Funding tool: Basic research projects
Project-ID: LS18-022
Project start: 01. Dezember 2019
Project end: will follow
Runtime: 36 months / ongoing
Funding amount: € 300.000,00

Brief summary:

Allergic disease is a clinical and societal burden in Lower Austria and elsewhere. Severe food allergy can manifest as anaphylactic reaction with potentially fatal outcome. IgE-mediated food allergy in children is increasing, affecting 3-8% of under 4 year olds, with peanut as most common specificity. IgE-producing B-cells and IgE-primed mast cells are critical to TH2 driven, Type I hypersensitivity allergic reactions, but the full scope of altered immunity in these patients and the basis for oral immunotherapy efficacy are unknown, particularly in the solid tissues of the human body.
Here, we propose to analyze the cellular organization of the human immune system in health and allergic disease. Our interdisciplinary consortium consists of Dr. Danzl (IST) providing optical imaging, Dr. Boyd (Stanford) contributing immunology expertise, and clinical partners Dr. Maieron and Dr. Sprinzl (St. Pölten) providing human patient tissue biopsies to link our scientific findings to the clinic.
We will further develop imaging technology to characterize transcriptional profiles of single cells in native tissue context. This identifies cell types, subtypes, and activation states, as well as their spatial arrangements and interactions. We will define molecularly-informed cellular microenvironments or “tissue niches” by detecting hundreds to thousands of different mRNAs in highly multiplex single-molecule RNA fluorescence in situ hybridization. We will also develop multiplex protein imaging, with the same goal of decoding patterns of spatial relationships.
We will verify detection of known spatial organization features in human tonsils and Peyer’s patches. We will then extract novel information by defining cell type specific “interactomes” and specific microenvironments or niches based on detailed molecular and spatial information in our multiplex imaging.
We then focus on gastrointestinal (GI) mucosa-associated immune cells and characterize microenvironments of lymphocytes and effector cells. We emphasize IgE-producing B-cells/plasma cells, and evaluate how their microenvironments differ from those of B-cells and plasma cells producing other antibody isotypes, such as IgA or IgG4. The latter may serve a protective role against allergic disease.
We translate these scientific questions to the clinic by multiplex analysis of patient GI biopsies from a Stanford peanut allergy immunotherapy trial. We hypothesize that not only abundance and location of specific isotype-producing B-cells are shifted in allergy but that specific microenvironments act as major drivers of disease and therapeutic responses. Multiplex single cell analysis of GI biopsies of the same patients pre and post oral immunotherapy will identify therapy-related changes in cellular composition, phenotype, and tissue architecture. We propose to define multi-parameter, spatially informed biopsy biomarkers predictive of treatment response as basis for a personalized medicine approach to immunotherapy.

Keywords:
Optical imaging, microscopy, super-resolution imaging, single-cell analysis, pathology, food allergy