# New role for (airway) epithelial PKD in anti-viral immunity

> **NIH NIH F31** · UNIVERSITY OF ROCHESTER · 2020 · $22,760

## Abstract

Project Summary/Abstract
Epithelial cells are a critical first line of defense against pathogens such as influenza and strategies to
modulate epithelial cell signaling are therefore an appealing target for novel therapeutics. We have previously
identified protein kinase D (PKD) as a key regulator of bronchial epithelial barrier integrity during viral infection
and have recently identified PKD as a mediator of airway epithelial chemokine secretion. We hypothesize that
stimulation with the double-stranded RNA polyI:C or Influenza A virus (IAV) infection leads to activation of PKD
(specifically the PKD3 isoform) which increases epithelial chemokine release and promotes leukocyte
recruitment. This project aims to determine the mechanism between PKD3 activity and epithelial human
interleukin-8 (IL-8)/ murine chemokine CXCL1 secretion. To test this aim, we will assay PKD3 activity
(substrate phosphorylation) following siRNA-mediated knock-down of potential receptors upstream PKD
activation, and then use that information to inform identification of the adaptor molecule(s) associated with PKD
(using co-immunoprecipitation). We will also assay IL-8/CXCL1 mRNA levels in human and mouse airway
epithelial cell lines following knock-down of PKD isoforms to determine the isoform driving chemokine
production. We will then use chromatin immunoprecipitation to identify PKD-activated transcription factors
involved in IL-8/CXCL1 transcription and confirm activity with luciferase reporter assay. We further aim to
investigate the therapeutic potential of PKD3 reduction/inhibition to attenuate lung injury following IAV infection.
To test this aim, we will infect PKD3 deficient mice (PKD3-/-) with sub-lethal influenza A virus (IAV) and assay
for leukocyte infiltration (via flow cytometry), chemokine secretion (via ELISA and multiplex array), and general
pathology (histology). We will also generate bone marrow chimeric mice to assess the relative contributions of
PKD3 in stromal cells vs. hematopoietic cells to lung injury and neutrophilia. We hypothesize that stimulation
with the double-stranded RNA polyI:C or Influenza A virus (IAV) infection leads to activation of the PKD3
isoform which increases epithelial pro-inflammatory cytokine release and promotes leukocyte recruitment. We
further hypothesize that PKD3 may play a previously unsuspected role in regulating airway epithelial anti-viral
immune responses and that further study of the mechanism of PKD3-mediated cytokine release in the airway
epithelium may lead to new therapeutics in combating viral respiratory infections.

## Key facts

- **NIH application ID:** 9858409
- **Project number:** 5F31HL140795-03
- **Recipient organization:** UNIVERSITY OF ROCHESTER
- **Principal Investigator:** Janelle Marie Veazey
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $22,760
- **Award type:** 5
- **Project period:** 2018-02-01 → 2020-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9858409

## Citation

> US National Institutes of Health, RePORTER application 9858409, New role for (airway) epithelial PKD in anti-viral immunity (5F31HL140795-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9858409. Licensed CC0.

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