 and cardiovascular diseases. Japan Heart
395:507–13. doi: 10.1016/S0140-6736(20)30211-7 J. (2004) 45:183–93. doi: 10.1536/jhj.45.183
3. Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, et al. Clinical 15. Levi M, Keller TT, van Gorp E, ten Cate H. Infection and
features of patients infected with 2019 novel coronavirus in Wuhan, inflammation and the coagulation system. Cardiovasc Res. (2003)
China. Lancet. (2020) 395:497–506. doi: 10.1016/S0140-6736(20) 60:26–39. doi: 10.1016/S0008-6363(02)00857-X
30183-5 16. Clerkin KJ, Fried JA, Raikhelkar J, Sayer G, Griffin JM, Masoumi A,
4. Yang J, Zheng Y, Gou X, Pu K, Chen Z, Guo Q, et al. Prevalence et al. COVID-19 and cardiovascular disease. Circulation. (2020) 141:1648–
of comorbidities and its effects in patients infected with SARS-CoV-2: 55. doi: 10.1161/CIRCULATIONAHA.120.046941
a systematic review and meta-analysis. Int J Infect Dis. (2020) 94:91– 17. Giudicessi JR, Roden DM, Wilde AAM, Ackerman MJ. Genetic
5. doi: 10.1016/j.ijid.2020.03.017 susceptibility for COVID-19-associated sudden cardiac death in African
5. Wang D, Hu B, Hu C, Zhu F, Liu X, Zhang J, et al. Clinical characteristics of Americans. Heart rhythm. (2020) 17:1487–92. doi: 10.1016/j.hrthm.2020.
138 hospitalized patients with 2019. Novel coronavirus-infected pneumonia 04.045
in Wuhan, China. JAMA. (2020) 323:1061–9. doi: 10.1001/jama.2020.1585 18. Badimon L, Suades R, Vilella-Figuerola A, Crespo J, Vilahur G, Escate R,
6. Zheng YY, Ma YT, Zhang JY, Xie X. COVID-19 and the cardiovascular system. et al. Liquid biopsies: microvesicles in cardiovascular disease. Antioxid Redox
Nat Rev Cardiol. (2020) 17:259–60. doi: 10.1038/s41569-020-0360-5 Signal. (2019) 33:645–62. doi: 10.1089/ars.2019.7922
7. Zhou F, Yu T, Du R, Fan G, Liu Y, Liu Z, et al. Clinical course 19. Zhou B, Xu K, Zheng X, Chen T, Wang J, Song Y, et al. Application of
and risk factors for mortality of adult inpatients with COVID-19 in exosomes as liquid biopsy in clinical diagnosis. Signal Transduct Target Ther.
Wuhan, China: a retrospective cohort study. Lancet. (2020) 395:1054– (2020) 5:144. doi: 10.1038/s41392-020-00258-9
62. doi: 10.1016/S0140-6736(20)30566-3 20. Carrió I, Flotats A. Liquid biopsies and molecular imaging: friends or foes?
8. Kwenandar F, Japar KV, Damay V, Hariyanto TI, Tanaka M, Lugito NPH, et al. Clin Transl Imaging. (2020) 8:47–50. doi: 10.1007/s40336-019-00350-3
Coronavirus disease 2019 and cardiovascular system: a narrative review. Int J 21. Chiva-Blanch G, Padró T, Alonso R, Crespo J, Perez de Isla L, Mata
Cardiol Heart Vasc. (2020) 29:100557. doi: 10.1016/j.ijcha.2020.100557 P, et al. Liquid biopsy of extracellular microvesicles maps coronary
9. Shi S, Qin M, Shen B, Cai Y, Liu T, Yang F, et al. Association of cardiac injury calcification and atherosclerotic plaque in asymptomatic patients with
with mortality in hospitalized patients with COVID-19 in Wuhan, China. familial hypercholesterolemia. Arterioscler Thromb Vasc Biol. (2019) 39:945–
JAMA Cardiol. (2020) 5:802–10. doi: 10.1001/jamacardio.2020.0950 55. doi: 10.1161/ATVBAHA.118.312414
10. Ferrario CM, Jessup J, Chappell MC, Averill DB, Brosnihan KB, Tallant EA, 22. Barrett T, Wilhite SE, Ledoux P, Evangelista C, Kim IF, Tomashevsky M, et al.
