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Principal Investigator: PAUL L FOX
Organization: CLEVELAND CLINIC LERNER COM-CWRU
Fiscal Year: 2021
Award: $402,500
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
Project Summary/Abstract
Obesity is an epidemic-scale problem in the U.S. affecting about 35% of the adult population, and is a major risk
factor for the ongoing pandemic, COVID-19. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is
the RNA betacoronavirus that is the causative agent of COVID-19. Despite rapid progress in developing
effective vaccines, progress in devising improved therapeutics has been slow, and there is an urgent need for
anti-viral therapeutics for treatment of infected patients not protected by vaccines. Certain demographics are
variably resistant to vaccination, for example, obesity markedly reduces effectiveness of several vaccines.
Therapeutics can also be critical in the eventuality that mutations in the virus render the vaccine ineffective. The
molecular events responsible for expression of SARS-CoV-2 proteins are known from studies of other
betacoronaviruses; however, the regulatory pathways and pathological conditions determining their expression
are poorly understood. The translation of viral RNA utilizes the host mRNA translation machinery, primarily
regulated by specific RNA-binding proteins or complexes that bind sequence or structural elements in terminal
non-coding regions. Importantly, the coding regions of SARS-CoV-2 genomic RNA and the ten subgenomic
mRNAs (sgmRNAs) are likewise bordered by non-coding upstream and downstream regions, termed the 5'-
leader and 3'-end sequences, respectively. A critical feature of the genome and the sgmRNAs of SARS-CoV-2
is that identical 5'-leader and 3'-end sequences are present in all. Thus, the terminal sequences represent
novel, unexplored targets for interference with virus assembly and function. We have discovered a 39-nt
sequence in the 3'-end of SARS-CoV-2 bearing structural similarity to the GAIT (interferon-gamma-activated
inhibitor of translation) RNA element previously described by us. We show that glutamyl-prolyl tRNA synthetase
(EPRS) a protein that binds the human GAIT element also binds the vGLE. Moreover, IFN-γ, a pro-inflammatory
cytokine, and insulin, an obesity-induced hormone, markedly increases expression of a luciferase reporter
bearing the intact vGLE. These results are the first to show a functional consequence of an RNA element in the
3'-end sequence of SARS-CoV-2. We hypothesize that binding of EPRS to the vGLE stimulates sgmRNA
translation required for expression of structural and other SARS-CoV-2 proteins, and for programmed ribosomal
frameshifting required for genome replication. We will test this hypothesis by pursuing two Specific Aims: In Aim
1 we elucidate the role of EPRS binding to the SARS-CoV-2 3'UTR vGLE in regulating viral replication and
sgmRNA translation. In Aim 2 we develop RNA inhibitors that target EPRS/vGLE interactions and block viral
protein expression. We anticipate these fundamental studies will provide the first information on the function of
the 3'-end of SARS-CoV-2, and will provide a foundation for development of therapeutic agents to be used in
combination with mechanistically distinct anti-viral agents targeting the vGLE and possibly emerging
betacoronaviruses.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><21+ years old><3' Untranslated Regions><3'UTR><ACE2><Acute><Adipocytes><Adipose Cell><Adipose tissue><Adult><Adult Human><Affect><Affinity><Allergy><Anti-Sense RNA><Antigenic Determinants><Antisense RNA><Antiviral Agents><Antiviral Drugs><Antivirals><Assay><Binding><Binding Determinants><Binding Proteins><Bioassay><Biologic Assays><Biological Assay><COVID><COVID crisis><COVID epidemic><COVID pandemic><COVID-19><COVID-19 crisis><COVID-19 epidemic><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 pandemic><COVID-19 public health crisis><COVID-19 therapeutics><COVID-19 virus><COVID19><COVID19 crisis><COVID19 epidemic><COVID19 global health crisis><COVID19 global pandemic><COVID19 health crisis><COVID19 pandemic><COVID19 public health crisis><COVID19 therapeutics><COVID19 virus><CV-19><CV19><Cell Surface Receptors><Cleaved cell><CoV disease><CoV-2><CoV2><Code><Coding System><Complex><Coronaviridae><Coronavirus><EC 2.7.7.48><EPRS enzyme><Effectiveness><Elements><Endocrine Gland Secretion><Epidemic><Epitopes><Event><Fat Cells><Fatty Tissue><Foundations><Functional RNA><G(s), alpha Subunit><G(s), α Subunit><G(s)alpha><G(s)α><GTP-Binding Protein alpha Subunits, Gs><GTP-Binding Protein α Subunits, Gs><Gait><Gamma interferon><Genetic Alteration><Genetic Change><Genetic Translation><Genetic