# Change in NSF ATPase activity Leads to Brain Ischemia Reperfusion Injury

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA, SAN DIEGO · 2023 · $311,063

## Abstract

Project Summary: Both focal (stroke) and global (cardiac arrest) brain ischemia are major causes of
death and long-term disability, but the underlying mechanisms are still not completely understood. The
objective of this proposal is to study a novel hypothesis that both focal and global brain ischemia lead to a
cascade of events of inactivation of N-ethylmaleimide sensitive factor (NSF), massive buildup of damaged
Golgi-endosomal structures, fatal cathepsin B (CTSB) release, induction of mitochondrial outer membrane
permeabilization (MOMP), and brain ischemia-reperfusion injury (IRI).
 NSF is the sole ATPase for controlling membrane trafficking from Golgi apparatus to the endosome-
lysosome system. Our recent studies show that NSF is trapped into inactive aggregates during the early
period of reperfusion in neurons destined to die after both focal and global brain ischemia. EM studies further
show extensive buildup of damaged Golgi/transport vesicles (Vs) and late endosomes (LEs) in postischemic
neurons. Consequently, CTSB is significantly accumulated over time in and eventually released from
damaged Golgi/Vs/LEs, which is followed by induction of MOMP and neuronal death after ischemia. To
study whether NSF inactivation after brain ischemia leads to massive buildup of damaged Golgi/Vs/LEs
and CTSB release, we generated a new neuron-specific NSF activity-deficient transgenic (tg) mouse line.
The most prominent pathological phenotype of this NSF activity-deficient tg mouse line is massive buildup
of damaged Golgi/Vs/LEs and CTSB release, followed by neuronal death, virtually identical to the events
observed in wildtype (wt) neurons destined to die after both focal and global brain ischemia. Moreover,
induced NSF expression in tg mice protects neurons from IRI. Based on these new discoveries, we propose
to test the novel hypothesis strongly supported by preliminary studies, i.e., brain ischemia leads to NSF
inactivation, massive buildup of Golgi/Vs/LEs, fatal CTSB release, induction of MOMP, and eventually IRI.
We will use both focal and global brain ischemia models, two new tg and one knockout (KO) mouse models,
and several cutting-edge technologies to study the molecular processes.
 Aim 1 will test the novel hypothesis that the NSF inactivation-induced cascade of events of massive
buildup of damaged Golgi/Vs/LEs and fatal CTSB release is a common pathway of neuronal death after
both focal and global ischemia. Aim 2 will use a translational focal ischemia model and CTSB KO mice to
test the novel hypothesis that CTSB release plays a key role in execution of neuronal death via induction of
mitochondrial outer membrane permeabilization (MOMP). Aim 3 will use inducible NSF expression tg mice
to test the hypothesis that postischemic expression of (active) NSF alleviates NSF inactivation-induced
damaging events after focal brain ischemia. These studies will provide novel insights into the neuronal death
mechanisms of focal brain IRI and identify new ther...

## Key facts

- **NIH application ID:** 10748602
- **Project number:** 7R01NS102815-05
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN DIEGO
- **Principal Investigator:** Bingren Hu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $311,063
- **Award type:** 7
- **Project period:** 2022-12-06 → 2024-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10748602

## Citation

> US National Institutes of Health, RePORTER application 10748602, Change in NSF ATPase activity Leads to Brain Ischemia Reperfusion Injury (7R01NS102815-05). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10748602. Licensed CC0.

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