et al. Effect of angiotensin-converting enzyme inhibition and angiotensin II NCBI GEO: archive for functional genomics data sets—update. Nucleic Acids
receptor blockers on cardiac angiotensin-converting enzyme 2. Circulation. Res. (2012) 41:D991–5. doi: 10.1093/nar/gks1193
(2005) 111:2605–10. doi: 10.1161/CIRCULATIONAHA.104.510461 23. Hachim MY, Al Heialy S, Hachim IY, Halwani R, Senok AC, Maghazachi AA,
11. Varga Z, Flammer AJ, Steiger P, Haberecker M, Andermatt R, Zinkernagel AS, et al. Interferon-Induced Transmembrane Protein (IFITM3) is upregulated
et al. Endothelial cell infection and endotheliitis in COVID-19. Lancet. (2020) explicitly in sars-cov-2 infected lung epithelial cells. Front Immunol. (2020)
395:1417–8. doi: 10.1016/S0140-6736(20)30937-5 11:1372. doi: 10.3389/fimmu.2020.01372

Frontiers in Cardiovascular Medicine | www.frontiersin.org 13 November 2020 | Volume 7 | Article 582399

Hachim et al. Cardiovascular Injuries in COVID-19

24. Du Y, Guo M, Whitsett JA, Xu Y. ’LungGENS’: a web-based tool for mapping 42. Ferron M, Denis M, Persello A, Rathagirishnan R, Lauzier B.
single-cell gene expression in the developing lung. Thorax. (2015) 70:1092– Protein O-GlcNAcylation in cardiac pathologies: past, present,
4. doi: 10.1136/thoraxjnl-2015-207035 future. Front Endocrinol. (2019) 9:819. doi: 10.3389/fendo.2018.
25. Blanco-Melo D, Nilsson-Payant BE, Liu W-C, Møller R, Panis M, Sachs 00819
D, et al. SARS-CoV-2 launches a unique transcriptional signature 43. Beak JY, Kang HS, Huang W, Myers PH, Bowles DE, Jetten AM,
from in vitro, ex vivo, and in vivo systems. bioRxiv [Preprint]. et al. The nuclear receptor RORα protects against angiotensin
(2020). doi: 10.1101/2020.03.24.004655 II-induced cardiac hypertrophy and heart failure. Am J Physiol
26. Chen B, Khodadoust MS, Liu CL, Newman AM, Alizadeh AA. Profiling Heart Circ Physiol. (2019) 316:H186–200. doi: 10.1152/ajpheart.005
tumor infiltrating immune cells with CIBERSORT. Methods Mol Biol. (2018) 31.2018
1711:243–59. doi: 10.1007/978-1-4939-7493-1_12 44. He B, Zhao Y, Xu L, Gao L, Su Y, Lin N, et al. The nuclear
27. Uhlén M, Fagerberg L, Hallström BM, Lindskog C, Oksvold P, Mardinoglu melatonin receptor RORα is a novel endogenous defender against
A, et al. Tissue-based map of the human proteome. Science. (2015) myocardial ischemia/reperfusion injury. J Pineal Res. (2016) 60:313–
347:1260419. doi: 10.1126/science.1260419 26. doi: 10.1111/jpi.12312
28. Florea V, Bhagavatula N, Simovic G, Macedo FY, Fock RA, Rodrigues CO. 45. Nejati Moharrami N, Bjørkøy Tande E, Ryan L, Espevik T, Boyartchuk V.
