Project description:Background and Purpose - Circulating microRNAs (miRNAs) are emerging biomarkers for stroke due to their high stability in the bloodstream and association with pathophysiologic conditions. However, the circulating whole-genome miRNAs (miRNome) has not been characterized comprehensively in the acute phase of stroke. Methods - We profiled the circulating miRNome in mouse models of acute ischemic and hemorrhagic stroke by next-generation sequencing (NGS). Stroke models were compared to sham-operated and naïve mice to identify deregulated circulating miRNAs. Top-ranked miRNAs were validated and further characterized by qRT-PCR. Results - We discovered 24 circulating miRNAs with an altered abundance in the circulation 3 hours following ischemia, whereas the circulating miRNome was not altered after intracerebral hemorrhage compared to sham-operated mice. Among the upregulated miRNA in ischemia, the top-listed miR-1264/1298/448 cluster was strongly dependent on reperfusion in different ischemia models. A time course experiment revealed that the miR-1264/1298/448 cluster peaked in the circulation around 3 hours after reperfusion and gradually decreased thereafter. Conclusions - Alteration of the miRNome in the circulation is associated with cerebral ischemia/reperfusion, but not hemorrhage, suggesting a potential to serve as biomarkers for reperfusion in the acute phase. The pathophysiological role of reperfusion-inducible miR-1264/1298/448 cluster, which is located on chromosome X within the introns of the serotonin receptor HTR2C, requires further investigation.
Project description:The roles of mRNA and microRNA (miRNA) are widely known in many diseases including acute ischemic stroke. About 60 % of all human messenger RNAs (mRNAs) are regulated by microRNAs. Integration analysis using mRNA and miRNA are important to elucidate pathogenesis. But the contribution of mRNA and miRNA, especially miRNA targeted mRNA, related with severity of acute ischemic stroke is not remain understood. To clarify the pathway related with the severity of acute ischemic stroke, we examined mRNA and miRNA integration analysis targeted for acute ischemic stroke.
Project description:The roles of mRNA and microRNA (miRNA) are widely known in many diseases including acute ischemic stroke. About 60 % of all human messenger RNAs (mRNAs) are regulated by microRNAs. Integration analysis using mRNA and miRNA are important to elucidate pathogenesis. But the contribution of mRNA and miRNA, especially miRNA targeted mRNA, related with severity of acute ischemic stroke is not remain understood. To clarify the pathway related with the severity of acute ischemic stroke, we examined mRNA and miRNA integration analysis targeted for acute ischemic stroke.
Project description:Stroke remains a major leading cause of death and disability worldwide. Despite continuous advances, the identification of key molecular signatures of ischemic stroke within the hyper-acute phase of the disease is still of primary interest for a real translational research on stroke diagnosis, prognosis and treatment. High-throughput -omics technologies are enabling large-scale studies on stroke pathology at different molecular levels. Data integration resulting from these -omics approaches is becoming crucial to unravel the interactions among all different molecular elements and highly contribute to interpret all findings in a complex biological context. Here, we have used advanced data integration methods for multi-level joint analysis of transcriptomics and proteomics datasets depicted from the mouse brain 2h after cerebral ischemia. By modeling net-like correlation structures, we identified a set of differentially expressed genes and proteins with a relevant association in stroke pathology. The ischemia-induced deregulation of 10 of these inter-correlated elements was successfully verified in a new cohort of ischemic mice and changes in their expression pattern were also evaluated at a later time-point after cerebral ischemia. Of those, CLDN20, GADD45G, RGS2, BAG5 and CTNND2 were highlighted as potential candidates as blood biomarkers of cerebral ischemia and hence they were evaluated in blood samples from ischemic and sham-control mice and in ischemic strokes and patients presenting stroke-mimicking conditions. Our findings indicated that CTNND2 and GADD45G levels in blood within the first hours after ischemic stroke might be potentially useful to discriminate ischemic strokes from mimics and to predict patients’ poor outcome after stroke, respectively. In summary, we have here used for the first time an integrative approach that enabled us to elucidate by means of biostatistical tools key elements of the initial stages of the stroke pathophysiology and highlight new outstanding proteins that might be further considered as blood biomarkers of ischemic stroke.
