# Identification of Differentially Expressed microRNAs Associated with Ischemic Stroke by Integrated Bioinformatics Approaches

Authors: Shengqiang Jiang; Jie Wu; Yan Geng; Yuting Zhang; Yupeng Wang; Jinrong Wu; Chunqu Lu; Guoxuan Luo; Jie Zan; Yong Zhang
Journal: International Journal of Genomics
 (2022)

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## Abstract

Ischemic stroke (IS) is one of the leading causes of disability and mortality worldwide. This study aims to find the crucial exosomal miRNAs associated with IS by using bioinformatics methods, reveal potential biomarkers for IS, and investigate the association between the identified biomarker and immune cell pattern in the peripheral blood of IS patients. In this study, 3 up-regulated miRNAs (hsa-miR-15b-5p, hsa-miR-184, and hsa-miR-16-5p) miRNAs in the serum exosomes between IS patients and healthy controls from GEO database (GSE199942) and 25 down-regulated genes of peripheral blood mononuclear cells of IS patients from GSE22255 were obtained with the help of the R software. GO annotation and KEGG pathway enrichment analysis showed that the 25 down-regulated genes were associated with coenzyme metabolic process and were mainly enriched in the N-glycan biosynthesis pathway. Furthermore, we performed the LASSO algorithm to narrow down the above 25 intersected genes, and identified 8 key genes which had a good diagnostic value in discriminating IS patients from the healthy controls analyzed with ROC curve. CIBERSORT algorithm indicated that the abundance of M0 macrophages and resting mast cells was significantly lower than that of the control group. The spearman correlation analysis showed that STT3A was negatively correlated with the proportion of follicular helper T cells, activated NK cells and resting dendritic cells. Finally, GSE117064 showed that has-miR-16-5p was more advantageous for diagnosing stroke. In conclusion, hsa-miR-15b-5p, hsa-miR-184, and hsa-miR-16-5p are identified as specific related exosomal miRNAs for IS patients. These genes may provide new targets for the early identification of IS.


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## Body

## 1. Introduction

Ischemic stroke (IS) is one of the leading causes of disability and mortality worldwide and accounts for 80% of all strokes [1]. Intravenous thrombolysis is the only approved treatment strategy for acute IS by FDA [2]. Due to the short treatment time window within 4.5 h of onset and the high risk of hemorrhage, the thrombolysis rate in IS patients is very low [3], suggesting early diagnosis is extremely critical. Nowadays, clinical diagnosis of IS mainly relies on magnetic resonance imaging and computed tomography [4]. However, most community medical institutions lack the testing equipment, and the brain imaging examinations are relatively expensive, which limits the clinical diagnosis of IS [5]. Thus, developing new diagnostic markers and therapeutic targets for IS are urgently needed.microRNAs (miRNAs) are a class of small and noncoding RNA, which regulate gene expression through silencing target genes and play important roles in the nervous system diseases, such as traumatic brain injury, spinal cord injury, subarachnoid hemorrhage, and IS [6–8]. For example, miR-31 inhibits traumatic brain injury-triggered neuronal cell apoptosis by regulating hypoxia-inducible factor-1A/vascular endothelial growth factor A axis [7]. miR-672-3p promotes functional recovery in rats with contusive spinal cord injury by inhibiting ferroptosis suppressor protein 1 [8]. Nowadays, more and more studies identify that some aberrant expressed circulating miRNAs in the serum, which are identified via bioinformatics methods, could be used as potential candidates for disease diagnosis. For example, Hsa-miR-484, hsa-miR-185-5p, hsa-miR-340-5p, hsa-miR-146a-5p, and hsa-miR-195-5p have a prognostic value in breast cancer patients treated with integrative interventions, including diet and physical activity [9]. Specific alteration of 20 miRNAs has a direct association with pesticide exposure and the development of neurodegenerative diseases [10]. Hsa-miR-181a-3p, hsa-miR-214-3p, hsa-miR-18a-5p, and hsa-miR-938 are positively related to the IL-6 signaling pathway activation in various cancers [11]. 14 potentially important miRNAs are identified as IS diagnostic signatures via bioinformatics method combined with logistic regression analysis [12]. Whereas circulating miRNAs in the serum are generally affected by the different pathophysiological conditions and circulating ribonucleases [13], there are still short of reliable prognostic or diagnostic blood biomarkers.Exosomes are spherical extracellular nanovesicles with a bilayer lipid structure for the outer membrane, defined by a diameter of 30–100 nm [14]. Exosomes contain proteins, mRNAs, and miRNAs and play essential roles in intercellular communication [15]. Due to their high stability, serum exosomal miRNAs are considered to be powerful non-invasive biomarkers in many diseases, including breast cancer, pancreatic ductal adenocarcinoma, and hepatocellular carcinoma [16–18]. Recently, Tong et al. [19] found that miR-151a-5p, miR-24, mir-485-5p, mir-331-5p, and mir-214 were upregulated with statistical significance in both the serum exosome and cerebrospinal fluid exosomes, and may be considered as a biomarker for the diagnosis of Parkinson’s disease.In this study, we attempt to identify the crucial exosomal miRNAs associated with IS by using bioinformatics methods, reveal potential biomarkers for IS, and investigate the association between the identified biomarker and immune cell infiltration in IS.