c-Myc is essential to prevent endothelial pro-inflammatory senescent RORα controls inflammatory state of human macrophages. PLoS ONE. (2018)
phenotype. PLoS ONE. (2013) 8:e73146. doi: 10.1371/journal.pone. 13:e0207374. doi: 10.1371/journal.pone.0207374
0073146 46. Delerive P, Monté D, Dubois G, Trottein F, Fruchart-Najib J,
29. Testini C, Smith RO, Jin Y, Martinsson P, Sun Y, Hedlund M, Mariani J, et al. The orphan nuclear receptor ROR alpha is a
et al. Myc-dependent endothelial proliferation is controlled by negative regulator of the inflammatory response. EMBO Rep. (2001)
phosphotyrosine 1212 in VEGF receptor-2. EMBO Rep. (2019) 2:42–8. doi: 10.1093/embo-reports/kve007
20:e47845. doi: 10.15252/embr.201947845 47. Liu Y, Chen Y, Zhang J, Liu Y, Zhang Y, Su Z. Retinoic acid receptor-
30. Jambusaria A, Hong Z, Zhang L, Srivastava S, Jana A, Toth PT, related orphan receptor α stimulates adipose tissue inflammation by
et al. Endothelial heterogeneity across distinct vascular beds during modulating endoplasmic reticulum stress. J Biol Chem. (2017) 292:13959–
homeostasis and inflammation. Elife. (2020) 9:e51413. doi: 10.7554/eLife. 69. doi: 10.1074/jbc.M117.782391
51413 48. Taylor AM, Humphries JM, White RM, Murphey RD, Burns CE, Zon
31. Saygin D, Tabib T, Bittar HET, Valenzi E, Sembrat J, Chan SY, et al. LI. Hematopoietic defects in rps29 mutant zebrafish depend upon p53
Transcriptional profiling of lung cell populations in idiopathic pulmonary activation. Exp Hematol. (2012) 40:228–37.e5. doi: 10.1016/j.exphem.2011.
arterial hypertension. Pulm Circ. (2020) 10:1–15. doi: 10.1177/20458940209 11.007
08782 49. Mirabello L, Macari ER, Jessop L, Ellis SR, Myers T, Giri N, et al. Whole-
32. Ghosh S, MarElia-Bennett CB, Hildreth BE, Lefler JE, Sharma SM, exome sequencing and functional studies identify RPS29 as a novel gene
Ostrowski MC. Redundant function of Ets1 and Ets2 in regulating M- mutated in multicase Diamond-Blackfan anemia families. Blood. (2014)
phase progression in post-natal angiogenesis. bioRxiv [Preprint]. (2020). 124:24–32. doi: 10.1182/blood-2013-11-540278
doi: 10.1101/2020.02.19.956417 50. Zhuo L-A, Wen Y-T, Wang Y, Liang Z-F, Wu G, Nong M-D, et al. LncRNA
33. Harris TA, Yamakuchi M, Kondo M, Oettgen P, Lowenstein CJ. Ets-1 and Ets- SNHG8 is identified as a key regulator of acute myocardial infarction by RNA-
2 regulate the expression of MicroRNA-126 in endothelial cells. Arterioscler seq analysis. Lipids Health Dis. (2019) 18:201. doi: 10.1186/s12944-019-1142-0
Thromb Vasc Biol. (2010) 30:1990–7. doi: 10.1161/ATVBAHA.110. 51. Gao L, Gao J, Liang Y, Li R, Xiao Q, Zhang Z, et al. Integration
211706 analysis of a miRNA-mRNA expression in A549 cells infected with a
34. Gutierrez KD, Morris VA, Wu D, Barcy S, Lagunoff M. Ets-1 is novel H3N2 swine influenza virus and the 2009. H1N1 pandemic influenza
required for the activation of VEGFR3 during latent kaposi’s sarcoma- virus. Infect Genet Evol. (2019) 74:103922. doi: 10.1016/j.meegid.2019.
associated herpesvirus infection of endothelial cells. J Virol. (2013) 87:6758– 103922
68. doi: 10.1128/JVI.03241-12 52. Wright H, Matthews J, Chapple I, Ling-Mountford N, Cooper P.
35. Herkt CE, Caffrey BE, Surmann K, Blankenburg S, Gesell Salazar M, Jung Periodontitis associates with a type 1 IFN signature in peripheral blood
AL, et al. A MicroRNA network controls legionella pneumophila replication neutrophils. J Immunol. (2008) 181:5775–84. doi: 10.4049/jimmunol.181.
