# A Novel Approach to Target Neutrophilic Airway Inflammation and Airway Hyperresponsiveness in Therapy-Resistant (Refractory) Asthma.

> **NIH NIH R01** · CREIGHTON UNIVERSITY · 2024 · $422,893

## Abstract

Airway hyperresponsiveness (AHR) and airway inflammation are hallmarks of asthma. Refractory asthma
manifests with persistent symptoms despite use of high-dose oral corticosteroids and long-acting β2-agonist
bronchodilators and poses a major healthcare challenge. Understanding the mechanisms and developing
strategies to overcome therapeutic resistance poses a significant unmet need. Airway smooth muscle (ASM)
hypercontraction is a key factor of AHR, and T-helper 17 (Th17) cells promote steroid-insensitive neutrophilic
airway inflammation (NAI). We found p63RhoGEF, a RhoA activator, plays crucial roles in refractory asthma.
We also developed CXN-8, a small molecule inhibitor of p63RhoGEF. Objective: To determine the importance
and mechanisms by which p63RhoGEF modulates the asthma diathesis and if CXN-8 inhibits p63RhoGEF to
ameliorate AHR and NAI. Long-term goal: To develop new therapies for refractory asthma. Findings: 1) p63-
RhoGEF is selectively upregulated in asthmatics and plays a critical role in RhoA activation that controls ASM
hypercontraction and Th17 cell differentiation. 2) CXN-8 inhibits ASM contraction and induces relaxation of
airway. 3) CXN-8 ameliorates AHR and NAI in murine asthma models. 4) CXN-8 interacts with p63RhoGEF to
block RhoA activation. Hypothesis: Targeting p63RhoGEF-stimulated ASM hypercontractility and Th17 cell
differentiation with CXN-8 ameliorates AHR and NAI and that CXN-8 therapy represents a novel strategy for
refractory asthma. We will test this hypothesis in vitro and in vivo. Aim 1. To define the mechanism underlying
p63RhoGEF and CXN-8 regulation of AHR. We will use RNAi and inhibitors to determine the pivotal role and
mechanism of p63RhoGEF in hypercontractility of asthmatic human ASM (HASM) cells. We will analyze CXN-
8 regulation of RhoA activity, Ca2+ signaling, contraction/relaxation of β2-agonist-sensitive/insensitive HASM
cells. We will combine RNAi, mutagenesis, and surface plasmon resonance to elucidate the specificity and
mechanism for CXN-8 regulation of p63RhoGEF. Whether p63RhoGEF loss ameliorates AHR and reduces the
effects of CXN-8 will be examined in a house dust mite (HDM)-driven murine model of asthma. Aim 2. To
elucidate the mechanism underlying p63RhoGEF and CXN-8 regulation of NAI. We will examine the effects of
CXN-8 on Th17 cell differentiation. We will silence RhoA or express an active RhoA mutant to establish its
importance in CXN-8 inhibition of Th17 cell differentiation. We will also determine if loss of p63RhoGEF
attenuates RhoA activation and Th17 differentiation in cells and HDM-induced NAI in mice and reduces CXN-8
inhibitory effects in vitro and in vivo. Aim 3. To examine the therapeutic potential of CXN-8 in murine models of
refractory asthma. We will determine if inhaled CXN-8 is an acute and effective bronchodilator in a murine
model of β2-agonist insensitive AHR. We will test lung targeted, long-acting CXN-8 microparticles to alleviate
AHR/NAI with limited systemic sid...

## Key facts

- **NIH application ID:** 10915460
- **Project number:** 5R01HL164593-02
- **Recipient organization:** CREIGHTON UNIVERSITY
- **Principal Investigator:** YAPING TU
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $422,893
- **Award type:** 5
- **Project period:** 2023-08-20 → 2027-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10915460

## Citation

> US National Institutes of Health, RePORTER application 10915460, A Novel Approach to Target Neutrophilic Airway Inflammation and Airway Hyperresponsiveness in Therapy-Resistant (Refractory) Asthma. (5R01HL164593-02). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10915460. Licensed CC0.

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