# Role of Cytosolic DNA-multiprotein Interactome in Allergic Airway Inflammation

> **NIH NIH R01** · UNIVERSITY OF TEXAS MED BR GALVESTON · 2020 · $790,695

## Abstract

PROJECT SUMMARY/ABSTRACT
Allergic rhinitis and asthma are highly prevalent diseases characterized by allergic airway inflammation. The
airway epithelium is the first line of defense against allergens. We reported that allergenic extracts stimulate
oxidative DNA damage and innate immune responses in these cells. Here we show that cytosolic double
strand (ds)DNA forms a complex with several proteins in the nasal mucosa of humans with allergic rhinitis and
lungs of allergic mice, but not in non-sensitized humans and mice. We named this cytoplasmic DNA-
interactome “Allergosome”. The DNA-interacting proteins in the Allergosome are: interferon regulatory factor 3
(IRF3), the DNA glycosylase Nei-Like 2 (Neil2), and Cyclic GMP-AMP synthase (cGAS). Notably, these factors
associate with Stimulator of Interferon Genes (STING), TANK-binding kinase-1 (TBK1), and the pro-allergic
cytokine IL-33 in the Allergosome. The formation and role of the Allergosome in allergic inflammation are gaps
in knowledge that we address in this proposal. The central hypothesis of this proposal is that “Allergic humans
and mice develop an Allergosome in the cytoplasm of the airway mucosa and lungs that stimulates allergic
airway inflammation”. In Aim 1 we will test the hypothesis that human subjects with allergic rhinitis, but not
healthy control subjects, develop an Allergosome with sequestered IL-33 in nasal mucosal cells. Cytosolic and
crosslinked nuclear extracts will be prepared from the nasal punch biopsies of ragweed-allergic and healthy
subjects. The cytosolic extract of the nasal biopsies will be subjected to immunoprecipitation (IP) with anti-IRF3
antibody, and analyzed for associated DNA sequences by Chip-seq, and for associated proteins in the
Allergosome. In the second Aim, we will test the hypothesis that sequestration of Neil2 in the Allergosome
reduces its ability to protect promoter sites in the genome from binding NFκB and stimulating allergic airway
inflammation. Cytosolic and nuclear extracts will be prepared as in Aim 1. The cytosolic extracts will be
subjected to IP with anti-Neil2 antibody, and analyzed as in Aim 1. The nuclear extracts will be subjected to
ChIP-Seq of the IP’d DNA with anti-Neil2 and anti-NFκB antibodies to determine their promoter occupancy,
and gel shift analysis to detect NFκB binding. Recombinant WT and catalytically inactive Neil2 will be delivered
to the airways of sensitized WT mice by engineered peptide carriers, and allergic inflammation will be
reassessed. In Aim 3, we will test the hypothesis that STING and cGAS stabilize the Allergosome and
stimulate allergic airway inflammation. Allergosome formation and CDE-induced allergic inflammation in WT
mice will be compared to that of StingKO mice, lung epithelium-specific inducible cGasKO mice, and WT mice
treated with pharmacologic cGAS inhibitors. These studies will define, for the first time, the central role of the
Allergosome in allergic airway inflammation. We will provide proof...

## Key facts

- **NIH application ID:** 9849331
- **Project number:** 5R01HL145477-02
- **Recipient organization:** UNIVERSITY OF TEXAS MED BR GALVESTON
- **Principal Investigator:** TAPAS K HAZRA
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $790,695
- **Award type:** 5
- **Project period:** 2019-01-10 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9849331

## Citation

> US National Institutes of Health, RePORTER application 9849331, Role of Cytosolic DNA-multiprotein Interactome in Allergic Airway Inflammation (5R01HL145477-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9849331. Licensed CC0.

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