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Principal Investigator: OLGA VINOGRADOVA
Organization: UNIVERSITY OF CONNECTICUT STORRS
Fiscal Year: 2024
Award: $368,411
Funding agency: National Institute of Allergy and Infectious Diseases
Abstract
Immune cells protect us from disease by detecting and responding to foreign molecules. Understanding the
molecular basis of this response is critical if we are to generate new therapies for treatment or prevention of
diseases involving immune cells. The objective of our research is to characterize a newly discovered antigen
detecting protein (butyrophilin 3, BTN3) which influences the immune response mediated by gamma delta T
cells. Gamma delta T cells are cytotoxic T cells that respond quickly to foreign threats and serve a variety of
roles, including direct lysis of infected or malignant cells, and as such, their activation holds great promise for
therapeutic manipulation. In contrast to T cells that express the more prevalent alpha beta T cell receptor and
respond to peptide antigens, T cells that express the Vgamma9Vdelta2 T cell receptor respond to small
phosphorous-containing compounds known as phosphoantigens. Butyrophilin 3A1 (BTN3A1) is the receptor for
phosphoantigens and mediates their activation of T cells through unclear mechanisms. We developed a library
of novel synthetic phosphoantigens, as well as a library of butyrophilin constructs and point mutations, both of
which are valuable tools for understanding the underlying biology of butyrophilins. Here, we propose aims that
test the underlying hypothesis that ligand binding to the intracellular domain of BTN3A1 produces conformational
and organizational changes that are required for interaction with counter receptors on T cells. Understanding
how BTN3A1 and the related 3A2, 3A3, and 2A1 isoforms function at the molecular level is important because it
1) will help optimize past and present clinical trials that have examined phosphoantigens and phosphoantigen-
expanded cells as immunotherapies, and 2) will identify new molecular targets or strategies within this complex
for therapeutic manipulation. Our studies in Aim 1 will show a structural basis for how phosphoantigens affect
the full length endogenous BTN3A1 using multiple biophysical and molecular biological approaches. In Aim 2,
we will investigate the function of BTN3A1 in phosphoantigen-induced Vgamma9Vdelta2 T cell lysis of
phosphoantigen containing cells and associated cytokine production. Together, this will allow us to build a
structure-function model of BTN3 with regards to how its domain organization, oligomerization status, protein-
protein interactions, and relationship to BTN2A1 influence its function. This will largely be done in the context of
biological membranes through use of a novel in vitro membrane nanodisc/cryo-EM model system. Our ultimate
goal is to present a clear structural model that demonstrates how phosphoantigen-induced conformational and/or
compositional changes in the BTN3 complex promote effector functions of T cells. This will enable clinical
development of therapies that modulate butyrophilin function to overcome immune checkpoints. These findings
will come at a point when the biological understanding of antigen detection is far from complete, and thus have
the potential to impact the field.
Terms: <Address><Affect><Antigen-Presenting Cells><Antigenic Determinants><Antigens><Assay><Binding><Binding Determinants><Binding Proteins><Binding Sites><Bioassay><Biologic Models><Biological><Biological Assay><Biological Models><Biology><Biophysics><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><Cancers><Cas nuclease technology><Cell Body><Cell Function><Cell Line><Cell Physiology><Cell Process><Cell Protection><Cell Surface Glycoproteins><Cell Surface Receptors><Cell surface><Cell-Mediated Lympholytic Cells><CellLine><Cells><Cellular Function><Cellular Physiology><Cellular Process><Chemicals><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><Co-Immunoprecipitations><Combining Site><Communicable Diseases><Complex><Conflict><Conflict (Psychology)><Confusion><Confusional State><Cryo-electron Microscopy><Cryoelectron Microscopy><Cytolysis><Cytolytic T-Cell><Cytoplasm><Cytoprotection><Cytotoxic T Cell><Cytotoxic T-Lymphocytes><Data><Disease><Disorder><Distant><Drug Design><Drug Precursors><Electron Cryomicroscopy><Epitopes><Exposure to><External Domain><Extracellular Domain><Extracellular Protein><FRET><Family member><Fluorescence Resonance Energy Transfer><Förster Resonance Energy Transfer><Gamma-delta T cells><Genetic><Goals><Human><Immune><Immune Cell Activation><Immune mediated therapy><Immune response><Immunes><Immunoblotting><Immunological response><Immunologically Directed Therapy><Immunomodulation><Immunotherapy><In Vitro><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Interferon Type I><Isoforms><K562 Cells><Length><Libraries><Ligand Binding><Ligand Binding Protein><Ligand Binding Protein Gene><Ligands><Literature><Lysis><MHC Receptor><Major Histocompatibility Complex Receptor><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Mediating><Membrane><Membrane Glycoproteins><Mental Confusion><Model System><Modeling><Modern Man><Molecular><Molecular Configuration><Molecular Conformation><Molecular Interaction><Molecular Stereochemistry><Molecular Target><Mutate><Organizational Change><Peptides><Phosphorous><Phosphorus><Point Mutation><Pro-Drugs><Prodrugs><Production><Protein Binding><Protein Isoforms><Proteins><Reactive Site><Receptor Protein><Receptors, Antigen, T-Cell, alpha-beta><Reporting><Research><Roentgen Rays><Role><Shapes><Strains Cell Lines><Structural Models><Structure><Subcellular Process><Surface Glycoproteins><T cell response><T-Cell Activation><T-Cell Antigen Receptors><T-Cell Receptor><T-Cells><T-Lymphocyte><TcR alpha-beta><TcR αβ><Testing><Therapeutic><Therapeutic Agents><Visualization><Western Blotting><Western Immunoblotting><X-Radiation><X-Ray Radiation><X-ray><Xray><accessory cell><activate T cells><alpha-beta T-Cell Receptor><analytical ultracentrifugation><antigen based detection><antigen detection><biologic><biophysical foundation><biophysical principles><biophysical sciences><bound protein><butyrophilin><cancer cell><clinical development><conformation><conformational><conformational state><conformationally><conformations><crosslink><cryo-EM><cryoEM><cryogenic electron microscopy><cultured cell line><cytokine><cytoprotective><cytotoxic><detect antigen><develop therapy><dimer><disease prevention><disorder prevention><host response><immune activation><immune check point><immune checkpoint><immune modulation><immune regulation><immune system response><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immunecheckpoint><immuno therapy><immunogen><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><immunoresponse><insight><intervention development><killer T cell><malignancy><membrane structure><mutant><nanodisk><neoplasm/cancer><new drug target><new drug treatments><new druggable target><new drugs><new pharmacological therapeutic><new pharmacotherapy target><new therapeutic target><new therapeutics><new therapy><new therapy target><next generation therapeutics><novel><novel drug target><novel drug treatments><novel druggable target><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel pharmacotherapy target><novel therapeutic target><novel therapeutics><novel therapy><novel therapy target><preservation><protein blotting><protein complex><protein protein interaction><receptor><response><small molecule><social role><therapy development><thymus derived lymphocyte><tool><treatment development><αβ T-Cell Receptor><γδ T cells><γδT cells>