RNA in blood is altered prior to hemorrhagic transformation in ischemic stroke.

OBJECTIVE: Hemorrhagic transformation (HT) is a major complication of ischemic stroke that worsens outcomes and increases mortality. Disruption of the blood-brain barrier is a central feature of HT pathogenesis, and leukocytes may contribute to this process. We sought to determine whether ischemic strokes that develop HT have differences in RNA expression in blood within 3 hours of stroke onset prior to treatment with thrombolytic therapy. METHODS: Stroke patient blood samples were obtained prior to treatment with thrombolysis, and leukocyte RNA was assessed by microarray analysis. Strokes that developed HT (n = 11) were compared to strokes without HT (n = 33) and controls (n = 14). Genes were identified (corrected p < 0.05, fold change ≥|1.2|), and functional analysis was performed. RNA prediction of HT in stroke was evaluated using cross-validation, and in a second stroke cohort (n = 52). RESULTS: Ischemic strokes that developed HT had differential expression of 29 genes in circulating leukocytes prior to treatment with thrombolytic therapy. A panel of 6 genes could predict strokes that later developed HT with 80% sensitivity and 70.2% specificity. Key pathways involved in HT of human stroke are described, including amphiregulin, a growth factor that regulates matrix metalloproteinase-9; a shift in transforming growth factor-ß signaling involving SMAD4, INPP5D, and IRAK3; and a disruption of coagulation factors V and VIII. INTERPRETATION: Identified genes correspond to differences in inflammation and coagulation that may predispose to HT in ischemic stroke. Given the adverse impact of HT on stroke outcomes, further evaluation of the identified genes and pathways is warranted to determine their potential as therapeutic targets to reduce HT and as markers of HT risk.

Prediction of cardioembolic, arterial, and lacunar causes of cryptogenic stroke by gene expression and infarct location.

BACKGROUND AND PURPOSE: The cause of ischemic stroke remains unclear, or cryptogenic, in as many as 35% of patients with stroke. Not knowing the cause of stroke restricts optimal implementation of prevention therapy and limits stroke research. We demonstrate how gene expression profiles in blood can be used in conjunction with a measure of infarct location on neuroimaging to predict a probable cause in cryptogenic stroke. METHODS: The cause of cryptogenic stroke was predicted using previously described profiles of differentially expressed genes characteristic of patients with cardioembolic, arterial, and lacunar stroke. RNA was isolated from peripheral blood of 131 cryptogenic strokes and compared with profiles derived from 149 strokes of known cause. Each sample was run on Affymetrix U133 Plus 2.0 microarrays. Cause of cryptogenic stroke was predicted using gene expression in blood and infarct location. RESULTS: Cryptogenic strokes were predicted to be 58% cardioembolic, 18% arterial, 12% lacunar, and 12% unclear etiology. Cryptogenic stroke of predicted cardioembolic etiology had more prior myocardial infarction and higher CHA(2)DS(2)-VASc scores compared with stroke of predicted arterial etiology. Predicted lacunar strokes had higher systolic and diastolic blood pressures and lower National Institutes of Health Stroke Scale compared with predicted arterial and cardioembolic strokes. Cryptogenic strokes of unclear predicted etiology were less likely to have a prior transient ischemic attack or ischemic stroke. CONCLUSIONS: Gene expression in conjunction with a measure of infarct location can predict a probable cause in cryptogenic strokes. Predicted groups require further evaluation to determine whether relevant clinical, imaging, or therapeutic differences exist for each group.

Ischemic transient neurological events identified by immune response to cerebral ischemia.

