Genetic risk impacts stroke mortality and pathogenesis in patients with ischemic stroke: a cohort study of BioBank Japan

Don't you dare use genetic risk as an excuse for not changing the status quo and getting these people recovered! Excuses won't be tolerated, you're fired! Leaders would solve such a problem; obviously you have NO LEADERSHIP potential!

Genetic risk impacts stroke mortality and pathogenesis in patients with ischemic stroke: a cohort study of BioBank Japan

Takashi Shimoyama^1*†Yoichiro Kamatani^2†Koichi Matsuda^3†Hiroki Yamaguchi^4†Kazumi Kimura^1†

• ¹Department of Neurology, Nippon Medical School, Tokyo, Japan
• ²Laboratory of Complex Trait Genomics, Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan
• ³Department of Computational Biology and Medical Science, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan
• ⁴Department of Hematology, Nippon Medical School, Tokyo, Japan

Background: Previous multi-ancestry genome-wide association studies (GWAS) of stroke reported 32 stroke risk loci in the MEGASTROKE study. Most studies on the genetic risk score (GRS) of stroke have reported a predominance in the European general population. We aimed to explore the association among GRS, clinical characteristics, and mortality in patients with ischemic stroke registered in the BioBank Japan (BBJ) database.

Methods: This is a cohort study of BBJ participants. The project participants were recruited between June 2003 and March 2018. We conducted a GWAS for stroke in 19,702 Japanese patients with ischemic stroke and 159,610 controls. GRS was generated using 29 stroke risk single nucleotide polymorphisms (SNPs) from 32 stroke-related loci identified in the MEGASTROKE. A multivariate logistic regression model was used to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs) for comorbidities and stroke etiology across the GRS. The Cox proportional hazard model was used to estimate hazard ratios (HRs) and 95% CIs for mortality associated with GRS.

Results: The ORs for atrial fibrillation were significantly higher in those at Intermediate GRS [20–80th percentile of GRS; ORs 1.59 (1.25–1.90)] and High GRS [top 20th percentile of GRS; ORs 2.12 (1.69–2.67)] after a full adjustment than in those at Low GRS (bottom 20th percentile of GRS). Regarding stroke etiology, the ORs for cardioembolism were significantly higher in those at Intermediate GRS [ORs 1.31 (1.04–1.61)] and High GRS [ORs 1.44 (1.13–1.89)] than in those at Low GRS. During a median follow-up of 10.0 years, the risk of stroke mortality was significantly higher in those at High GRS [HRs 1.27 (1.04–1.56)] than in those at Low GRS in a fully adjusted model.

Conclusion: In Japanese, a higher GRS was significantly associated with atrial fibrillation, cardioembolism, and stroke mortality. Our findings suggest that the GRS may predict the risk of stroke mortality and provide insights into the pathogenesis of stroke.

Introduction

Stroke is the second-leading cause of death and the primary cause of neurological disability worldwide (1, 2). Stroke is caused by a complex interplay of environmental and traditional risk factors, including older age, hypertension, diabetes mellitus, dyslipidemia, atrial fibrillation, chronic kidney disease, and smoking (3). Besides conventional clinical risk factors, the genetic contribution to the development of stroke is also widely recognized (4). Twin and family history studies suggest genetic factors are responsible for some of this unexplained risk for stroke. The heritability estimates were 0.32 for the liability to stroke death and 0.17 for stroke hospitalization or stroke death (5).

