753. Neuroprotective Gene Expression with AAV8 for Oxygen-Glycose Deprivation in Organotypic Hippocampal Culture: A Model of Ischemic Stroke

Background: Alzheimer’s disease is mainly characterized by remarkable neurodegeneration in brain areas related to memory formation. This progressive neurodegeneration causes cognitive impairment, changes in behavior, functional disability, and even death. Our group has demonstrated changes in the kallikrein–kinin system (KKS) in Alzheimer’s disease (AD) experimental models, but there is a lack of evidence about the role of the KKS in Alzheimer’s disease. Aim: In order to answer this question, we evaluated the potential of the kinin B2 receptors (BKB2R) to modify AD characteristics, particularly memory impairment, neurodegeneration, and Aβ peptide deposition. Methods: To assess the effects of B2, we used transgenic Alzheimer’s disease mice treated with B2 receptor (B2R) agonists and antagonists, and performed behavioral and biochemical tests. In addition, we performed organotypic hippocampal culture of wild-type (WT) and transgenic (TG) animals, where the density of cytokines, neurotrophin BDNF, activated astrocyte marker S100B, and cell death were analyzed after treatments. Results: Treatment with the B2R agonist preserved the spatial memory of transgenic mice and decreased amyloid plaque deposition. In organotypic hippocampal culture, treatment with B2R agonist decreased cell death, neuroinflammation, and S100B levels, and increased BDNF release. Conclusions: Our results indicate that the kallikrein–kinin system plays a beneficial role in Alzheimer’s disease through B2R activation. The use of B2R agonists could, therefore, be a possible therapeutic option for patients diagnosed with Alzheimer’s disease.

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Obtaining Human Ischemic Stroke Gene Expression Biomarkers from Animal Models: A Cross-species Validation Study

Scientific Reports ◽

10.1038/srep29693 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 7

Author(s):

Yingying Wang ◽

Yunpeng Cai

Keyword(s):

Gene Expression ◽

Ischemic Stroke ◽

Animal Models ◽

Validation Study

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Elevated Lipocalin-2 Can Indicate the Vascular Inflammation in Patients with Ischemic Stroke

Austin Journal of Cerebrovascular Disease & Stroke ◽

10.26420/austinjcerebrovascdisstroke.2019.1081 ◽

2019 ◽

Author(s):

Rajnics P ◽

◽

Kellner A ◽

Nagy F ◽

Alföldi V ◽

...

Keyword(s):

Gene Expression ◽

Ischemic Stroke ◽

Gene Expression Level ◽

Expression Level ◽

Neutrophil Granulocytes ◽

Lipocalin 2 ◽

Relative Gene Expression ◽

Thrombotic Risk ◽

Relative Gene ◽

Relative Gene Expression Level

Purpose: Elevated level of Lipocalin-2 (LCN2), a new acute phase adipokine, was described after ischemic stroke. A number of researchers feel as though that LCN2 originated from the infiltrating neutrophils and other cells in brain after stroke. Others measured elevated LCN2 expression in arteriosclerotic plaque. Therefore we have investigated LCN2 relative gene expression level of blood neutrophil granulocytes in patients with ischemic stroke to assess if elevated LCN2 is the cause or consequence of ischemic stroke. Methods: Laboratory and anamnestic data were collected, which could have a role in development of thrombo-embolic events in patients with ischemic stroke. RNA based method was used to evaluate the relative gene expression level of LCN2. We calculated Odds Ratio (OR) and Confidence Interval (CI) for the association between LCN2 and ischemic stroke. Results: 34 samples were available for evaluation. The LCN 2 relative gene expression level was decreased in 12 cases. In this group, 91% of patients have Atrial Fibrillation (AF) at the time of hospitalisation. The mean LCN2 relative gene expression value was 64.25% (ranges: 34%-115%) in patients with AF. It was significantly lower than in patients with normal sinus rhythm (409.2%; ranges: 127%-1127%; p=0.0003). The elevated LCN2 relative gene expression level significantly (p=0.012) increases the risk of stroke (OR: 12.6) independently from other factors. Conclusions: High LCN2 expression level seems to have strong positive predictive value on ischemic stroke, and may be useful in thrombotic risk stratification of plaque vulnerability in these patients.

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Hyperexpression of TLR2 and TLR4 in patients with ischemic stroke in acute period of the disease

Medical Immunology (Russia) ◽

10.15789/1563-0625-hot-1971 ◽

2020 ◽

Vol 22 (4) ◽

pp. 665-674

Author(s):

L. V. Gankovskaya ◽

L. V. Stakhovskaya ◽

V. V. Grechenko ◽

E. A. Koltsova ◽

O. S. Uvarova ◽

...

