# A multimodal delivery and treatment approach for Acute Lung Injury

> **NIH NIH R01** · UNIVERSITY OF ROCHESTER · 2021 · $566,991

## Abstract

Acute lung injury (ALI) and Acute Respiratory Distress Syndrome (ARDS) are common, devastating clinical
syndromes that affect large numbers of (200,000 cases in the US per year) and have approximately 30%
mortality with the current standard of care. We have developed a highly effective treatment for this disease in
mouse and pig models that uses the ubiquitous overexperssion of the β1 subunit of the Na+,K+-ATPase to
increase alveolar fluid clearance from the previously injured lung. Our experiments show that this treatment not
only improves edema resolution (and lung function and survival), but also improves alveolar epithelial/
endothelial barrier function by upregulating tight junction complexes. Highly efficient and safe gene delivery is
carried out using electroporation, the application of brief synchronized square wave electric pulses across the
chest following naked DNA delivery by aspiration. The procedure causes no trauma, no inflammation, no lung
injury, no cardiac dysfunction, and uses less than 0.1 J/kg of energy in 50 kg healthy or septic pigs. We have
had no deaths from transthoracic electroporation at optimal field strengths in over 90 healthy and 60 septic
pigs with ARDS. We have found that MRCKα, a serine/threonine-protein kinase and a downstream effector of
Cdc42 for cytoskeletal reorganization, is activated by β1 overexpression and is needed for the increased
activity/abundance of tight junction proteins caused by β1. We have shown that these two proteins interact,
that the β1 subunit activates MRCKα, that inhibition or genetic silencing of MRCKα in alveolar type I epithelial
cells abrogates the ability of β1 overexpression to increase tight junction abundance and activity in cultured
cells, and that overexpression of MRCKα improves barrier properties in cultured alveolar type I epithelial cells.
While β1 overexpression increases edema clearance and barrier function, we do not know which activity plays
the predominant role in its treatment ability. Further, the identification of MRCKα may provide a new target for
treatment of ALI/ARDS. We have also found that the miRNA miR-181a that has been reported to be
significantly increased in the serum of ARDS patients, targets the 3'UTRs of both the Na+,K+-ATPase β1
subunit (but not any other Na+,K+-ATPase subunit) and MRCKα. Inhibition of this miRNA by transfection of an
antagomer increases expression of both the β1 subunit and MRCKα in cells. Our studies will also test whether
modulation of miR-181a can increase both the Na+,K+-ATPase β1 subunit and MRCKα to aid resolution of lung
injury in mouse ALI/ARDS models. We will use novel cyclic amphipathic peptide nanoparticles for RNA delivery
that we have used successfully in cells and the mouse lung. The aims are to (1) determine whether improved
alveolar fluid clearance is the primary mechanism by which gene transfer of the Na+,K+-ATPase treats
ALI/ARDS; (2) test whether induction of barrier function by gene transfer of MRCKα can mediate ...

## Key facts

- **NIH application ID:** 10138009
- **Project number:** 5R01HL148825-02
- **Recipient organization:** UNIVERSITY OF ROCHESTER
- **Principal Investigator:** David A Dean
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $566,991
- **Award type:** 5
- **Project period:** 2020-04-05 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10138009

## Citation

> US National Institutes of Health, RePORTER application 10138009, A multimodal delivery and treatment approach for Acute Lung Injury (5R01HL148825-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10138009. Licensed CC0.

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