BACKGROUND AND PURPOSE: Deciphering whether a transient neurological event (TNE) is of ischemic or nonischemic etiology can be challenging. Ischemia of cerebral tissue elicits an immune response in stroke and transient ischemic attack (TIA). This response, as detected by RNA expressed in immune cells, could potentially distinguish ischemic from nonischemic TNE. METHODS: Analysis of 208 TIAs, ischemic strokes, controls, and TNE was performed. RNA from blood was processed on microarrays. TIAs (n=26) and ischemic strokes (n=94) were compared with controls (n=44) to identify differentially expressed genes (false discovery rate <0.05, fold change ≥1.2). Genes common to TIA and stroke were used predict ischemia in TIA diffusion-weighted imaging-positive/minor stroke (n=17), nonischemic TNE (n=13), and TNE of unclear etiology (n=14). RESULTS: Seventy-four genes expressed in TIA were common to those in ischemic stroke. Functional pathways common to TIA and stroke related to activation of innate and adaptive immune systems, involving granulocytes and B cells. A prediction model using 26 of the 74 ischemia genes distinguished TIA and stroke subjects from control subjects with 89% sensitivity and specificity. In the validation cohort, 17 of 17 TIA diffusion-weighted imaging-positive/minor strokes were predicted to be ischemic, and 10 of 13 nonischemic TNE were predicted to be nonischemic. In TNE of unclear etiology, 71% were predicted to be ischemic. These subjects had higher ABCD(2) scores. CONCLUSIONS: A common molecular response to ischemia in TIA and stroke was identified, relating to activation of innate and adaptive immune systems. TNE of ischemic etiology was identified based on gene profiles that may be of clinical use once validated.

The X-chromosome has a different pattern of gene expression in women compared with men with ischemic stroke.

BACKGROUND AND PURPOSE: Differences in ischemic stroke between men and women have been mainly attributed to hormonal effects. However, sex differences in immune response to ischemia may exist. We hypothesized that differential expression of X-chromosome genes in blood immune cells contribute to differences between men and women with ischemic stroke. METHODS: RNA levels of 683 X-chromosome genes were measured on Affymetrix U133 Plus2.0 microarrays. Blood samples from patients with ischemic stroke were obtained at ≤ 3 hours, 5 hours, and 24 hours (n=61; 183 samples) after onset and compared with control subjects without symptomatic vascular diseases (n=109). Sex difference in X-chromosome gene expression was determined using analysis of covariance (false discovery rate ≤ 0.05, fold change ≥ 1.2). RESULTS: At ≤ 3, 5, and 24 hours after stroke, there were 37, 140, and 61 X-chromosome genes, respectively, that changed in women; and 23, 18, and 31 X-chromosome genes that changed in men. Female-specific genes were associated with post-translational modification, small-molecule biochemistry, and cell-cell signaling. Male-specific genes were associated with cellular movement, development, cell-trafficking, and cell death. Altered sex specific X-chromosome gene expression occurred in 2 genes known to be associated with human stroke, including galactosidase A and IDS, mutations of which result in Fabry disease and Hunter syndrome, respectively. CONCLUSIONS: There are differences in X-chromosome gene expression between men and women with ischemic stroke. Future studies are needed to decipher whether these differences are associated with sexually dimorphic immune response, repair or other mechanisms after stroke, or whether some of them represent risk determinants.

Profiles of lacunar and nonlacunar stroke.

OBJECTIVE: Determining which small deep infarcts (SDIs) are of lacunar, arterial, or cardioembolic etiology is challenging, but important in delivering optimal stroke prevention therapy. We sought to distinguish lacunar from nonlacunar causes of SDIs using a gene expression profile. METHODS: A total of 184 ischemic strokes were analyzed. Lacunar stroke was defined as a lacunar syndrome with infarction <15mm in a region supplied by penetrating arteries. RNA from blood was processed on whole genome microarrays. Genes differentially expressed between lacunar (n = 30) and nonlacunar strokes (n = 86) were identified (false discovery rate ≤ 0.05, fold change >|1.5|) and used to develop a prediction model. The model was evaluated by cross-validation and in a second test cohort (n = 36). The etiology of SDIs of unclear cause (SDIs ≥ 15mm or SDIs with potential embolic source) (n = 32) was predicted using the derived model. RESULTS: A 41-gene profile discriminated lacunar from nonlacunar stroke with >90% sensitivity and specificity. Of the 32 SDIs of unclear cause, 15 were predicted to be lacunar, and 17 were predicted to be nonlacunar. The identified profile represents differences in immune response between lacunar and nonlacunar stroke. INTERPRETATION: Profiles of differentially expressed genes can distinguish lacunar from nonlacunar stroke. SDIs of unclear cause were frequently predicted to be of nonlacunar etiology, suggesting that comprehensive workup of SDIs is important to identify potential cardioembolic and arterial causes. Further study is required to evaluate the gene profile in an independent cohort and determine the clinical and treatment implications of SDIs of predicted nonlacunar etiology.

