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Principal Investigator: Santimukul Santra
Organization: PITTSBURG STATE UNIVERSITY
Fiscal Year: 2022
Award: $76,946
Funding agency: National Institute of General Medical Sciences
Program Director/Principal Investigator: Santra, Santimukul
PROJECT SUMMARY/ABSTRACT
This application proposes to develop liposome-coated iron oxide nanosensors (LIONs) mimicking as
magnetically-labeled host membranes to detect fusion interactions of enveloped viruses. Spin-spin T2 relaxation
technique will be established as a generic bioanalytical method to detect dynamic conformational changes in
viral glycoproteins that play vital roles in fusion interactions. Within the first aim, influenza fusion protein
interactions with LIONs membrane will be studied, considering diverse fusion triggering environmental
factors including low pH, protease activation and receptor binding. Further studies to investigate the
sensitivity of this LIONs technology in quantifying similar fusion interactions with more native configurations of
HA is demonstrated using virus-like particle (VLPs) and shorter domains derived from HA. Similarly, fusion
experiments will be carried out using ganglioside-conjugated LIONs. The promising outcome of this research will
be screening potential antiviral candidates including small molecules and neutralizing antibodies.
The second aim establishes the broad adaptability of this sensitive LIONs technology for evaluation of fusion
interactions of enveloped viruses where presence of membrane receptor is an important trigger, for example,
SARS-CoV-2. Receptor-conjugated LIONs (R-LIONs) will be developed for the elucidation of this vital process
using spike proteins and reporter virus particles (RVPs) of SARS-CoV-2. R-LIONs offers a novel approach of
exploring the role of variety of entry receptors that may play an important role in SARS-CoV-2 membrane fusion.
The third aim of this proposal determines further advantage of nanosensor technology for the real-time
monitoring of virus-mediated cell fusion. SARS-CoV-2 virus entry into ACE2 receptor overexpressing mammalian
cells (HEK-293T) will be examined by developing magnetically labeled reporter virus particles (M-RVPs). Parallel
experiments will be carried out in the presence of fusion inhibitors and neutralizing antibodies. The important
application of this experiment will be demonstrated by screening potential new fusion inhibitors and antiviral
candidates.
The proposed AREA proposal, if successful, will develop novel nanosensor technology for the rapid detection of
fusion interactions of enveloped viruses including SARS-CoV-2 and influenza. This adaptable technology will
allow for rapid screening of potential drug candidates and fusion inhibitors of many other enveloped viruses, in
a timely fashion. Most importantly, this AREA proposal will initiate the proposed PURE program for
undergraduate education, and increase the hands-on biomedical research opportunity and learning for students
in our undergraduate-focused institution.
OMB No. 0925-0001 and 0925-0002 (Rev. 03/20 Approved Through 02/28/2023) Page Project Summary
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><ACE2><Affinity><Antiviral Agents><Antiviral Drugs><Antivirals><Assay><Bioassay><Biologic Assays><Biological><Biological Assay><Biomedical Research><COVID-19 virus><COVID19 virus><Cell Body><Cell Communication and Signaling><Cell Line><Cell Membrane Lipids><Cell Signaling><Cell fusion><Cell membrane><CellLine><Cells><Characteristics><Chimera Protein><Chimeric Proteins><CoV-2><CoV2><Coloring Agents><Coupling><Cytoplasmic Membrane><Data><Data Analyses><Data Analysis><Development><Disease Outbreaks><Dyes><Ebola><Environmental Factor><Environmental Risk Factor><Esteroproteases><Evaluation><Fe oxide><Fluorescence><Fusion Protein><G(A(2)) Ganglioside><G(M1) Ganglioside><GA(2) Ganglioside><Ganglioside GM1><Gangliosides><Glycoproteins><Goals><Grippe><Hemagglutinin><Image Analyses><Image Analysis><Infection><Influenza><Influenza Virus><Institution><Intracellular Communication and Signaling><Investigation><Label><Learning><Lipids><Liposomal><Liposomes><Luciferase Immunologic><Luciferases><Magnetism><Mammalian Cell><Measurement><Measures><Mediating><Membrane><Membrane Fusion><Membrane Lipids><Methods><Modeling><Molecular Configuration><Molecular Conformation><Molecular Stereochemistry><Monosialosyl Tetraglycosyl Ceramide><N-Acetylneuraminic Acids><Outbreaks><Outcomes Research><Pathogen detection><Pathogenicity><Peptidases><Peptide Hydrolases><Phase><Plasma Membrane><Play><Principal Investigator><Process><Property><Protease Gene><Proteases><Proteinases><Proteins><Proteolytic Enzymes><Rapid screening><Reaction><Receptor Protein><Recombinants><Relaxation><Relaxation Technics><Relaxation Techniques><Reporter><Reporting><Reproducibility><Role><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><Screening procedure><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><Sialic Acids><Sialoglycosphingolipids><Signal Transduction><Signal Transduction Systems><Signaling><Strains Cell Lines><Students><Technology><Temperature><Testing><Time><Tripcellim><Trypsin><Validation><Viral><Viral Fusion Proteins><Viral Gene Products><Viral Gene Proteins><Viral Pathogenesis><Viral Proteins><Virion><Virus><Virus Particle><Virus-like particle><Work><Wuhan coronavirus><ZIKA><analytical method><angiotensin converting enzyme 2><angiotensin converting enzyme II><anti-viral agents><anti-viral compound><anti-viral drugs><anti-viral medication><anti-viral therapeutic><anti-virals><antiviral compound><antiviral medication><antiviral therapeutic><base><biologic><biological signal transduction><colcothar><conformation><conformational state><coronavirus disease 2019 virus><coronavirus disease-19 virus><cultured cell line><data interpretation><developmental><drug candidate><environmental risk><experiment><experimental research><experimental study><ferric oxide><hCoV19><high throughput technology><image evaluation><image interpretation><influenzavirus><inhibitor><insight><instrument><iron oxide><magnetic><membrane structure><metal chelator><nCoV2><nano sensing><nano sensors><nanosensing><nanosensors><neutralizing antibody><new approaches><new technology><novel><novel approaches><novel strategies><novel strategy><novel technologies><overexpress><overexpression><pandemic><pandemic disease><plasmalemma><programs><rapid detection><real time monitoring><realtime monitoring><receptor><receptor binding><receptor bound><red iron oxide><screening><screening tools><single molecule><small molecule><social role><temporal measurement><temporal resolution><time measurement><time use><tool><undergrad><undergraduate><undergraduate education><undergraduate student><virus envelope><virus pathogenesis><virus protein><virus-like nanoparticles><viruslike particle>