The world is immersed in a health crisis rivaled only by the Spanish flu pandemic of 1918-1919. The difference between the crisis of today and the crisis a century ago is that we have advanced communication technology greatly so that huge populations of people around the globe are aware of risks and can take appropriate precautions, such as the employment of quarantining, isolation, social distancing, and masks and handwashing. Governments have closed schools and outlawed large social gatherings. Medical care has greatly improved, and those most affected have received symptomatic treatment in the absence of a cure for CoVID-19. The problem must be dealt at multiple fronts, such as vaccine development, drug development and new technologies, assays for mitigating viral effects. The proposed project is significant because it proposes a novel in vitro biophysical screening assay for a unique and yet untapped RNA structure in SARS-CoV-2 virus, that can be used in the future to generate RNA specific antiviral compounds. Nucleic acids are promising avenues for drug design, both as therapeutics and as targets. Here we propose an innovative screening approach for identification of a novel class of ligands that are specific for an RNA element within the viral RNA genome that is vital for replication of the virus, and we propose a biophysical screening assay as a first step for identifying such ligands. First, as outlined in Specific Aim 1, we will characterize a model nucleic acid (RNA) domain that will be synthesized, characterized and used to identify a RNA specific fluorescent probe. The optimized probe will then be used for developing a high-throughput screening assay for discovery of nanomolar binders to this RNA. The RNA specific high-affinity binders will then be combined with sequence-specific RNA binding ligands to validate the assay development and its utility. The mechanism of action will be confirmed using inhibition of firefly luciferase translation in a reporter gene assay (Specific Aim 2). While the focus of this application, as the needed first step, is on the development and validation of the biophysical assay for the underlying SARS-CoV-2 RNA structures, a successful validation of the approach will open the doors for discovery and development of lead compounds for inhibition of SARS-CoV in Phase II studies. NUBAD and its team of scientists and collaborators is uniquely qualified to perform these assays and develop the potential leads in Phase II, in addition to providing a template for the scientific community to use the screening technology for their own discovery platforms. Success of the approach will also allow us to expand the screening technology to other RNA structures in SARS and other RNA viruses, and provide the screening resource as a service to the scientific community.
Public Health Relevance Statement: PROJECT NARRATIVE The global CoVID-19 pandemic of 2019-2020 has had immeasurable toll on human life across the globe, affecting individuals on a personal social, physical, and mental health level, and likewise impacting national and global economies and overloading an already burdened national health care system. In order to combat this world-wide problem, new targets and innovative strategies for antiviral drug design must be implemented. This project addresses this emergent problem by targeting a regulatory RNA sequence in SARS-CoV-2 with the help of a novel invitro biophysical screening assay to identify compounds that can potentially block viral RNA replication.
Project Terms: Affect; Amino Sugars; aminosugar; Antiviral Agents; Antiviral Drugs; Antivirals; anti-viral agents; anti-viral compound; anti-viral drugs; anti-viral medication; anti-viral therapeutic; anti-virals; antiviral compound; antiviral medication; antiviral therapeutic; Awareness; Biological Assay; Assay; Bioassay; Biologic Assays; Biophysics; biophysical foundation; biophysical principles; biophysical sciences; Cell Culture Techniques; cell culture; cell cultures; Chemistry; High Pressure Liquid Chromatography; HPLC; High Performance Liquid Chromatography; High Speed Liquid Chromatography; Communication; Communities; Coughing; Cessation of life; Death; Disease; Disorder; Drug Design; Elements; Employment; Fluorescence; Fluorescent Probes; Future; Genes; Bacterial Genes; Genome; Government; Grant; Handwashing; hand washing; Health; Healthcare Systems; Health Care Systems; Health Status; Level of Health; Human; Modern Man; In Vitro; Infection; Laboratories; Lead; Pb element; heavy metal Pb; heavy metal lead; Libraries; Ligands; Masks; Mental Health; Mental Hygiene; Psychological Health; Persons; Nucleic Acids; Patents; Legal patent; Proteins; Public Health; isolation/quarantine; Quarantine; Research Resources; Resources; Reverse Transcription; Risk; Non-Polyadenylated RNA; RNA Gene Products; Ribonucleic Acid; RNA; RNA Viruses; mRNA; Messenger RNA; virus RNA; viral RNA; social role; Role; Schools; Social Distance; Technology; Tennessee; Transactivation; trans-activation; Genetic Transcription; Gene Transcription; RNA Expression; Transcription; Translations; Universities; Vaccination; Vaccines; Viral Proteins; Viral Gene Products; Viral Gene Proteins; virus protein; Virus Replication; viral multiplication; viral replication; virus multiplication; Virus; Work; Antisense Oligonucleotides; Anti-Sense Oligonucleotides; Antisense