Signatures of cardioembolic and large-vessel ischemic stroke.

OBJECTIVE: The cause of stroke remains unknown or cryptogenic in many patients. We sought to determine whether gene expression signatures in blood can distinguish between cardioembolic and large-vessel causes of stroke, and whether these profiles can predict stroke etiology in the cryptogenic group. METHODS: A total of 194 samples from 76 acute ischemic stroke patients were analyzed. RNA was isolated from blood and run on Affymetrix U133 Plus2.0 microarrays. Genes that distinguish large-vessel from cardioembolic stroke were determined at 3, 5, and 24 hours following stroke onset. Predictors were evaluated using cross-validation and a separate set of patients with known stroke subtype. The cause of cryptogenic stroke was predicted based on a model developed from strokes of known cause and identified predictors. RESULTS: A 40-gene profile differentiated cardioembolic stroke from large-vessel stroke with >95% sensitivity and specificity. A separate 37-gene profile differentiated cardioembolic stroke due to atrial fibrillation from nonatrial fibrillation causes with >90% sensitivity and specificity. The identified genes elucidate differences in inflammation between stroke subtypes. When applied to patients with cryptogenic stroke, 17% are predicted to be large-vessel and 41% to be cardioembolic stroke. Of the cryptogenic strokes predicted to be cardioembolic, 27% were predicted to have atrial fibrillation. INTERPRETATION: Gene expression signatures distinguish cardioembolic from large-vessel causes of ischemic stroke. These gene profiles may add valuable diagnostic information in the management of patients with stroke of unknown etiology though they need to be validated in future independent, large studies.

Distinctive RNA expression profiles in blood associated with white matter hyperintensities in brain.

BACKGROUND AND PURPOSE: White matter hyperintensities (WMH) are areas of high signal detected by T2 and fluid-attenuated inversion recovery sequences on brain MRI. Although associated with aging, cerebrovascular risk factors, and cognitive impairment, the pathogenesis of WMH remains unclear. Thus, RNA expression was assessed in the blood of individuals with and without extensive WMH to search for evidence of oxidative stress, inflammation, and other abnormalities described in WMH lesions in brain. METHODS: Subjects included 20 with extensive WMH (WMH+), 45% of whom had Alzheimer disease, and 18 with minimal WMH (WMH-), 44% of whom had Alzheimer disease. All subjects were clinically evaluated and underwent quantitative MRI. Total RNA from whole blood was processed on human whole genome Affymetrix HU133 Plus 2.0 microarrays. RNA expression was analyzed using an analysis of covariance. RESULTS: Two hundred forty-one genes were differentially regulated at ± 1.2-fold difference (P < 0.005) in subjects with WMH+ as compared to WMH-, regardless of cognitive status and 50 genes were differentially regulated with ± 1.5-fold difference (P < 0.005). Cluster and principal components analyses showed that the expression profiles for these genes distinguished WMH+ from WMH- subjects. Function analyses suggested that WMH-specific genes were associated with oxidative stress, inflammation, detoxification, and hormone signaling, and included genes associated with oligodendrocyte proliferation, axon repair, long-term potentiation, and neurotransmission. CONCLUSIONS: The unique RNA expression profile in blood associated with WMH is consistent with roles of systemic oxidative stress and inflammation, as well as other potential processes in the pathogenesis or consequences of WMH.

Aspirin plus dipyridamole versus aspirin for prevention of vascular events after stroke or TIA: a meta-analysis.

BACKGROUND AND PURPOSE: This meta-analysis systematically reviewed randomized controlled trials comparing aspirin plus dipyridamole with aspirin alone in patients with stroke and TIA to determine the efficacy of these agents in preventing recurrent cerebral and systemic vascular events. METHODS: We performed separate analyses of the incidences of stroke alone and composite outcome of stroke, myocardial infarction, or vascular death. We also performed two subset analyses, planned a priori, to examine effect size based on trials using (1) exclusively immediate-release and (2) predominantly extended-release dipyridamole. RESULTS: The summary results indicate a significant reduction in the overall risk ratio in favor of aspirin plus dipyridamole for stroke alone with relative risk 0.77 (0.67 to 0.89) and the composite end point with relative risk 0.85 (0.76 to 0.94). Studies using immediate-release dipyridamole showed a nonstatistically significant trend in favor of the combination for stroke alone with relative risk 0.83 (0.59 to 1.15) and for the composite outcome with relative risk 0.95 (0.75 to 1.19). Studies using predominantly extended-release dipyridamole showed a statistically significant difference in favor of the combination for stroke alone with relative risk 0.76 (0.65 to 0.89) and for the composite outcome with relative risk 0.82 (0.73 to 0.92). CONCLUSIONS: The combination of aspirin plus dipyridamole is more effective than aspirin alone in preventing stroke and other serious vascular events in patients with minor stroke and TIAs. The risk reduction was greater and statistically significant for studies using primarily extended release dipyridamole, which may reflect a true pharmacological effect or lack of statistical power in studies using immediate release dipyridamole.