Project description:The purpose of this project was to elucidate gene expression in the peripheral whole blood of acute ischemic stroke patients to identify a panel of genes for the diagnosis of acute ischemic stroke. Peripheral blood samples were collected in Paxgene Blood RNA tubes from stroke patients who were >18 years of age with MRI diagnosed ischemic stroke and controls who were non-stroke neurologically healthy. The results suggest a panel of genes can be used to diagnose ischemic stroke, and provide information about the biological pathways involved in the response to acute ischemic stroke in humans. Total RNA extracted from whole blood in n=39 ischemic stroke patients compared to n=24 healthy control subjects.
Project description:INTRODUCTION: Accurate ischemic stroke (IS) etiologic diagnosis provides an appropriate secondary prevention strategy and better outcome. Currently, 45% of IS are cryptogenic urging to find novel approaches to enhance diagnostic precision and secondary stroke prevention. OBJETIVE: To study the transcriptomic content of circulating extracellular vesicles (EVs) to delineate the molecular profile and potential biomarkers of IS etiologies. RESULTS:21 consecutive patients with moderate or severe ischemic stroke (IS) and different IS etiologies were included to perform the transcriptomic analysis of plasma EVs comprising: atherothrombotic (AT, n=6), Cardioembolic (CE, n=10), and embolic stroke of undetermined source (ESUS, n=5). The RNASeq analysis of EVs detected clusters of differentially expressed genes (DEGs) related to IS etiologies.
Project description:Stroke remains a major leading cause of death and disability worldwide. Despite continuous advances, the identification of key molecular signatures of ischemic stroke within the hyper-acute phase of the disease is still of primary interest for a real translational research on stroke diagnosis, prognosis and treatment. High-throughput - omics technologies are enabling large-scale studies on stroke pathology at different molecular levels. Data integration resulting from these -omics approaches is becoming crucial to unravel the interactions among all different molecular elements and highly contribute to interpret all findings in a complex biological context. Here, we have used advanced data integration methods for multi-level joint analysis of transcriptomics and proteomics datasets depicted from the mouse brain 2h after cerebral ischemia. By modeling network-like correlation structures, we identified a set of differentially expressed genes and proteins by ischemia with a relevant association in stroke pathology. The ischemia-induced deregulation of 10 of these inter-correlated elements was successfully verified in a new cohort of ischemic mice, and changes in their expression pattern were also evaluated at a later time-point after cerebral ischemia. Of those, CLDN20, GADD45G, RGS2, BAG5 and CTNND2 were highlighted and evaluated as potential blood biomarkers of cerebral ischemia in blood samples from ischemic and sham-control mice and from ischemic strokes and other patients presenting stroke-mimicking conditions. Our findings indicated that CTNND2 and GADD45G levels in blood within the first hours after ischemic stroke might be potentially useful to discriminate ischemic strokes from mimics and to predict patients’ poor outcome after stroke, respectively. In summary, we have here used for the first time an integrative approach to elucidate by means of biostatistical tools key elements of the initial stages of the stroke pathophysiology, highlighting new outstanding proteins that might be further considered as blood biomarkers of ischemic stroke.
Project description:Ischemic stroke is one of the most common causes of death worldwide and a major cause of acquired disability in adults. Buyang Huanwu decoction (BHD), a traditional Chinese medicine prescription, has long been used clinically for neurological recovery after stroke. Its molecular characteristics and related biomarkers for the therapeutic effect in cerebrospinal fluid (CSF) are yet to be explored. In this project, CSF was obtained from acute ischemic stroke induced mice by a middle cerebral ischemic/reperfusion (CI/R) injury. Proteomic and metabolomic analyses of CSF were performed using LC-MS/MS to define the neuroprotective effect of BHD and the related biomarkers.