Agent; anti-sense agent; anti-sense oligo; antisense oligo; Caring; RNA Sequences; improved; Solid; Phase; Medical; Reporter Genes; Chemicals; Individual; analog; Therapeutic; Photinus luciferin 4 monooxygenase; Firefly Luciferases; Exposure to; Life; Scientist; Complex; Event; Country; Respiratory distress; physical health; physical conditioning; Viral; respiratory; interest; Prevent infection; Infection prevention; Services; mutant; success; develop a vaccine; develop vaccines; development of a vaccine; vaccine development; RNA library; tech development; technology development; microbial; 5'UTR; mRNA Leader Sequences; 5' Untranslated Regions; Structure; novel; novel technologies; new technology; social; Congestive; Proteome; Modeling; assay development; drug development; High Throughput Assay; high throughput screening; RNA Regulatory Sequences; Regulatory RNA Region; Regulatory Ribonucleic Acid Region; Ribonucleic Acid Regulatory Sequences; RNA bound; RNA Binding; Molecular Interaction; Binding; PMO oligomer; phosphorodiamidate morpholino oligomer; SARS; SARS coronavirus disease; SARS-CoV disease; Severe Acute Respiratory Syndrome CoV disease; Severe Acute Respiratory Syndrome coronavirus disease; Severe Acute Respiratory Syndrome; SARS Virus; SARS corona virus; SARS-Associated Coronavirus; SARS-CoV; SARS-CoV-1; SARS-Related Coronavirus; Severe Acute Respiratory Coronavirus; Severe Acute Respiratory Syndrome Virus; Severe Acute Respiratory Syndrome corona virus; Severe Acute Respiratory Syndrome coronavirus; severe acute respiratory syndrome-CoV; SARS coronavirus; preventing; prevent; RNA replication; Address; Affinity; Interruption; NIAID; National Institute of Allergy and Infectious Disease; Ligand Binding; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Small Business Technology Transfer Research; STTR; Transcript; Validation; Development; developmental; symptom management; manage symptom; pandemic disease; pandemic; pandemic influenza; flu virus pandemic; influenza virus pandemic; pandemic flu; pandemic strain of influenza; design; designing; Population; innovation; innovate; innovative; stem; combat; phase 2 study; phase II study; screening; genomic RNA; biophysical analysis; biophysical studies; nanomolar; nano-molar; symptom treatment; symptomatic treatment; treat symptom; experimental study; experiment; experimental research; Spanish flu; Spanish influenza; COVID-19; COVID19; CV-19; CV19; corona virus disease 2019; coronavirus disease 2019; coronavirus disease-19; coronavirus infectious disease-19; 2019-nCoV; 2019 novel corona virus; 2019 novel coronavirus; COVID-19 virus; COVID19 virus; CoV-2; CoV2; SARS corona virus 2; SARS-CO-V2; SARS-COVID-2; SARS-CoV-2; SARS-CoV2; SARS-associated corona virus 2; SARS-associated coronavirus 2; SARS-coronavirus-2; SARS-related corona virus 2; SARS-related coronavirus 2; SARSCoV2; Severe Acute Respiratory Coronavirus 2; 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-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 related corona virus 2; Wuhan coronavirus; coronavirus disease 2019 virus; coronavirus disease-19 virus; hCoV19; nCoV2; COVID-19 pandemic; COVID crisis; COVID epidemic; COVID pandemic; COVID-19 crisis; COVID-19 epidemic; COVID-19 global health crisis; COVID-19 global pandemic; COVID-19 health crisis; COVID-19 public health crisis; COVID19 crisis; COVID19 epidemic; COVID19 global health crisis; COVID19 global pandemic; COVID19 health crisis; COVID19 pandemic; COVID19 public health crisis; SARS-CoV-2 epidemic; SARS-CoV-2 global health crisis; SARS-CoV-2 global pandemic; SARS-CoV-2 pandemic; SARS-CoV2 epidemic; SARS-CoV2 pandemic; SARS-coronavirus-2 epidemic; SARS-coronavirus-2 pandemic; Severe Acute Respiratory Syndrome CoV 2 epidemic; Severe Acute Respiratory Syndrome CoV 2 pandemic; Severe acute respiratory syndrome coronavirus 2 epidemic; Severe acute respiratory syndrome coronavirus 2 pandemic; corona virus disease 2019 epidemic; corona virus disease 2019 pandemic; 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 crisis; coronavirus disease epidemic; coronavirus disease pandemic; coronavirus disease-19 global pandemic; coronavirus disease-19 pandemic; severe acute respiratory syndrome coronavirus 2 global health crisis; severe acute respiratory syndrome coronavirus 2 global pandemic; coronavirus disease; COVID; CoV disease; corona virus disease; COVID-19 treatment; COVID-19 therapy; COVID19 therapy; COVID19 treatment; SARS-CoV-2 therapy; SARS-CoV-2 treatment; coronavirus disease 2019 therapy; coronavirus disease 2019 treatment; severe acute respiratory syndrome coronavirus 2 therapy; severe acute respiratory syndrome coronavirus 2 treatment; treat COVID-19; treat COVID19; treat SARS-CoV-2; treat coronavirus disease 2019; treat severe acute respiratory syndrome coronavirus 2; Rapid screening; SARS-CoV-2 infection; COVID-19 infection; COVID19 infection; SARS-CoV2 infection; Severe acute respiratory syndrome coronavirus 2 infection; coronavirus disease 2019 infection; infected with COVID-19; infected with COVID19; infected with SARS-CoV-2; infected with SARS-CoV2; infected with coronavirus disease 2019; infected with severe acute respiratory syndrome coronavirus 2; pandemic coronavirus; coronavirus of pandemic concern; coronavirus of pandemic potential; coronavirus with pandemic potential; pandemic threat coronavirus