Over the last decades, several genome-wide association studies (GWAS) have identified genetic variants associated with stroke in different ethnic populations (6–11). Previous multi-ancestry GWAS of 52,000 subjects in predominantly European-ancestry groups have identified 32 loci associated with stroke and stroke subtypes (MEGASTROKE study) (11). Recent work has highlighted the potential of the genetic risk score (GRS) based on the MEGASTROKE study, which can be evaluated as a risk factor for stroke and used to predict incident stroke events in an independent population (11–16). The risk of incident stroke was higher in those at high genetic risk than in those at low genetic risk (12). The polygenic risk score (PRS) using 3.6 million genetic variants predicts stroke incidents in a population of 12,792 healthy older individuals enrolled in the ASPREE trial (Aspirin in Reducing Events in the Elderly) (13). In a genetic cohort analysis pooling 51,288 subjects with cardiometabolic disease from five cardiovascular clinical trials, GRS using the set of 32 single nucleotide polymorphisms (SNPs) derived from the MEGASTROKE study was a strong, independent predictor of ischemic stroke incidence over a median follow-up period of 2.5 years (14). In the Northern Finland Birth Cohort 1966 of 12,058 children, higher PRS for stroke was associated with the risk for cerebrovascular disease in mid-life in Finnish population. Ischemic stroke (15). Although investigation of genetic risk for stroke has been limited in non-European populations, the Hisayama Study, which involved 3,038 Japanese individuals, reported the PRS for stroke using 350,000 SNPs was significantly associated with stroke incidence during long-term follow-up (median 10.2 years) (16). Most of the advanced literature on genetic risk for stroke has been reported in general populations regardless of ethnicity; however, solid evidence in the relevant literature has not described the clinical significance of genetic risk for stroke in non-European stroke patients.

To address these limitations, we developed a GRS for stroke from a set of 32 stroke risk loci identified in the MEGASTROKE study in Japanese patients with ischemic stroke. We hypothesized that subsets with a higher genetic risk influence stroke mechanisms and mortality compared to those with a lower genetic risk of ischemic stroke. This cohort study aimed to clarify the association between the GRS score, clinical characteristics, and mortality in stroke patients registered in the BioBank Japan (BBJ) database.

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Analysis of the expression level and predictive value of CLEC16A|miR-654-5p|RARA regulatory axis in the peripheral blood of patients with ischemic stroke based on biosignature analysis

 Why the fuck are you doing predictions rather than the work that will get survivors recovered?

Laziness? Incompetence? Or just don't care? NO leadership? NO strategy? Not my job? Not my Problem?

Analysis of the expression level and predictive value of CLEC16A|miR-654-5p|RARA regulatory axis in the peripheral blood of patients with ischemic stroke based on biosignature analysis

Jiang-jie Hao¹Yuan Liu¹Jun-hua Lu²Ying Zhao³Ying Lin³Li-qiu Ma³Ping Xue³Bao-yun Jin¹Bei-bei Li¹Zheng Zhou¹Xin-xin Huang¹Hong-jun Guan^1*

• ¹Department of Public Health, Mudanjiang Medical University, Mudanjiang, China
• ²Department of Nursing, Mudanjiang Medical University, Mudanjiang, China
• ³Hongqi Hospital Affiliated to Mudanjiang Medical University, Mudanjiang, China

Introduction: Ischemic stroke (IS) is a cerebrovascular disease that can be disabling and fatal, and there are limitations in the clinical treatment and prognosis of IS. It has been reported that changes in the expression profile of circRNAs have been found during injury in ischemic stroke, and circRNAs play an important role in the IS cascade response. However, the specific mechanisms involved in the pathogenesis of IS are not yet fully understood, and thus in-depth studies are needed.

Methods: In this study, one circRNA dataset (GSE161913), one miRNA dataset (GSE60319) and one mRNA dataset (GSE180470) were retrieved from the Gene Expression Omnibus (GEO) database and included, and the datasets were differentially expressed analyzed by GEO2R and easyGEO to get the DEcircRNA, DEmiRNA and DEmRNA, and DEmRNA was enriched using ImageGP, binding sites were predicted in the ENCORI database, respectively, and the competitive endogenous RNA (ceRNA) regulatory network was visualized by the cytoscape software, and then selected by MCC scoring in the cytoHubba plugin Hub genes. In addition, this study conducted a case–control study in which blood samples were collected from stroke patients and healthy medical examiners to validate the core network of ceRNAs constructed by biosignature analysis by real-time fluorescence quantitative qRT-PCR experiments.

Results: A total of 233 DEcircRNAs, 132 DEmiRNAs and 72 DEmRNAs were screened by bioinformatics analysis. circRNA-mediated ceRNA regulatory network was constructed, including 148 circRNAs, 43 miRNAs and 44 mRNAs. Finally, CLEC16A|miR-654-5p|RARA competitive endogenous regulatory axis was selected for validation by qRT-PCR, and the validation results were consistent with the bioinformatics analysis.

