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Principal Investigator: Charles M Rice
Organization: ROCKEFELLER UNIVERSITY
Fiscal Year: 2024
Award: $846,776
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
The yellow fever 17D live-attenuated vaccine has been administered to millions of people and is a highly
successful and safe vaccine providing essentially life-long protection. However, the mechanisms of attenuation
are poorly understood, and rare individuals suffer from sometimes fatal severe adverse events (SAEs) including
yellow fever-like viscerotropic disease, or neurologic disease. Epidemics of this mosquito-borne disease continue
to occur in tropical and sub-tropical regions of South America and Africa, so understanding the mechanisms of
17D attenuation and the factors predisposing to SAEs is critical for development of safer vaccines and SAE
prevention or treatment strategies. Work of others and of ours in the prior funding period determined that type I
interferon (IFN) responses are greater after infection of cells with 17D compared to the virulent Asibi parental
strain. New preliminary data demonstrate that the prophylactic IFN-induced antiviral state is also more effective
at controlling 17D. Importantly, our work uncovered mutations in genes encoding type I IFN receptor chains
(IFNAR1 and IFNAR2) and autoantibodies against type I IFNs as factors causing SAEs after 17D vaccination,
thus linking the observed IFN phenotype with 17D attenuation in vivo. We also discovered a differential
requirement between 17D and Asibi, mediated by the viral NS4B protein, for the host factor TMEM41B, with
increased innate immune responses to 17D in TMEM41B KO cells. Prior studies have been hampered by the
disparate specific infectivities of 17D and Asibi, precluding meaningful comparisons in bulk cell-based assays.
New preliminary data demonstrates that 17D and Asibi harboring the 17D E protein (Asibi+17D E) have similar
specific infectivities, replicate with similar kinetics in the HepG2 hepatocyte cell line, and maintain previously
described differences in IFN transcription and secretion. Unexpectedly, levels of phosphorylated STAT1 are
similar in 17D- and Asibi+17D E-infected cells. In this project continuation, in Aim 1 17D and Asibi+17D E will be
used to examine pattern recognition receptor activation, including kinase activation and downstream
transcriptional responses using phosphoproteomics and RNASeq. The mechanism by which Asibi+17D E blocks
IFN secretion, the process by which STAT1 phosphorylation occurs and the consequences to infection and role
of individual ISGs in controlling 17D and Asibi infection will be elucidated. In Aim 2 interactions of Asibi and 17D
with both pro- and anti-viral host factors will be characterized through imaging-based methods allowing single
cell resolution. We will examine responses in the infected cell and the influence of individual genes (at genome
scale) on innate immunity and infection. Patient-specific mutations associated with SAEs will be functionally
interrogated as we have done previously. In Aim 3 the role of NS4B in differential TMEM41B requirements will
be elucidated using proteomic and mutational approaches. These studies will uncover critical host-pathogen
interactions that can be exploited to develop safer vaccines, SAE treatment or prevention approaches, and new
drug targets. The information gained can also be broadly applied to vaccine development for other flaviviruses.
