This year in the U.S. close to 800,000 people will suffer a stroke, making it the third leading cause of death. A particularly devastating form of this illness, subarachnoid hemorrhage (SAH), represents 25% of life years lost among all forms of stroke due to the relatively young age of its victims. A SAH occurs when large vessels in the brain rupture and bleed into the fluid filled space surrounding the brain. Patients recovering from SAH often develop delayed neurological deficit due to ischemia, killing a significant number, and leaving survivors with permanent disability. The pathogenesis of this injury is not well understood, and even though several promising mechanisms of delayed neurological deficit have been studied in great detail, treatments developed from this work have had limited success in clinical trials. A recently documented phenomenon that correlates with delayed neurological deficit is delayed microvascular spasm (dMVS). Until now, the technology available to document dMVS has only been able to do so at very low resolution. Using a novel in vivo optical imaging modality called optical microangiography (OMAG), we have been able to directly visualize microvessels within the mouse cortex and definitively identify dMVS after SAH. We propose to use neurobehavioral tests, histology, and biochemistry in addition to OMAG to explore a potential therapeutic target to prevent dMVS. Flow in brain microvessels is controlled by cytochrome P450 metabolites of arachidonic acid called epoxyeicosatrienoic acids (EETs). EETs are inactivated by the enzyme soluble epoxide hydrolase (sEH). Our preliminary data show that sEH knockout mice are protected from dMVS after SAH. We will test the hypothesis that SAH induces upregulation of sEH in cerebrovascular endothelium, leading to a decrease in bioavailable EETs and the development of dMVS and neurological deficit.
In Specific Aim 1, we will test whether genetic deletion and pharmacological inhibition of sEH activity prevent dMVS and neurological deficit in mice after SAH. We will further characterize dMVS and neurological deficit after SAH using OMAG, neurobehavioral examination and histology. Finally, we will use transgenic mice with endothelial overexpression of sEH and the EETs biosynthetic enzyme P450 2J2, to confirm the role of endothelial sEH in modulating dMVS after SAH.
In Specific Aim 2, we will measure changes in sEH expression and EETs within brain microvessels after SAH by Western blotting, real-time quantitative PCR, and mass spectrometry (LC-MS/MS). Additionally, we will test whether the upregulation of sEH after SAH is dependent on angiotensin signaling, which is known to be increased following SAH.
This research aims to identify a novel therapeutic target for prevention of dMVS and delayed neurological deficit caused by SAH.
Individuals recovering from subarachnoid hemorrhage, a form of stroke, often live with significant disability due to delayed neurological deficit that arises well after the initial hemorrhage. Delayed microvascular spasm, a constriction of small resistance vessels and capillaries in the brain by an unknown cause, is associated with a significant number of patients who experience delayed neurological deficit. Here the aim is to study how disruption of a microvascular vasodilator pathway could cause delayed microvascular spasm after subarachnoid hemorrhage and subsequently, delayed neurological deficit.
|Siler, Dominic A; Martini, Ross P; Ward, Jonathan P et al. (2015) Protective role of p450 epoxyeicosanoids in subarachnoid hemorrhage. Neurocrit Care 22:306-19|
|Siler, Dominic A; Berlow, Yosef A; Kukino, Ayaka et al. (2015) Soluble Epoxide Hydrolase in Hydrocephalus, Cerebral Edema, and Vascular Inflammation After Subarachnoid Hemorrhage. Stroke 46:1916-22|
|Siler, Dominic A; Gonzalez, Jorge A; Wang, Ruikang K et al. (2014) Intracisternal administration of tissue plasminogen activator improves cerebrospinal fluid flow and cortical perfusion after subarachnoid hemorrhage in mice. Transl Stroke Res 5:227-37|
|Martini, Ross P; Ward, Jonathan; Siler, Dominic A et al. (2014) Genetic variation in soluble epoxide hydrolase: association with outcome after aneurysmal subarachnoid hemorrhage. J Neurosurg 121:1359-66|