Discussion: In conclusion, the present study establishes a new axis of regulation associated with IS, providing new insights into the pathogenesis of IS.

1 Introduction

Stroke is a type of cerebrovascular disease in which a cerebral blood vessel suddenly ruptures or becomes blocked, resulting in insufficient blood supply to the brain and thus causing severe brain tissue damage and loss of neuronal function. It is one of the leading causes of mortality and morbidity worldwide and is a chronic non-communicable disease that poses a serious threat to the health of the global population. Ischemic stroke (IS) is one of the two main categories of stroke (1), commonly known as cerebral infarction, and refers to the occlusion of the arteries supplying the brain, which leads to necrosis of brain tissue and focal neurological deficits. Ischemic strokes occur every year, and most of them are in the elderly, and can be seriously disabling and fatal (2), causing a heavy medical burden on patients, families and society. According to the data published by the World Health Organization (WHO), the incidence of stroke is getting younger globally, which is a disturbing trend as people’s knowledge about stroke is increasing. Therefore, the prevention and treatment of ischemic stroke should be given sufficient attention to elucidate its pathogenesis as soon as possible and develop solutions to improve the quality of life of the population.

The current diagnosis of stroke relies on clinical symptoms and medical imaging techniques. The most commonly used diagnostic tools for IS are computed tomography (CT) and magnetic resonance (MR) (3). However, CT has poor sensitivity to early ischemic changes in the brain and most patients do not show typical imaging changes in the early stages of the disease, which may miss the optimal time for treatment. Restoration of cerebral blood flow supply is currently the main therapeutic strategy for acute cerebrovascular disease, and early re-establishment of cerebral blood supply by revascularization techniques of intravenous thrombolysis or arterial thrombolysis is the most effective clinical treatment for IS (4). However, there are limitations in the duration of treatment and the application of techniques, and most patients are left with irreversible brain tissue damage with the risk of hemorrhage and ischemia–reperfusion injury (5). Therefore, it is urgent to explore a rapid and accurate biomarker for early diagnosis and prediction of IS, and it is important to further elucidate the pathophysiological mechanisms of ischemic stroke and find timely and effective therapeutic targets.

In recent years, many studies have reported that changes in the expression profiles of noncoding RNAs (ncRNAs) were found during the injury process in ischemic stroke. miRNAs are a class of ubiquitous single-stranded non-coding RNAs that can specifically bind to messenger RNAs (mRNAs) through base complementary pairing, interfering with the translational process and thus inhibiting or altering protein synthesis. Numerous studies have shown that miRNAs, as important mediators of gene regulation, play important roles in IS cascade reactions. Translateral ventricular injection of miR-377 inhibitor in MCAO rats attenuates ischemic brain injury by promoting angiogenesis and inhibiting brain inflammation induced by pro-inflammatory factor release (6). miR-455-5p expression is downregulated in brain tissue and peripheral blood after cerebral ischemia/reperfusion (I/R), and overexpression of miR-455-5p reduces neuroinflammation through inhibition of CCR5 expression, thereby attenuating I/R-induced injury (7). Endothelial miR-15a/16–1 is a negative regulator of cerebral angiogenesis and neurological recovery after stroke (8).

