# Mechanisms of Peripheral Remodeling of the EC Cytoskeleton

> **NIH NIH P01** · UNIVERSITY OF ARIZONA · 2020 · $374,726

## Abstract

ABSTRACT:
The endothelial cell (EC) cytoskeleton is a complex array of proteins intimately involved in the diverse cell
shape changes critical to dynamic regulation of lung vascular barrier integrity, particularly in the pathobiology of
and recovery from ARDS and VILI. Consistent with the thematic underpinnings of this PPG, Project #2
investigators have identified specific cytoskeletal effectors (cortactin, Ena/VASP-like protein or EVL, c-Abl)
involved in the actin-based cytoskeletal remodeling that is spatially defined at the EC periphery and within
lamellipodia protruding into the paracellular space. Complementing studies described in Project #1, Specific
Aim #1 will focus on Arp2/3-mediated actin polymerization dynamics in vitro and the effects of c-Abl-mediated
post-translational modifications (PTMs, tyrosine phosphorylation) and cytoskeletal effector coding
polymorphisms (SNPs) on rates of actin polymerization. Specific Aim #2 will next define the critical
participation of key cytoskeletal effector proteins (cortactin, EVL, c-Abl) in regulation of peripheral actin
cytoskeletal remodeling, and EC barrier responses (lamellipodia formation, paracellular gap closure, barrier
restoration). In addition, the functional contribution of specific cytoskeletal effector PTMs, including c-Abl-
mediated cytoskeletal protein tyrosine phosphorylation, will be assessed. These studies will employ Core D
expertise and utilize mutant fusion proteins, live cell imaging with kymography, super resolution, atomic and
traction force microscopy, protein-protein binding assays, and intercellular gap closure assays to visualize the
contributions of these peripheral cytoskeletal effector proteins to EC barrier responses. Similarly, Project #2
investigators have preliminarily defined key non-synonymous coding SNPs that significantly influence protein
functionality in peripheral remodeling and EC barrier restoration. Specific Aim #3 will interrogate the influence
of cytoskeletal effector coding SNPs that we speculate affect cytoskeletal effector protein function either via
altered protein-protein interactions (cortactin-EVL interaction) or altered kinase activity (c-Abl) within the
peripheral actin network. These studies, designed to define the functional consequences of specific coding
SNPs within each cytoskeletal effector protein, will use the array of biophysical and imaging modalities
described for SA #2. Finally, utilizing preclinical models of ARDS and VILI (Core C), Specific Aim #4 will
characterize the in vivo influence of cytoskeletal effector protein PTMs and SNPs that we have determined to
be impactful in vitro. Based on our compelling published and preliminary data that strongly indicate that each
cytoskeletal effector protein to be studied in Project #2 is required to produce agonist-induced lung EC barrier
enhancement, Project #2 cytoskeletal effectors are attractive molecular targets for novel therapies and warrant
intense structure/function investigation.

## Key facts

- **NIH application ID:** 9925260
- **Project number:** 5P01HL126609-05
- **Recipient organization:** UNIVERSITY OF ARIZONA
- **Principal Investigator:** STEVEN M DUDEK
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $374,726
- **Award type:** 5
- **Project period:** — → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9925260

## Citation

> US National Institutes of Health, RePORTER application 9925260, Mechanisms of Peripheral Remodeling of the EC Cytoskeleton (5P01HL126609-05). Retrieved via AI Analytics 2026-06-14 from https://api.ai-analytics.org/grant/nih/9925260. Licensed CC0.

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