# Project 2

> **NIH NIH P01** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2022 · $504,146

## Abstract

Project 2 explores the paradigm-shifting hypothesis that the epithelium is at the heart of two
fundamentally different molecular phenotypes of severe asthma (SA). In the 1st, bronchial
epithelial cells (BEC)s respond to initial environmental stimuli to drive an innate intrinsic
phenotype. In the 2nd, BEC cell death pathways intersect with CD8 T-cell immune processes to
drive an immune interactive phenotype. This hypothesis is based on published, submitted and
preliminary data across Project 1 and Project 2 which identify two immune cell phenotypes of
severe asthma, one independent of T-cells and another, highly associated with them, in particular
with CD8 cells. Similar levels of Type-2 (T2) biomarkers/genes are present in both. In published
data from the previous cycle, epithelial growth and repair modules more strongly associated with
asthma severity than T2 modules, suggesting T2 immunity alone does not drive severity.
Consistent with gene expression, poor wound repair of asthmatic BECs in vitro was reported,
with associated loss of a proliferative capacity. The tethered mucin MUC4b, increased in asthma,
lowers BEC proliferation, while increasing secretion. BEC gene clustering mirrors Project 1 with
T-cell- and non-T-cell associated SA phenotypes, each with similar levels of T2 biomarkers. The
innate intrinsic/Non-T-cell SA cluster associates with innate NF-kB and apoptotic pathways
potentially through Toll like receptors (TLR)s. The immune interactive/CD8 cluster associates
with IL-12 and -18, interferons and upregulation of inflammasome and pyroptosis pathways
through the 17q12-21 gene, GSDMB. Thus, interactions between BEC genetic/epigenetic risks,
innate and adaptive immunity and cell death pathways may contribute to these distinct
phenotypes, with differing treatment responses. Using cutting edge technologies of scRNAseq,
advanced bioinformatics and mechanistic in vitro and in vivo models, we will refine the molecular
phenotypes identified in the 1st cycle and link them with cell profiles identified in Project 1. We
propose 3 aims: 1) Identify the mechanisms for and functional implications of an innate intrinsic
epithelial phenotype using fresh and cultured healthy and asthmatic BECs 2) Evaluate the role
of GSDMB in the development of a IL-18/CD8-associated immune interactive phenotype and
its functional implications using fresh and cultured healthy and asthmatic BECs 3) Integrate
immune-inflammatory phenotypes with epithelial molecular phenotypes, as the penultimate effort
to link cell types, mechanisms and compartments to rigorously refine phenotypes. These studies
will improve the understanding of molecular phenotypes in relation to cell death, allowing
Identification of novel precision therapeutic targets to eventually confirm underlying endotypes.

## Key facts

- **NIH application ID:** 10425158
- **Project number:** 2P01AI106684-06A1
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Sally E Wenzel
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $504,146
- **Award type:** 2
- **Project period:** 2015-06-01 → 2027-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10425158

## Citation

> US National Institutes of Health, RePORTER application 10425158, Project 2 (2P01AI106684-06A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10425158. Licensed CC0.

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