Determinants of oral anaphylaxis to food

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Principal Investigator: Stephanie Caroline Eisenbarth
Organization: NORTHWESTERN UNIVERSITY AT CHICAGO
Fiscal Year: 2024
Award: $798,975
Funding agency: National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY
The goal of this proposal is to identify new ways of preventing anaphylaxis in those with food allergy by defining cellular and molecular mechanisms that transport intact food allergens across the gut epithelium. In its most severe form, food allergy can trigger life-threatening anaphylaxis. One factor that can determine risk of anaphylaxis is the integrity of the gut barrier, but there are no treatments to reduce intestinal permeability to food allergens. A major mode of allergen transport across the gut epithelium occurs transcellularly through secretory and goblet cell-associated antigen passages (GAPs). Using mouse models of peanut and egg allergy, we have made the exciting discovery that susceptibility to oral anaphylaxis was genetically determined and associated with an increased number and function of GAPs. Using a forward genetic screen we identified a single chromosomal region that tracks with this phenotype. Moreover, drug treatments targeting goblet cell biology reduced anaphylaxis in vivo. We hypothesize that the genetic regulation of gut goblet cell quantity and transport capability determines susceptibility to anaphylaxis by controlling intestinal permeability to intact food allergens. In Aim 1, using complementary mouse strains described above, we will perform quantitative trait locus (QTL) mapping with SNP genotyping to identify genetic resistance loci for oral anaphylaxis associated with reduced gut permeability. These data will be integrated with scRNA-Seq analyses of intestinal epithelium from anaphylaxis susceptible vs. resistant littermates to identify cell type-specific regulators of allergen transport. Using CRISPR/Cas9 gene editing in mice, we will test the contribution to gut permeability and oral anaphylaxis in vivo of known and novel candidate genes. In Aim 2 we will determine goblet cell-intrinsic vs. goblet cell extrinsic pathways that inhibit allergen transport. Our preliminary data in mice suggests that the number and function of GAPs prominently contribute to susceptibility to oral anaphylaxis. Using bone marrow chimeras and in vitro human or mouse enteroid cultures we will directly test whether transcellular transport of allergens is increased in anaphylaxis susceptible humans or mice and will determine whether these phenotypes are epithelial cell-intrinsic or -extrinsic. In Aim 3 we will perform a targeted screen of FDA approved drugs that could inhibit goblet cell differentiation and/or function for their ability to block fluorescent allergen uptake in vitro and oral anaphylaxis in vivo. Effective compounds will be validated using human gut enteroids from donors with or without food allergy. We have already identified multiple drugs that reduce allergen transport in oral anaphylaxis susceptible mice, highlighting that pharmaceutical blockade of these pathways could potentially prevent anaphylaxis in patients with food allergy. Impact: Identifying genes and cellular pathways regulating food allergen transport by gut epithelia could lead to new ways of preventing anaphylaxis and to diagnostic approaches to more accurately stratify anaphylaxis risk for food-allergic individuals.

Terms: <0-11 years old><Affect><Alleles><Allelomorphs><Allergens><Allergic><Allergic to food><Allergy to eggs><Allergy to food><Allergy to peanuts><Anaphylactic Reaction><Anaphylactic Shock><Anaphylaxis><Antibodies><Antigens><Backcrossings><Basophilic Granulocyte><Basophils><Blood><Blood Basophil><Blood Reticuloendothelial System><Bone Marrow><Bone Marrow Reticuloendothelial System><C3H/HeJ Mouse><C57BL/6 Mouse><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><Candidate Disease Gene><Candidate Gene><Cas nuclease technology><Cell Count><Cell Differentiation><Cell Differentiation process><Cell Function><Cell Number><Cell Physiology><Cell Process><Cellular Function><Cellular Physiology><Cellular Process><Cellular biology><Child><Child Youth><Children (0-21)><Chimera><Chimera organism><Chromosome 8><Chromosome Mapping><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><DNA Alteration><DNA Sequence Alteration><DNA mutation><Data><Drugs><Egg Hypersensitivity><Enzyme Gene><Enzymes><Epithelial Cells><Exhibits><FDA approved><Food><Food Allergy><Food Hypersensitivity><Gene Localization><Gene Mapping><Gene Mapping Genetics><Gene Transcription><Genes><Genetic><Genetic Screening><Genetic Transcription><Genetic mutation><Goals><Goblet Cells><Groundnut Hypersensitivity><Gut Epithelial Permeability><Gut Epithelium><Gut Hyperpermeability><Gut permeability><Hereditary><History><Human><IgE><Immunoglobulin E><In Vitro><Individual><Ingestion><Inherited><Intestinal Epithelial Permeability><Intestinal Hyperpermeability><Intestinal permeability><Lead><Life><Link><Linkage Mapping><Maps><Marrow Mast Cell><Mediating><Medication><Mice><Mice Mammals><Modern Man><Molecular><Mouse Strains><Mucosa><Mucosal Tissue><Mucous Membrane><Murine><Mus><Oral><Pathway interactions><Patients><Pb element><Peanut Hypersensitivity><Permeability><Persons><Pharmaceutical Agent><Pharmaceutical Preparations><Pharmaceuticals><Pharmacologic Substance><Pharmacological Substance><Phenotype><Predisposition><Process><QTL><Quantitative Trait Loci><RNA Expression><Reaction><Recording of previous events><Regulation><Resistance><Resolution><Risk><SNP genotyping><Secretory Cell><Sequence Alteration><Subcellular Process><Surface><Susceptibility><System><Testing><Tissue Basophils><Total Human and Non-Human Gene Mapping><Transcription><Work><allergic to eggs><allergic to peanuts><autosome><cell biology><cell type><cellular differentiation><chimeras><diagnostic approach><diagnostic biomarker><diagnostic marker><diagnostic strategy><drug/agent><egg allergy><experiment><experimental research><experimental study><experiments><food allergen><forward genetics><gastrointestinal epithelium><gene locus><genetic locus><genetic mapping><genetic resistance><genomic alteration><genomic location><genomic locus><heavy metal Pb><heavy metal lead><histories><immunogen><in vivo><ingest><intestinal epithelium><kids><mast cell><mastocyte><microbial consortia><microbial flora><microbiota><microflora><mouse model><multispecies consortia><murine model><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><oral immunotherapy><pathway><peanut allergy><pharmaceutical><pharmacologic><prevent><preventing><programs><resistance gene><resistance locus><resistant><resistant gene><resolutions><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single nucleotide polymorphism genotyping><single-cell RNA sequencing><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><uptake><validation studies><youngster>