Mechanism of Rotavirus Entry

NIH Pandemic-Era Grants

Pandemic Era Grants

2024

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Principal Investigator: Siyuan  Ding
Organization: WASHINGTON UNIVERSITY
Fiscal Year: 2024
Award: $194,375
Funding agency: National Institute of Allergy and Infectious Diseases

Rotavirus (RV) is a major etiological cause of severe gastroenteritis and diarrhea worldwide, resulting in the death of more than 200,000 children each year. Current RV vaccines have limited efficacy in endemic countries and no direct antivirals are available. Our overall objective is to better understand the mechanism by which human RVs enter human small bowel intestinal epithelial cells (IECs). The head region (VP8*) of the RV outer capsid spike protein VP4 is responsible for viral entry. However, the identities of host factors that interact with VP8* and their roles in human RV entry is unclear. In preliminary studies, we conducted a flow cytometry-based genome-scale CRISPR/Cas9 knockout screen and identified several transmembrane host proteins as putative attachment factors. In Aim 1, we will use CRISPR/Cas9 gene editing and classic adsorption assays to functionally examine the role of these candidates in human RV entry into HT-29 cells and primary human IECs. In Aim 2, we will employ chemical labeling and proteomic approaches and identify the VP8*-interacting cellular partners during human RV entry. Using host genetic screens, RV reverse genetics, and human intestinal organoid cultures, we expect that our study will elucidate the molecular mechanisms by which host molecules facilitate the early replication of human RVs. We ultimately aim to harness that knowledge to develop countermeasures to impede infection and protect against diarrheal diseases.

Terms: <0-11 years old><Adsorption><Affinity Chromatography><Age><Amino Acids><Animal Model><Animal Models and Related Studies><Anti-viral Agents><Antigenic Determinants><Antigens><Apical><Assay><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Binding><Binding Determinants><Binding Sites><Bioassay><Biological Assay><Biotin><Blood Group Antigens><CRISPR approach><CRISPR based approach><CRISPR editing screen><CRISPR method><CRISPR methodology><CRISPR screen><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based screen><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 screen><CRISPR/Cas9 technology><Capsid><Cas nuclease technology><Causality><Cell Body><Cell Isolation><Cell Line><Cell Segregation><Cell Separation><Cell Separation Technology><CellLine><Cells><Cessation of life><Chemicals><Child><Child Youth><Children (0-21)><Clinical><Clinical Treatment Moab><Closure by Ligation><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Co-Immunoprecipitations><Colon><Combining Site><Complex><Country><Crystallographies><Crystallography><Cy5><Data><Death><Dependence><Diarrhea><Distant><EXTL3><EXTL3 gene><EXTR1><Enterocytes><Enteroendocrine Cell><Epidemiology><Epithelial Cells><Epitopes><Escape Mutant><Etiology><Event><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Gastroenteritis><Genes><Genetic Screening><Glycans><HT-29><HT-29 Cells><HT29 Cells><Head><Heparan Sulfate><Heparitin Sulfate><High Throughput Assay><Host Factor><Host Factor Protein><Hu-mABs><Human><Incubated><Infant><Infection><Integral Membrane Protein><Integration Host Factors><Intestinal><Intestines><Intrinsic Membrane Protein><KIAA0519><Knock-out><Knockout><Knowledge><Label><Ligation><Lymphatic cell><Lymphocyte><Lymphocytic><Maps><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Membrane><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Modern Man><Molecular><Molecular Interaction><Monkeys><Monoclonal Antibodies><Morbidity><Morbidity - disease rate><N-Acetylneuraminic Acids><Norovirus><Norwalk-like Viruses><Organoids><Outcome><Persons><Polysaccharides><Preventative vaccine><Preventive vaccine><Prophylactic vaccine><Proteins><Proteomics><Reactive Site><Receptor Protein><Recombinants><Reporter><Role><Rotavirus><Rotavirus Infections><Rotavirus Vaccines><Route><Sialic Acids><Side><Small Intestines><Strains Cell Lines><Structural Protein><Surface><Surface Proteins><System><Therapeutic><Transmembrane Protein><Transmembrane Protein Gene><Transmission Electron Microscopy><Tropism><Vaccines><Viral><Virion><Virus Particle><Vitamin H><affinity purification><ages><aminoacid><anti-viral compound><anti-viral drugs><anti-viral medication><anti-viral therapeutic><anti-virals><bowel><causation><cell sorting><clustered regularly interspaced short palindromic repeats screen><coenzyme R><cultured cell line><cyanine dye 5><data modeling><design><designing><diarrheal disease><diarrheal illness><disease causation><epidemiologic><epidemiological><flow cytophotometry><genome scale><genome-wide><genomewide><high throughput screening><humAbs><human mAbs><human monoclonal antibodies><human monoclonals><immunogen><improved><in vivo><in vivo Model><infancy><infantile><innovate><innovation><innovative><intestinal epithelium><kidney epithelial cell><kids><lymph cell><mAbs><membrane structure><model of animal><model of data><model the data><modeling of the data><monoclonal Abs><monolayer><mortality><mouse model><murine model><neutralizing mAb><neutralizing monoclonal antibodies><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><new vaccines><next generation therapeutics><next generation vaccines><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><novel vaccines><rational design><receptor><reverse genetics><small bowel><social role><suckling><tool><youngster>