Terms: <A549><Adverse reactions><Affinity Chromatography><Alpha-Beta-Omega Interferon Receptor-1><Amino Acid Substitution><Amino Acids><Antiviral Protein Alpha Type><Approaches to prevention><Assay><Attenuated><Attenuated Vaccines><Autoantibodies><Bioassay><Biological Assay><CRISPR><CRISPR/Cas system><Cell Body><Cell Cycle Kinetics><Cell Kinetics><Cell Line><Cell secretion><CellLine><Cells><Cellular Secretion><Clustered Regularly Interspaced Short Palindromic Repeats><Complex><Coupled><DNA Alteration><DNA Sequence Alteration><DNA mutation><Data><Development><Disease><Disorder><Disparate><Dose><E protein><Epidemic><Event><FISH Technic><FISH Technique><FISH analysis><FISH assay><Flavivirus><Fluorescence In Situ Hybridization><Fluorescent in Situ Hybridization><Funding><Gene Transcription><Genes><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Genetic mutation><Goals><Group B Arbovirus><Hep G2><HepG2><HepG2 cell line><Hepatic Cells><Hepatic Parenchymal Cell><Hepatocyte><Host Factor><Host Factor Protein><HuIFN-Alpha-Rec><IFN><IFNAR><IFNAR1><IFNAR1 gene><IFNAR2><IFNAR2 gene><IFNARB><IFNBR><IFRC><Immune><Immune response><Immunes><Immunocompetent><Immunological response><Individual><Infection><Infection prevention><Innate Immune Response><Innate Immunity><Integration Host Factors><Interferon Alpha-Beta Receptor Alpha Chain><Interferon Receptor><Interferon Type I><Interferons><Kinases><Kinetics><Knock-out><Knockout><Label><Libraries><Life><Link><Lipids><Live-attenuated Vaccine><Liver Cells><Lung><Lung Respiratory System><Lytotoxicity><Maps><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measures><Mediating><Messenger RNA><Mosquito-borne disease><Mosquito-borne infectious disease><Mutation><Native Immunity><Natural Immunity><Nervous System Diseases><Nervous System Disorder><Neurologic Disorders><Neurological Disorders><Non-Polyadenylated RNA><Non-Specific Immunity><Nonspecific Immunity><Orthoflavivirus><Outcome><Patients><Pattern recognition receptor><Persons><Phenotype><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Predisposing Factor><Prevent infection><Preventative strategy><Prevention><Prevention approach><Prevention strategy><Preventive strategy><Process><Protein Phosphorylation><Proteins><Proteomics><RNA><RNA Expression><RNA Gene Products><RNA Seq><RNA replication><RNA sequencing><RNAseq><Receptor Activation><Receptor Gene><Resolution><Ribonucleic Acid><Role><STAT1><STAT1 gene><STAT91><Safety><Sampling><Sequence Alteration><Serious Adverse Event><Severe Adverse Event><Signal Pathway><South Africa><South America><Strains Cell Lines><Testing><Transcript><Transcription><Translations><Transphosphorylases><Vaccination><Vaccines><Viral><Viral Diseases><Viral Gene Products><Viral Gene Proteins><Viral Proteins><Virulent><Virus><Virus Diseases><Virus Replication><Work><Yellow Fever><Yellow Fever Vaccine><Yellow Fever Virus Infection><Yellow fever virus><affinity purification><aminoacid><attenuate><attenuates><attenuation><autoimmune antibody><autoreactive antibody><clinical practice><cultured cell line><cytotoxicity><develop a vaccine><develop vaccines><development of a vaccine><developmental><fitness><genome mutation><genome scale><genome-wide><genomewide><genomic alteration><host response><ifnar1 gene product><image-based method><imaging method><imaging modality><immune competent><immune system response><immunoresponse><improved><in vivo><innate immune pathways><insight><live vaccine><live vaccines><mRNA><mimetics><mutant><mutation scanning><mutation screening><neurological disease><neurotropic><new approaches><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel approaches><novel drug target><novel druggable target><novel pharmacotherapy target><novel strategies><novel strategy><novel therapeutic target><novel therapy target><pathogen><phospho-proteomics><phosphoproteomics><prevent><preventing><prophylactic><pulmonary><reconstitute><reconstitution><resolutions><response><self reactive antibody><serious adverse experience><serious adverse reaction><social role><transcriptome sequencing><transcriptomic sequencing><translation><treatment strategy><type I IFN receptor><type I interferon receptor><vaccination adverse event><vaccination associated adverse event><vaccination induced adverse event><vaccination related adverse event><vaccine adverse event><vaccine against yellow fever><vaccine associated adverse event><vaccine development><vaccine induced adverse event><vaccine related adverse event><viral RNA><viral infection><viral multiplication><viral replication><virus RNA><virus infection><virus multiplication><virus protein><virus-induced disease><yellow fever virus vaccine>