Imaging of Functionally Distinct Leukocyte Subtypes within Tissue Biopsies Using Gene Expression Profiling and a Multiple RNA Targeting Approach Imaging of Functionally Distinct Leukocyte Subtypes within Tissue Biopsies Using Gene Expression Profiling and a Multiple RNA Targeting Approach Allergic diseases are characterized by the local tissue infiltration of unique inflammatory leukocytes that are often dominated by eosinophils. These infiltrating eosinophil are currently perceived as a single monolithic population, although recent studies suggest that unique eosinophil subtypes that potentially display distinct activities co-exist. Unfortunately, efforts to stratify eosinophils into functionally distinct groups has thus far proven to be difficult and time consuming, leaving the role of individual cells largely unknown. This proposal integrates growing definitions of eosinophil subtype-specific gene expression with a novel multi-RNA targeting approach, allowing single cell gene expression imaging within tissue biopsies by reiterative cycles of in situ hybridization. The central hypothesis of these studies are that the immune polarization of tissue infiltrating eosinophils is disease specific, diagnostic of ongoing local immune responses, and potentially a prognostic indicator of disease outcomes. Our immediate objectives are to exploit the ongoing studies defining eosinophil subtypes based on gene expression profiling and then use this multi-RNA targeting approach to image unique subtypes in biopsies from established mouse models of asthma. The long term goal beyond this proposal is to translate this strategic approach to human subjects by developing signature eosinophil gene expression biomarkers that will allow physicians to image patient biopsies as part of novel diagnostic approaches to treat allergic diseases. The objectives will be accomplished by the completion of the following Specific Aims: (1) To develop an experimental strategy based on a multiple RNA targeting approach using cleavable fluorescent probes and reiterative in situ imaging. Our objective is to define and localize functionally distinct immune polarized eosinophil subtypes infiltrating the lung following allergen provocation via a signature pattern of gene expression. (2) To define the stratification of immune polarized eosinophil subtypes resident in tissues at homeostatic baseline in mice (e.g., the gastrointestinal tract and uterus) as well as unique disease settings (e.g., solid tumors, helminth infection, allergic skin inflammation) using our multi- RNA targeting approach.
|Effective start/end date||2/6/17 → 6/30/18|
- Mayo Clinic Arizona: $25,000.00
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