Genomic profiles of stroke in blood.

These studies show that gene expression changes in most patients by 2 to 3 hours after ischemic stroke, and in all patients studied by 24 hours.

Clinical application of CT angiography in acute ischemic stroke.

OBJECTIVE: To evaluate the contribution of CT angiography (CTA) in predicting clinical outcome in a broad spectrum of patients presenting with acute neurological deficits suggestive of brain ischemia, to assess its strengths and limitations in this setting, and examine its influence on selection of patients for thrombolytic treatment. PATIENTS AND METHODS: Prospective, observational outcome study of 54 consecutive patients with acute neurological deficits suggestive of brain ischemia who received immediate CTA. Clinical outcome was compared for patients presenting with and without arterial occlusion on CTA. Treatment decisions made by a vascular neurologist blinded to CTA results were compared to CTA cognizant treatment. RESULTS: For patients presenting with slight to moderate neurological deficits, the sensitivity and specificity for predicting good clinical outcome was 0.62 and 0.79, respectively, using the initial NIH Stroke Scale (NIHSS) score alone, and 0.38 and 0.92 if additionally, CTA showed no occlusion. For patients presenting with more severe deficits, the sensitivity and specificity for predicting poor clinical outcome using the NIHSS score alone was 0.79 and 0.60, compared to 0.67 and 0.92 if CTA showed vessel obstruction. CTA correctly identified six stroke mimickers. Selection of patients for thrombolysis made with knowledge of CTA results were more often conservative, and corresponded to CTA blinded decisions in 42/50 cases (84%, r=0.72). CONCLUSIONS: Combining CTA results with the neurological exam allows increased specificity for predicting clinical outcome as compared to predictions based on admission NIH Stroke Scale score alone. Awareness of CTA results was occasionally associated with less aggressive treatment and testing decisions.

Gene expression in blood changes rapidly in neutrophils and monocytes after ischemic stroke in humans: a microarray study.

Ischemic brain and peripheral white blood cells release cytokines, chemokines and other molecules that activate the peripheral white blood cells after stroke. To assess gene expression in these peripheral white blood cells, whole blood was examined using oligonucleotide microarrays in 15 patients at 2.4+/-0.5, 5 and 24 h after onset of ischemic stroke and compared with control blood samples. The 2.4-h blood samples were drawn before patients were treated either with tissue-type plasminogen activator (tPA) alone or with tPA plus Eptifibatide (the Combination approach to Lysis utilizing Eptifibatide And Recombinant tPA trial). Most genes induced in whole blood at 2 to 3 h were also induced at 5 and 24 h. Separate studies showed that the genes induced at 2 to 24 h after stroke were expressed mainly by polymorphonuclear leukocytes and to a lesser degree by monocytes. These genes included: matrix metalloproteinase 9; S100 calcium-binding proteins P, A12 and A9; coagulation factor V; arginase I; carbonic anhydrase IV; lymphocyte antigen 96 (cluster of differentiation (CD)96); monocarboxylic acid transporter (6); ets-2 (erythroblastosis virus E26 oncogene homolog 2); homeobox gene Hox 1.11; cytoskeleton-associated protein 4; N-formylpeptide receptor; ribonuclease-2; N-acetylneuraminate pyruvate lyase; BCL6; glycogen phosphorylase. The fold change of these genes varied from 1.6 to 6.8 and these 18 genes correctly classified 10/15 patients at 2.4 h, 13/15 patients at 5 h and 15/15 patients at 24 h after stroke. These data provide insights into the inflammatory responses after stroke in humans, and should be helpful in diagnosis, understanding etiology and pathogenesis, and guiding acute treatment and development of new treatments for stroke.

