Associate Investigator,
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I am interested in studying the molecular and cellular basis of neurological
diseases, especially in ischemic stroke. Stroke is a potentially devastating
neurological disorder with high morbidity and mortality. Thrombolytic agents
such as tissue plasminogen activator (t-PA) are currently the only drugs
available to reverse acute ischemic stroke, but reestablishment of circulation
may paradoxically initiate a reperfusion injury. Therefore, there is great
interest in developing treatments to limit reperfusion injury. In collaboration
with Drs. Donna Ferriero and Clifford Lowell at UCSF, we found that PKCδ null
mice show a striking 70% reduction in stroke size compared with wild type mice
after transient middle cerebral artery (MCA) occlusion and reperfusion. This was
associated with reduced infiltration of peripheral blood neutrophils into
infarcted tissue, as well as impaired neutrophil adhesion, migration,
respiratory burst, and degranulation in vitro. To confirm whether
impaired neutrophil function was important for the improved stroke phenotype in
PKCδ null mice, we treated mice with total body irradiation followed by
transplantation with bone marrow from the opposite genotype. We found that
transplantation reversed the stroke phenotypes in wild type and PKCδ null mice,
which is consistent with an important role for neutrophil PKCδ in reperfusion
injury. Understanding the molecular and cellular actions of PKCδ in reperfusion
injury requires the identification of the unique targets of PKCδ in signaling
pathways activated by ischemia and reperfusion. A novel chemical-genetics
approach has been developed by Dr. Kevan Shokat at UCSF to identify immediate
substrates of kinases. Based on this approach, we have generated a PKCδ-analog
sensitive mutant (PKCδ-as) enzyme, and recently we succeeded in generating a
knock-in mouse expressing PKCδ-as. With these reagents, we have the unique
opportunity to identify direct substrates of PKCδ in response to
stroke-reperfusion injury. The information obtained will reveal PKCδ signaling
pathways activated during cerebral ischemia and reperfusion in neutrophils, and
should facilitate the development of PKCδ-related therapeutic strategies.
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