# A protective role for endothelial cell tissue nonspecific alkaline phosphatase in ischemic stroke

> **NIH NIH P20** · WEST VIRGINIA UNIVERSITY · 2020 · $266,000

## Abstract

PROJECT SUMMARY
The objective of this application is to determine how tissue nonspecific alkaline phosphatase (TNAP) enzymatic
activity preserves blood-brain barrier (BBB) function in ischemic stroke. Increased BBB permeability and
oxidative stress are two potential mechanisms through which cerebral ischemia and reperfusion injury elicit BBB
dysfunction and subsequent functional deficits in acute ischemic stroke. TNAP is a highly enriched enzyme in
cerebral microvessels whose function in brain microvascular endothelial cells (BMECs) is poorly understood.
Our preliminary data generated in a cellular BBB model of ischemia-reperfusion injury demonstrate that TNAP
activity stimulates a novel mechanism which enhances cAMP-mediated signaling pathways that suppress Rho
kinase (ROCK) activity and preserve BBB function. These intriguing findings led us to propose the novel concept
that, in the face of cerebral ischemic injury, TNAP-ROCK signaling mechanisms integrate systemic signals at
the BBB to sustain cerebral function and protect against post-stroke behavioral deficits. To address this novel
concept, the application will investigate the central hypothesis that protection from ischemic stroke-induced
impairment of BBB function is mediated by brain endothelial cell TNAP and its associated signaling cascades.
The proposed studies will test this hypothesis in a mouse model of experimental ischemic stroke, transient middle
cerebral artery occlusion (tMCAO), and in an ex vivo model of ischemia-reperfusion injury, oxygen-glucose
deprivation (OGD). The experimental design will employ a mouse model with conditional deletion of TNAP in
endothelial cells (VE-cKO) and its wild type littermates to interrogate TNAP’s effects on BBB permeability, short-
term functional recovery, and BMEC signaling pathways post-stroke. Aim 1 will elucidate how TNAP preserves
BBB permeability and promotes barrier function during cerebral ischemia and reperfusion injury. Aim 2 will
interrogate the role of TNAP-ROCK signaling mechanisms in an OGD model of injury. Aim 3 will integrate the
experiments in Aims 1 and 2 to prepare a competitive R01 application that will lead to independence of COBRE
funding. Taken together, the studies in this proposal will uncover novel insights into the mechanisms that link
TNAP enzyme activity to BBB integrity, barrier function, short-term functional recovery in ischemic stroke. The
translational impact of these studies may uncover TNAP and its associated signaling cascades as novel
therapeutic targets in ischemic stroke and other cerebrovascular disorders.

## Key facts

- **NIH application ID:** 10025934
- **Project number:** 2P20GM109098-06A1
- **Recipient organization:** WEST VIRGINIA UNIVERSITY
- **Principal Investigator:** Candice Brown
- **Activity code:** P20 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $266,000
- **Award type:** 2
- **Project period:** 2014-09-08 → 2025-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10025934

## Citation

> US National Institutes of Health, RePORTER application 10025934, A protective role for endothelial cell tissue nonspecific alkaline phosphatase in ischemic stroke (2P20GM109098-06A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10025934. Licensed CC0.

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