Antegrade rheolytic thrombectomy and thrombolysis for superior sagittal sinus thrombosis using burr hole access.

Superior sagittal sinus (SSS) thrombosis has high morbidity and mortality, and urgent recanalization is critical for severe cases. Standard endovascular techniques for thrombolysis and thrombectomy use retrograde venous access, an approach that may be unsuccessful in cases with extensive firm clot burden involving the dural sinuses distal to the SSS. An anterior open transcranial approach to the SSS for catheter sheath placement to facilitate antegrade mechanical thrombectomy and thrombolysis of the SSS and more distal sinuses has not been previously described. Here we describe a case in which multiple unsuccessful attempts at retrograde endovascular access were attempted. Thus, a burr hole over the anterior SSS was performed for daily endovascular antegrade procedures using the Angiojet rheolytic catheter device and chemical thrombolysis. Near-complete recanalization of the SSS was achieved with venous outflow via dilated left transverse and left sigmoid sinuses, along with significant collateral flow in multiple cerebral veins.

CT angiography in acute ischemic stroke: preliminary results.

BACKGROUND AND PURPOSE: We sought to evaluate the ability of CT angiography (CTA) to determine vessel occlusion before acute stroke treatment and to predict its impact on patient outcome. METHODS: Consecutive patients with acute focal neurological deficits received immediate brain CTA. Occlusion on CTA was correlated with other neuroimaging studies and clinical outcome. RESULTS: Diagnostic CTA was obtained in 54 patients: catheter angiography (digital subtraction angiography) confirmed the CTA findings in 12 of 14 patients (86%). CTA results were consistent with at least 1 other neuroimaging study in 40 of 50 patients (80%). Patients with occlusion on CTA had significantly worse discharge National Institutes of Health Stroke Scale (NIHSS) score (mean 14.3 versus 4.5, P=0.0023). In multivariate analysis, both CTA-determined presence of occlusion and admission NIHSS score were independent predictors of clinical outcome. CONCLUSIONS: In our study there was good agreement between acute CTA interpretation and subsequent imaging studies. CTA evidence of occlusion correlated strongly and independently with poor clinical outcome. CTA provides relevant data regarding vessel patency in acute stroke, which may be of value in selecting patients for aggressive treatment.

Antegrade rheolytic thrombectomy and thrombolysis for superior sagittal sinus thrombosis using burr hole access.

Superior sagittal sinus (SSS) thrombosis has high morbidity and mortality, and urgent recanalization is critical for severe cases. Standard endovascular techniques for thrombolysis and thrombectomy use retrograde venous access, an approach that may be unsuccessful in cases with extensive firm clot burden involving the dural sinuses distal to the SSS. An anterior open transcranial approach to the SSS for catheter sheath placement to facilitate antegrade mechanical thrombectomy and thrombolysis of the SSS and more distal sinuses has not been previously described. Here we describe a case in which multiple unsuccessful attempts at retrograde endovascular access were attempted. Thus, a burr hole over the anterior SSS was performed for daily endovascular antegrade procedures using the Angiojet rheolytic catheter device and chemical thrombolysis. Near-complete recanalization of the SSS was achieved with venous outflow via dilated left transverse and left sigmoid sinuses, along with significant collateral flow in multiple cerebral veins.

Effects of gender on gene expression in the blood of ischemic stroke patients.

This study examined the effects of gender on RNA expression after ischemic stroke (IS). RNA obtained from blood of IS patients (n=51; 153 samples at < or =3, 5, and 24 hours) and from matched controls (n=52) were processed on Affymetrix microarrays. Analyses of covariance for stroke versus control samples were performed separately for both genders and the regulated genes for females compared with males. In all, 242, 227, and 338 male-specific genes were regulated at < or =3, 5, and 24 hours after IS, respectively, of which 59 were regulated at all time points. Overall, 774, 3,437, and 571 female-specific stroke genes were regulated at < or =3, 5, and 24 hours, respectively, of which 152 were regulated at all time points. Male-specific stroke genes were associated with integrin, integrin-liked kinase, actin, tight junction, Wnt/ß-catenin, RhoA, fibroblast growth factors (FGF), granzyme, and tumor necrosis factor receptor (TNFR)2 signaling. Female-specific stroke genes were associated with p53, high-mobility group box-1, hypoxia inducible factor (HIF)1α, interleukin (IL)1, IL6, IL12, IL18, acute-phase response, T-helper, macrophage, and estrogen signaling. Cell death signaling was overrepresented in both genders, although the molecules and pathways differed. Gender affects gene expression in the blood of IS patients, which likely implies gender differences in immune, inflammatory, and cell death responses to stroke.