in human macrophages via LGALS8 and MX1. mBio. (2020) 11:e03155- 8.5775
19. doi: 10.1128/mBio.03155-19 53. Pan J, Yu H, Guo Z, Liu Q, Ding M, Xu K, et al. Emerging role of
36. Cattaneo V, Tribulatti MV, Carabelli J, Carestia A, Schattner M, sperm-associated antigen 9 in tumorigenesis. Biomed Pharmacother. (2018)
Campetella O. Galectin-8 elicits pro-inflammatory activities in the 103:1212–6. doi: 10.1016/j.biopha.2018.04.168
endothelium. Glycobiology. (2014) 24:966–73. doi: 10.1093/glycob/ 54. Montagnac G, Sibarita J-B, Loubéry S, Daviet L, Romao M, Raposo G,
cwu060 et al. ARF6 interacts with JIP4 to Control a motor switch mechanism
37. Sarmento L, Afonso CL, Estevez C, Wasilenko J, Pantin-Jackwood regulating endosome traffic in Cytokinesis. Curr Biol. (2009) 19:184–
M. Differential host gene expression in cells infected with 95. doi: 10.1016/j.cub.2008.12.043
highly pathogenic H5N1 avian influenza viruses. Vet Immunol 55. Ren B, Wei X, Zou G, He J, Xu G, Xu F, et al. Cancer testis antigen SPAG9 is a
Immunopathol. (2008) 125:291–302. doi: 10.1016/j.vetimm.2008. promising marker for the diagnosis and treatment of lung cancer. Oncol Rep.
05.021 (2016) 35:2599–605. doi: 10.3892/or.2016.4645
38. Sharma A, Markey M, Torres-Muñoz K, Varia S, Kadakia M, Bubulya A, et al. 56. Dixon DM, Choi J, El-Ghazali A, Park SY, Roos KP, Jordan MC, et al. Loss of
Son maintains accurate splicing for a subset of human pre-mRNAs. J Cell Sci. muscleblind-like 1 results in cardiac pathology and persistence of embryonic
(2011) 124:4286–98. doi: 10.1242/jcs.092239 splice isoforms. Sci Rep. (2015) 5:9042. doi: 10.1038/srep09042
39. Lu X, Ng HH, Bubulya PA. The role of SON in splicing, development, 57. Jones DP, True HD, Patel J. Leukocyte trafficking in cardiovascular
and disease. Wiley Interdiscip Rev RNA. (2014) 5:637–46. doi: 10.1002/ disease: insights from experimental models. Mediators Inflamm. (2017)
wrna.1235 2017:9746169. doi: 10.1155/2017/9746169
40. Schroeder AM, Allahyari M, Vogler G, Missinato MA, Nielsen T, Yu 58. Anderson R, Lagnado A, Maggiorani D, Walaszczyk A, Dookun
MS, et al. Model system identification of novel congenital heart disease E, Chapman J, et al. Length-independent telomere damage
gene candidates: focus on RPL13. Hum Mol Genet. (2019) 28:3954– drives post-mitotic cardiomyocyte senescence. EMBO J. (2019)
69. doi: 10.1093/hmg/ddz213 38:e100492. doi: 10.15252/embj.2018100492
41. Pravata VM, Muha V, Gundogdu M, Ferenbach AT, Kakade PS, Vandadi 59. Wang H, Sun X, Chou J, Lin M, Ferrario CM, Zapata-Sudo G, et al.
V, et al. Catalytic deficiency of O-GlcNAc transferase leads to X- Inflammatory and mitochondrial gene expression data in GPER-deficient
linked intellectual disability. Proc Natl Acad Sci USA. (2019) 116:14961– cardiomyocytes from male and female mice. Data Brief. (2016) 10:465–
70. doi: 10.1073/pnas.1900065116 73. doi: 10.1016/j.dib.2016.11.057

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60. Ranek MJ, Stachowski MJ, Kirk JA, Willis MS. The role of heat shock proteins Copyright © 2020 Hachim, Al Heialy, Senok, Hamid and Alsheikh-Ali. This is an
and co-chaperones in heart failure. Philos Trans R Soc Lond B Biol Sci. (2018) open-access article distributed under the terms of the Creative Commons Attribution
373:1738. doi: 10.1098/rstb.2016.0530 License (CC BY). The use, distribution or reproduction in other forums is permitted,
provided the original author(s) and the copyright owner(s) are credited and that the
Conflict of Interest: The authors declare that the research was conducted in the original publication in this journal is cited, in accordance with accepted academic
absence of any commercial or financial relationships that could be construed as a practice. No use, distribution or reproduction is permitted which does not comply
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