Mechanisms For p38 MAP Kinase Dependent Gene Induction

Project: Research project

Project Details

Description

DESCRIPTION (provided by applicant):
This application is aimed at identifying inducible transcription factors that
activate fetal hemoglobin (HbF) expression, for the development of novel
therapies for Sickle cell disease and Cooley's anemia. The rationale for this
strategy is based on clinical, genetic and biochemical evidence illustrating
the beneficial effect of HbF in both genetic disorders. Histone deacetylase
inhibitors (HDAIs) are validated HbF inducers but available analogues are not
widely used due to anti-proliferative effects and poor bioavailability. HDAIs
alter DNA-protein interactions in the fetal globin gene promoter, which
indicates initial activation of hitherto unknown transactivator molecules,
which in turn activate fetal globin. Based on this deduction, we initiated
studies with two HDAIs sodium butyrate and trichosatin A and found both to
activate p38 mitogen activating protein kinase (p38 MAPK) and activating
transcription factor 2 (ATF-2). We have confirmed this mechanism of g globin
activation in experiments using a specific activator (anisomycin) and
inhibitor (SB203580) of p38 MAPK. From these findings we have developed a
hypothesis to be tested in four major specific aims, 1) Determine the
mechanism for p38 MAPK activation by histone deacetylase inhibitors. We will
establish whether HDAI activation of p38 MAPK is direct or through reactive
oxygen species in the mitochondria and test novel HDAIs currently in Phase I
clinical trials for g globin inducibility. 2) Define the cis-regulatory
element(s) required for p38 MAPK dependent g gene induction. These studies
will include functional analysis of the g promoter using reporter and
expression vectors. 3) Identify transcription factor(s) required for p38 MAPK
dependent g gene activation. Cognate binding transcription factors for the
elements identified in Aim 2 will be characterized by gel shift analysis and
immunoprecipitation studies. Experiments in this Aim will include validation
that the identified transactivation or co-activator proteins activate
endogenous g globin. 4) Determine the ability of novel HDAIs to induce g gene
activity in vivo in transgenic lines. Novel HDAIs found to be effective in
Aim 1 will be tested in sickle transgenic mice. Successful completion of the
experiments described in the Specific Aims will provide the basis for
developing effective drug or gene therapy protocols for Sickle cell disease
and Cooley's anemia.
StatusFinished
Effective start/end date9/30/015/31/18

Funding

  • National Heart, Lung, and Blood Institute: $374,992.00
  • National Heart, Lung, and Blood Institute: $400,685.00
  • National Heart, Lung, and Blood Institute: $372,500.00
  • National Heart, Lung, and Blood Institute: $414,337.00
  • National Heart, Lung, and Blood Institute: $372,750.00
  • National Heart, Lung, and Blood Institute: $386,532.00
  • National Heart, Lung, and Blood Institute: $388,730.00

ASJC

  • Medicine(all)