Y chromosome gene expression in the blood of male patients with ischemic stroke compared with male controls.

BACKGROUND: Sex is suggested to be an important determinant of ischemic stroke risk factors, etiology, and outcome. However, the basis for this remains unclear. The Y chromosome is unique in males. Genes expressed in males on the Y chromosome that are associated with stroke may be important genetic contributors to the unique features of males with ischemic stroke, which would be helpful for explaining sex differences observed between men and women. OBJECTIVE: We compared Y chromosome gene expression in males with ischemic stroke and male controls. METHODS: Blood samples were obtained from 40 male patients ≤3, 5, and 24 hours after ischemic stroke and from 41 male controls (July 2003-April 2007). RNA was isolated from blood and was processed using Affymetrix Human U133 Plus 2.0 expression arrays (Affymetrix Inc., Santa Clara, California). Y chromosome genes differentially expressed between male patients with stroke and male control subjects were identified using an ANCOVA adjusted for age and batch. A P < 0.05 and a fold change >1.2 were considered significant. RESULTS: Seven genes on the Y chromosome were differentially expressed in males with ischemic stroke compared with controls. Five of these genes (VAMP7, CSF2RA, SPRY3, DHRSX, and PLCXD1) are located on pseudoautosomal regions of the human Y chromosome. The other 2 genes (EIF1AY and DDX3Y) are located on the nonrecombining region of the human Y chromosome. The identified genes were associated with immunology, RNA metabolism, vesicle fusion, and angiogenesis. CONCLUSIONS: Specific genes on the Y chromosome are differentially expressed in blood after ischemic stroke. These genes provide insight into potential molecular contributors to sex differences in ischemic stroke.

Posterior reversible encephalopathy syndrome in the context of phentermine use resulting in intracranial hemorrhage.

INTRODUCTION: Posterior reversible encephalopathy syndrome (PRES) is a brain disorder characterized by clinical symptoms of headache, visual changes, altered mentation, and seizures. On neuroimaging, focal regions of symmetrical hemispheric edema are most commonly seen in the occipital lobes. It has been suggested that associated intracranial hemorrhage is present in approximately 5% to 17% of cases. We report a case of PRES with large bilateral parieto-occipital hemorrhages in the context of phentermine use. To our knowledge, this is the first reported case of phentermine use associated with PRES. CASE REPORT: A 35-year-old woman taking phentermine for weight loss presented with altered mental status and 2 witnessed generalized seizures. She was found to have large bilateral parieto-occipital hemorrhages. The radiographic imaging was consistent with PRES with hemorrhagic conversion. After treatment with anti-epileptic medication, normalization of blood pressure, and discontinuation of phentermine, the patient improved clinically. Five months after the event, the patient had a repeat brain magnetic resonance imaging that showed resolution of the diffuse edema and hemorrhage. CONCLUSIONS: This case of PRES with hemorrhagic conversation is distinctive in the bilateral and extensive nature of the hemorrhage, and, to our knowledge, is the first reported case of phentermine use likely resulting in PRES. Physicians should be aware of the cerebrovascular risks associated with phentermine use.

Prolonged dependence of ischemic penumbra on induced hypertension.

BACKGROUND: Pharmacological elevation of blood pressure in acute stroke due to large artery occlusion may enhance collateral blood flow and salvage penumbral tissue. The duration of viability of human penumbral tissue under such circumstances is unclear. OBJECTIVE: To describe a patient presenting with acute middle cerebral artery occlusion treated with induced hypertension who had prolonged survival of penumbral tissue with subsequent infarct extension after pressor agents were withdrawn. CONCLUSIONS: Enhanced collateral flow by elevation of blood pressure may result in prolonged survival of the ischemic penumbra which remains dependent on persistent use of pressor agents.