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Determinants of oral anaphylaxis to food

口腔食物过敏的决定因素

基本信息

批准号：
10586739
负责人：
Stephanie Caroline Eisenbarth
金额：
$ 80.48万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-07 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10586739
关键词：
Affect Alleles Allergens Allergic Allergy to eggs Allergy to peanuts Anaphylaxis Antibodies Antigens Backcrossings Basophils Blood Bone Marrow C3H/HeJ Mouse C57BL/6 Mouse CRISPR/Cas technology Candidate Disease Gene Cell Count Cell Differentiation process Cell physiology Cellular biology Child Chimera organism Chromosome 8 Chromosome Mapping Complement DNA Sequence Alteration Data Enzymes Epithelial Cells Exhibits FDA approved Food Food Hypersensitivity Genes Genetic Genetic Screening Genetic Transcription Goals Goblet Cells Human IgE In Vitro Individual Ingestion Inherited Intestinal permeability Lead Life Link Maps Mediating Molecular Mouse Strains Mucous Membrane Mus Oral Pathway interactions Patients Permeability Persons Pharmaceutical Preparations Pharmacologic Substance Phenotype Predisposition Process Quantitative Trait Loci Reaction Recording of previous events Regulation Resistance Resolution Risk SNP genotyping Secretory Cell Surface System Testing Work autosome cell type diagnostic biomarker diagnostic strategy experimental study food allergen forward genetics gastrointestinal epithelium genetic resistance genomic locus in vivo intestinal epithelium mast cell microbiota mouse model novel novel therapeutics oral immunotherapy pharmacologic prevent programs resistance gene single-cell RNA sequencing targeted treatment uptake validation studies

项目摘要

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.

项目总结