Regulation of the Unfolded Protein Response after Acute Brain Injury

Project: Research project

Project Details

Description

The proposed research investigates novel molecular targets and processes that promise to minimize brain
damage after cerebral ischemic stroke (CIS) and traumatic brain injury (TBI). Current therapeutic strategies
to combat acute brain injuries have been largely unsuccessful. We discovered that the Ca2+ dependent
phosphatase calcineurin (CN) can bind to PERK, a stress sensor in the endoplasmic reticulum (ER),
increases its auto-phosphorylation and enhance a cellular process called the Unfolded Protein Response
(UPR). The UPR attenuates protein translation during stress and gives the cell more time to recover.
Significantly, our preliminary data suggest that this new protective role for CN increases cell viability after
ischemic conditions in cell culture. The goal of this R21 proposal is to develop molecular interventions that
can be used to specifically regulate PERK auto-phosphorylation in vivo. Ultimately, data generated from this
proposal will be used to delineate the therapeutic potential of regulating the UPR during CIS and TBI. Our
overall hypothesis is that, under ischemic conditions, CN directly interacts with PERK with Ca2+ as a co-factor.
The formation of this protein complex promotes PERK dimerization/oligomerization and auto-phosphorylation.
This, in turn, enhances inhibition of protein translation and cell viability, which reduces brain damage after
injury. We have two Specific Aims: 1) Develop molecular interventions that promote, disrupt or mimic CN
binding to PERK. 2) Delineate the Ca2+ dependence of CN binding to PERK in vivo.
Biochemical assays and biophysical techniques will be used to map the binding interaction of CN and PERK
and to generate the peptide fragments. Primary cultures of astrocytes will be used to test the efficacy of
these molecular interventions in vivo. Confocal microscopy will be used to image changes in microdomains
of Ca2+ near the ER. Oxygen Glucose Deprivation, an in vitro model of ischemia, will be used to determine
the physiological impact of PERK auto-phosphorylation as well as our molecular interventions. If successful,
the development of these peptides will serve as attractive therapeutic tools for the treatment of brain injuries.
StatusFinished
Effective start/end date9/29/138/31/16

Funding

  • National Institutes of Health: $164,463.00
  • National Institutes of Health: $205,020.00

ASJC

  • Medicine(all)
  • Neuroscience(all)

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