Keyword(s):

Gene Expression ◽

Innate Immunity ◽

Ischemic Stroke ◽

Peripheral Blood ◽

Surface Expression ◽

Control Group ◽

Peripheral Blood Leukocytes ◽

Blood Leukocytes ◽

Healthy Donors ◽

Tlr4 Gene

Pathogenesis of ischemic stroke is actively involved in the system of innate immunity. Under conditions of cerebral ischemia, a number of biologically active substances are released that interact with innate immunity receptors, in particular TLR2 and TLR4, which exacerbate inflammation in brain tissue. Identification of predictor markers at the level of the innate immunity system may foresee the clinical course of ischemic stroke and ensure timely treatment. Our objective was to study expression of TLR2 and TLR4 receptors in peripheral blood leukocytes in patients with ischemic stroke in the dynamics of the disease. 27 people were included in the study. The main group consisted of patients with ischemic stroke of varying severity (n = 19). Patients of the main group were divided into two subgroups: with an NIHSS index value of < 10 (n = 10) and > 10 (n = 9). The control group included healthy donors with no history of acute and chronic inflammatory diseases (n = 8). Peripheral blood leukocytes were used as the test material. To determine expression of the TLR2 and TLR4 genes, RT-PCR in real time was used. Surface expression of TLRs was determined by flow cytometry. A study of the TLR2 and TLR4 gene expression showed that on the 1st, 3rd and 7th day post-stroke, the TLR4 gene expression in patients was significantly increased, when compared to the control group (p < 0.01), whereas TLR2 gene expression on the 3rd day of the disease was not statistically different from the control group. A study of surface expression of receptors showed that the average TLR2 fluorescence intensity on the patients’ peripheral blood monocytes was significantly increased on the 1st and 3rd day of disease when compared to the control group. The surface expression of TLR4 on monocytes has a statistically significant increase only on day 7. Assessment of surface expression of TLRs in subgroups with different severity values by NIHSS showed that patients with a NIHSS index > 10 had a significantly higher level of surface of TLR2 expression over the observation period, while the largest difference in TLR4 expression in the subgroups was observed on the 1st day of the disease (p < 0.05). Patients with ischemic stroke showed an increase in TLR2 and TLR4 expression at the gene and protein level, compared to healthy donors. These indices can be considered possible predictors for clinical prognosis of ischemic stroke.

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Inducible cAMP early repressor (ICER)-evoked delayed neuronal death in the organotypic hippocampal culture

Journal of Neuroscience Research ◽

10.1002/jnr.21469 ◽

2007 ◽

Vol 86 (1) ◽

pp. 61-70 ◽

Cited By ~ 9

Author(s):

Barbara Mioduszewska ◽

Jacek Jaworski ◽

Arek W. Szklarczyk ◽

Agata Klejman ◽

Leszek Kaczmarek

Keyword(s):

Neuronal Death ◽

Delayed Neuronal Death ◽

Hippocampal Culture ◽

Inducible Camp Early Repressor ◽

Organotypic Hippocampal Culture

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Abstract TP215: Machine Learning Analysis of Gene Expression Data and Predictive Stroke Outcomes in Acute Ischemic Stroke

Stroke ◽

10.1161/str.51.suppl_1.tp215 ◽

2020 ◽

Vol 51 (Suppl_1) ◽

Author(s):

Sarah R Martha ◽

Qiang Cheng ◽

Liyu Gong ◽

Lisa Collier ◽

Stephanie Davis ◽

...

Keyword(s):

Gene Expression ◽

Machine Learning ◽

Ischemic Stroke ◽

Acute Ischemic Stroke ◽

Mechanical Thrombectomy ◽

Stroke Patients ◽

Stroke Outcomes ◽

Inflammatory Genes ◽

Patient Demographics ◽

Edema Volume

Background and Purpose: The ability to predict ischemic stroke outcomes in the first day of admission could be vital for patient counseling, rehabilitation, and care planning. The Blood and Clot Thrombectomy Registry and Collaboration (BACTRAC; clinicaltrials.gov NCT03153683) collects blood samples distal and proximal to the intracranial thrombus during mechanical thrombectomy. These samples are a novel resource in evaluating acute gene expression changes at the time of ischemic stroke. The purpose of this study was to identify inflammatory genes and patient demographics that are predictive of stroke outcomes (infarct and/or edema volume) in acute ischemic stroke patients. Methods: The BACTRAC study is a non-probability, convenience sampling of subjects (≥ 18 year olds) treated with mechanical thrombectomy for emergent large vessel occlusion. We evaluated relative concentrations of mRNA for gene expression in 84 inflammatory molecules in static blood distal and proximal to the intracranial thrombus from adults who underwent thrombectomy. We employed a machine learning method, Random Forest, utilizing the first set of enrolled subjects, to predict which inflammatory genes and patient demographics were important features for infarct and edema volumes. Results: We analyzed the first 28 subjects (age = 66 ± 15.48, 11 males) in the BACTRAC registry. Results from machine learning analyses demonstrate that the genes CCR4, IFNA2, IL9, CXCL3, Age, DM, IL7, CCL4, BMI, IL5, CCR3, TNF, and IL27 predict infarct volume. The genes IFNA2, IL5, CCL11, IL17C, CCR4, IL9, IL7, CCR3, IL27, DM, and CSF2 predict edema volume. There is an intersection of genes CCR4, IFNA2, IL9, IL7, IL5, CCR3 to both infarct and edema volumes. Overall, these genes depicts a microenvironment for chemoattraction and proliferation of autoimmune cells, particularly Th2 cells and neutrophils. Conclusions: Machine learning algorithms can be employed to develop predictive biomarker signatures for stroke outcomes in ischemic stroke patients, particularly in regard to identifying acute gene expression changes that occur during stroke.

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