Circular RNA (circRNA) is a unique structure produced by reverse splicing with a high degree of stability, interspecies conservation, and tissue expression specificity, making circRNA an ideal molecular biomarker for diagnosis (9). With the development of bioinformatics analysis methods, many scholars have identified differences in the expression of circRNAs in different organisms and tissues. In a study to identify differentially expressed circRNAs in blood samples from Acute IS patients and healthy controls by circRNA microarray, three circRNAs (circFUNDC1, circPDS5B, and circCDC14A) were up-regulated in AIS patients compared to healthy subjects, and the ROC curve showed an area under the curve (AUC) of 0.875, with a specificity of 91% and a sensitivity of 71.5%, indicating that these 3 circRNAs have diagnostic and prognostic value for AIS (10). The expression of circFOXP1 was significantly reduced in the peripheral blood of AIS patients, and overexpression of circFOXP1 could attenuate post-ischemic brain injury by regulating the STAT3/apoptosis signaling pathway (11). Studies have shown that circRNAs can act as miRNA sponges to regulate gene expression, interact with proteins to perform important biological functions, and encode proteins as translational templates. In recent years, the competitive endogenous RNA (ceRNA) mechanism of circRNAs has been extensively studied. Briefly, circRNAs chelate miRNAs through spongy competition and regulate the inhibitory effect of miRNAs on base complementary pairing at target sites in the untranslated region of messenger RNAs (mRNAs), which in turn regulates the expression of downstream target genes (12). A number of studies have identified a close association between circRNA-mediated ceRNA regulatory networks and ischemic stroke pathophysiological processes (13). A study demonstrated that the expression of circ-HECTD1 and tumor necrosis factor receptor-associated factor 3 (TRAF3) was significantly upregulated in ischemic brain tissues, whereas the expression of miR-133b was downregulated. Knockdown of circ-HECTD1 attenuated neuronal damage induced by cerebral ischemia by targeting binding to miR-133b and inhibiting TRAF3 expression, thereby inhibiting OGD-induced apoptosis and NF-κB activation (14). SNHG15 is upregulated in hypoxic–ischemic mice or cellular models, and inhibition of SNHG15 expression ameliorates ischemia-hypoxia-induced neuronal injury and microglial cell inflammation via the miR-302a-3p / STAT1 / NF-κB pathway (13). Han et al. (15) showed that CircHECTD1 levels were significantly increased in a transient middle cerebral artery occlusion (TMCAO) mouse stroke model, which was validated in plasma samples from AIS patients. By interfering with CircHECTD1 expression using an siRNA approach, MIR142 was released and accompanied by the downstream down-regulation of TIPARP expression, which resulted in the improvement of cerebral infarction by inhibiting astrocyte activation. In summary, circRNA is closely related to the pathophysiological process of ischemic stroke. As the abnormally expressed circRNA molecules are continuously mined for neuroprotective or deleterious effects on IS through the ceRNA regulatory pathway, the specific mechanisms of circRNA and stroke onset, progression and prognosis will be gradually clarified.

In this paper, we take ischemic stroke as an entry point to reveal the pathogenesis of ischemic cerebrovascular diseases from the perspective of genomics, so as to provide more valuable reference information for the clinic. In this study, we obtained circRNA, miRNA and mRNA expression datasets from the Gene Expression Omnibus (GEO) database, screened the differentially expressed genes and constructed the IS-associated ceRNA network, and then extracted a regulatory axis from them by combining with the Protein–protein interaction (PPI) network analysis, and then verified the gene expression levels through reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and the results of the present study may provide a new idea for elucidating the potential mechanisms underlying the occurrence and development of ischemic stroke. The flow chart illustrating the steps of the whole analysis was shown in Figure 1.

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OPIOID ANTAGONISTS AS POTENTIAL THERAPEUTICS FOR ISCHEMIC STROKE

For discussion with your doctor.

Opioid antagonists are a class of compounds that selectively bind to opioid receptors. These compounds inhibit opioid receptor activation by blocking the actions...

OPIOID ANTAGONISTS AS POTENTIAL THERAPEUTICS FOR ISCHEMIC STROKE

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Highlights

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Prescription opioids exacerbate risk and severity of ischemic stroke.

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This perspective addresses a novel application and repurposing of FDA-approved opioid antagonists to minimize BBB damage, reduce stroke severity, and promote neural recovery.

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Future directions discuss potential drug design and delivery methods to enhance these novel therapeutic targets.

Abstract

Chronic use of prescription opioids can exacerbate risk and severity of ischemic stroke. Annually, 6 million people die from stroke worldwide and there are no neuroprotective or neurorestorative agents to improve stroke outcomes and promote recovery. Prescribed opioids such as morphine have been shown to alter tight junction protein expression, resulting in the disruption of the blood brain barrier (BBB), ultimately leading to stroke pathogenesis. Consequently, protection of the of BBB has been proposed as a therapeutic strategy for ischemic stroke. This perspective addresses the deficiency in stroke pharmacological options and examines a novel application and repurposing of FDA-approved opioid antagonists as a prospective neuroprotective therapeutic strategy to minimize BBB damage, reduce stroke severity, and promote neural recovery. Future directions discuss potential drug design and delivery methods to enhance these novel therapeutic targets.

Graphical abstract

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