defect><Genome><Genomics><Gs alpha Family G-Protein><Gsα><Gαs><Hepatitis B><Hepatitis B Infection><Hormones><Human><Humulin R><Hypersensitivity><IFN-Gamma><IFN-g><IFN-γ><IFNG><IFNγ><Immune Interferon><Incidence><Inflammatory><Influenza B><Influenza B Virus><Influenza Viruses Type B><Insulin><Interferon Gamma><Interferon Type II><Interferon-gamma><Knowledge><Ligand Binding Protein><Ligand Binding Protein Gene><Lipocytes><Luciferase Immunologic><Luciferases><Mature Lipocyte><Mature fat cell><Messenger RNA><Modern Man><Molecular><Molecular Interaction><Mutate><Mutation><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Novolin R><Obesity><Oligo><Oligonucleotides><Orthomyxoviruses Type B><Pathologic><Pathway interactions><Patients><Polyproteins><Population><Protein Binding><Proteins><Proteomics><RNA><RNA Gene Products><RNA Replicase><RNA-Binding Proteins><RNA-Dependent RNA Polymerase><RNA-Directed RNA Polymerase><Receptor Protein><Regular Insulin><Regulation><Regulatory Ns Protein><Regulatory Pathway><Reporter><Resistance><Ribonucleic Acid><Ribosomal Frame Shifting><Ribosomal Frameshift><Ribosomal Frameshifting><Risk Factors><Role><SARS Virus><SARS corona virus><SARS corona virus 2><SARS coronavirus><SARS-Associated Coronavirus><SARS-CoV><SARS-CoV-2><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 inhibitor><SARS-CoV-2 pandemic><SARS-CoV-2 therapeutics><SARS-CoV2><SARS-CoV2 epidemic><SARS-CoV2 pandemic><SARS-Related Coronavirus><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><SARS-coronavirus-2 therapeutics><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Series><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><Severe Acute Respiratory Syndrome Virus><Severe Acute Respiratory Syndrome corona virus><Severe Acute Respiratory Syndrome coronavirus><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 inhibitor><Severe acute respiratory syndrome coronavirus 2 pandemic><Severe acute respiratory syndrome coronavirus 2 therapeutics><Severe acute respiratory syndrome related corona virus 2><Stimulatory Gs G-Protein><Stimulus><Structural Protein><Structure><Syndrome><Testing><Therapeutic><Therapeutic Agents><Therapeutic Hormone><Therapeutic Intervention><Translational Frameshifting><Translations><UTRs><Untranslated RNA><Untranslated Regions><Vaccination><Vaccines><Viral><Viral Gene Products><Viral Gene Proteins><Viral Genome><Viral Hepatitis B><Viral Proteins><Virus><Virus Assembly><Virus Replication><Wuhan coronavirus><adipose><adiposity><adulthood><alpha Subunit Stimulatory GTP-Binding Protein><alpha-Gs><angiotensin converting enzyme 2><angiotensin converting enzyme II><anti-viral agents><anti-viral drugs><anti-virals><base><beta CoV><beta coronavirus><betaCoV><betacoronavirus><block SARS-CoV-2><block severe acute respiratory syndrome coronavirus 2><bound protein><cleaved><corona virus><corona virus disease><corona virus disease 2019><corona virus disease 2019 epidemic><corona virus disease 2019 pandemic><coronavirus disease><coronavirus disease 2019><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global pandemic><coronavirus disease 2019 health crisis><coronavirus disease 2019 pandemic><coronavirus disease 2019 public health crisis><coronavirus disease 2019 therapeutics><coronavirus disease 2019 virus><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease pandemic><corpulence><cytokine><demographics><experiment><experimental research><experimental study><genome mutation><genomic RNA><glutamyl-prolyl-tRNA synthetase><hCoV19><improved><infection with HBV><infection with hepatitis B virus><inhibit SARS-CoV-2><inhibit severe acute respiratory syndrome coronavirus 2><inhibitor><inhibitor/antagonist><intervention therapy><lFN-Gamma><mRNA><mRNA Translation><mimetics><nCoV2><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><noncoding><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><obese patients><oligos><pandemic><pandemic disease><particle><pathway><patients with obesity><protein expression><receptor><resistant><serum hepatitis><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><severe acute respiratory syndrome-CoV><social role><targeted agent><therapeutic agent development><therapeutic development><therapeutic target><therapeutics against COVID-19><therapeutics against COVID19><therapeutics against SARS-CoV-2><therapeutics against SARS-coronavirus-2><therapeutics against Severe acute respiratory syndrome coronavirus 2><therapeutics against coronavirus disease 2019><therapeutics for novel coronavirus><viral RNA><viral assembly><viral multiplication><viral replication><virus RNA><virus genome><virus multiplication><virus protein><white adipose tissue><yellow adipose tissue><α-Gs><β CoV><β coronavirus><βCoV>