Document text
Principal Investigator: Paul Stephen Keim
Organization: NORTHERN ARIZONA UNIVERSITY
Fiscal Year: 2022
Award: $1,519,099
Funding agency: National Institute of Allergy and Infectious Diseases
OVERALL Section
Title: Early in vivo expressed antigens and their role in virulence, immune response, and vaccines for
coccidioidomycosis.
SUMMARY
Coccidioidomycosis, also known as Valley Fever (VF), is an important fungal disease caused by two different
Coccidioides species that results in regionally important mortality and even greater morbidity. We have
assembled a team to define the changes that occur in human and animal immune responses to VF, and to use
this knowledge for designing new vaccines and diagnostic tests. This work will capitalize upon our detailed
observations of Coccidioides gene expression patterns during the earliest stages of infections. We hypothesize
that some Coccidioides early genes are virulence factors and critical for causing disease. Research Project 1
will test their role through gene knockouts using CRISPR-Cas9 technology and virulence testing in wax worm
(Galleria) and mouse VF models. Critical virulence factors will become diagnostic and vaccine targets. In addition
to the wax worm and mouse models, we develop a non-human primate (pig-tailed macaques) that will more
closely resemble VF in humans. In humans and vertebrate animal models, the role of T cells cannot be
overemphasized and Research Project 2 will use focused deep DNA sequencing to identify classes of T cell
receptors (TCR) that develop in response to early expressed Coccidioides genes. We will generate TCR
sequences from patients at three clinical locations that span the endemic zones for the pathogen. The TCR
repertoire from patients will be used to identify novel diagnostic signatures (e.g., public TCRs) and, also, help
identify immune responses to key antigens that can be targeted for vaccine development. Hence, both TCR and
early virulence genes represent excellent candidates for vaccine design that will be explored in Research
Project 3 using nucleic acid (NA) based vaccines (RNA and DNA) that can rapidly test a large panel of antigens
through the immunization of mice against infection. The DNA vaccine will be based upon delivery on gold
nanoparticles and Gene Gun, while the mRNA employs self-replicating RNA molecules (repRNA) and a Lipid
InOrganic Nanoparticle (LION). Both are proven technologies that are moving forward into clinical trials for other
diseases. Our goal in the NA vaccine mouse studies is to identify the best antigens and delivery modality for
vaccine testing in the NHP model and to define their immune mechanisms of protection. This work is only
possible through the integrated efforts of investigators at seven different institutions, including three clinical sites,
as no single institution has the requisite breadth of expertise and infrastructure. While we will generate
fundamental knowledge about Coccidioides and VF, we will also make translational advances towards
preventing and diagnosing the disease.
Terms: <Animal Model><Animal Models and Related Studies><Animals><Antibodies><Antigenic Determinants><Antigens><Basic Research><Basic Science><Binding Determinants><Biolistics><C immitis><C posadasii><C. immitis><C. posadasii><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Clinical><Clinical Trials><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><Coccidioides><Coccidioides immitis><Coccidioides posadasii><Coccidioidomycosis><Communities><Complex><DNA><DNA Vaccines><DNA seq><DNA sequencing><DNAseq><Data><Deoxyribonucleic Acid><Desert rheumatism><Development><Diagnostic><Diagnostic tests><Disease><Disorder><Epitopes><Expression Signature><Foundations><Gene Expression><Gene Expression Profile><Gene-Gun Technique><Genes><Goals><Human><Immune><Immune response><Immunes><Immunization><Immunologic Sensitization><Immunologic Stimulation><Immunological Sensitization><Immunological Stimulation><Immunological response><Immunostimulation><Infection><Infrastructure><Insecta><Insects><Insects Invertebrates><Institution><Investigators><Knock-out><Knockout><Knowledge><Link><Lipids><Location><M nemestrina><M. nemestrina><MHC Receptor><Macaca nemestrina><Major Histocompatibility Complex Receptor><Messenger RNA><Mice><Mice Mammals><Modality><Modeling><Modern Man><Morbidity><Morbidity - disease rate><Murine><Mus><Naked DNA Vaccines><Non-Polyadenylated RNA><Nucleic Acid Amplification Tests><Nucleic Acid Testing><Nucleic Acid Vaccines><Nucleic Acids><Pathogenesis><Pathogenicity Factors><Patients><Pattern><Peptides><Pigtail Macaque><Pigtail Monkey><Population><Prevention><Process><Proteins><R-Series Research Projects><R01 Mechanism><R01 Program><RNA><RNA Gene Products><RNA Seq><RNA immunization><RNA sequencing><RNA vaccination><RNA vaccine><RNA-based vaccine><RNAseq><Recombinant DNA Vaccines><Research><Research Grants><Research Personnel><Research Project Grants><Research Projects><Researchers><Ribonucleic Acid><Role><San Joaquin Valley fever><Serology test><T cell response><T-Cell Antigen Receptors><T-Cell Receptor><T-Cells><T-Lymphocyte><T-cell receptor repertoire><TCR repertoire><Technology><Testing><Therapeutic><Therapeutic Agents><Time><Translational Research><Translational Science><United States><Vaccination><Vaccine Antigen><Vaccine Design><Vaccines><Valley Fever><Vertebrate Animals><Vertebrates><Virulence><Virulence Factors><Waxes><Work><base><clinical research site><clinical site><companion diagnostics><desert fever><design><designing><develop a vaccine><develop vaccines><development of a vaccine><developmental><diagnostic approach><diagnostic profile><diagnostic signature><diagnostic strategy><diagnostic tool><disease diagnosis><evaluate vaccines><gene expression pattern><gene expression signature><gene gun><gene product><gold nano particle><gold nanoparticle><host response><human disease><immune system response><immunogen><immunoresponse><in vivo><insight><knockout gene><longitudinal analysis><mRNA><mRNA immunization><mRNA vaccination><mRNA vaccine><mRNA-based vaccine><model of animal><model organism><mortality><mouse model><murine model><nano gold><nano particle><nano-sized particle><nano-string><nanoGold><nanoparticle><nanosized particle><nanostring><new diagnostics><new vaccines><next generation diagnostics><next generation vaccines><non-human primate><nonhuman primate><novel><novel diagnostics><novel vaccines><nucleic acid-based vaccine><pathogen><pig-tailed macaque><prevent><preventing><respiratory><response><screening><serological marker><serology assay><social role><therapeutic evaluation><therapeutic testing><thymus derived lymphocyte><tool><transcriptional profile><transcriptional signature><transcriptome sequencing><translation research><vaccine candidate><vaccine development><vaccine evaluation><vaccine screening><vaccine testing><vertebrata>