## 2. Materials and Methods

### 2.1. Data Sources

The data were downloaded from the Gene Expression Omnibus (GEO) database (http://www.ncbi.nlm.nih.gov/geo). The inclusion and exclusion criteria for these data sets were as follows. Inclusion criteria: (i) data sets containing miRNA and mRNA expression levels of IS patients; (ii) data sets reporting miRNAs expression levels of both IS patient and healthy patient; and (iii) data sets containing miRNA expression data of at least 5 IS samples and 5 controls. Exclusion criteria: (i) data sets containing exclusively IS sample and (ii) data sets containing miRNAs expression levels of animal models, cell lines or other in vitro experiments.GSE199942 included the miRNA expression profiles of serum exosomes in five acute IS patients and five healthy controls detected by RNA-seq using llumina HiSeqTM 2500. GSE22255 included the gene expression profiling in peripheral blood mononuclear cells (PBMCs) of 20 IS patients and 20 sex- and age-matched controls using Affymetrix microarrays. GSE16561 included the gene expression profiles of peripheral whole blood RNA from 39 IS patients and 24 healthy controls. GSE117064 included the microRNA profiles of 1785 samples, which consist of 173 of CVD patients, 1612 of non-CVD control using microarrays. PMC8702168 included the miRNA expression profiles of human stroke brain tissue in five acute IS patients and three non-stroke controls detected by RNA-seq using Illumina NextSeq 500. All the data sets used in this study could be available on GEO data sets.


### 2.2. Analysis of Differentially Expressed miRNAs and Genes

The differentially expressed miRNAs from GSE199942 were obtained using the edgeR package [20]. The differentially expressed genes from GSE22255 were obtained using the limma package [21]. The P value <0.05 was used as threshold for nominally significant differential expression. gplot2 and pheatmap packages were used to draw the volcanic maps and heat maps, respectively.


### 2.3. Sankey Diagram of the miRNAs-mRNAs Network

The miRNAs-mRNAs network was drawn using the “ggalluvial” package of R software.


### 2.4. GO and KEGG Analysis

Gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were performed using the “clusterProfiler” R package [22]. P value <0.05 was considered the criterion for statistical significance.


### 2.5. Evaluation of Immune Cell Abundance

The gene expression profiling of GSE22255 was used to compare immune cell abundance in the peripheral blood of 20 IS patients and 20 healthy controls. The “CIBERSORT” algorithm was used to calculate the relative proportions of 22 types of immune cells [23]. Significant alterations in immune cells were identified using the Wilcoxon test at P<0.05. The “ggcorrplot” package was used to visualize the results of correlation between immune cells.


### 2.6. Diagnostic Value of Differentially Expressed Genes in IS

The receiver operating characteristic (ROC) curve analysis is often used to determine the best diagnostic threshold for a diagnostic method. Generally, the area under the curve (AUC) value of >0.7 obtained from ROC curve analysis is considered to have a good predictive value [24].


## 2.1. Data Sources

The data were downloaded from the Gene Expression Omnibus (GEO) database (http://www.ncbi.nlm.nih.gov/geo). The inclusion and exclusion criteria for these data sets were as follows. Inclusion criteria: (i) data sets containing miRNA and mRNA expression levels of IS patients; (ii) data sets reporting miRNAs expression levels of both IS patient and healthy patient; and (iii) data sets containing miRNA expression data of at least 5 IS samples and 5 controls. Exclusion criteria: (i) data sets containing exclusively IS sample and (ii) data sets containing miRNAs expression levels of animal models, cell lines or other in vitro experiments.GSE199942 included the miRNA expression profiles of serum exosomes in five acute IS patients and five healthy controls detected by RNA-seq using llumina HiSeqTM 2500. GSE22255 included the gene expression profiling in peripheral blood mononuclear cells (PBMCs) of 20 IS patients and 20 sex- and age-matched controls using Affymetrix microarrays. GSE16561 included the gene expression profiles of peripheral whole blood RNA from 39 IS patients and 24 healthy controls. GSE117064 included the microRNA profiles of 1785 samples, which consist of 173 of CVD patients, 1612 of non-CVD control using microarrays. PMC8702168 included the miRNA expression profiles of human stroke brain tissue in five acute IS patients and three non-stroke controls detected by RNA-seq using Illumina NextSeq 500. All the data sets used in this study could be available on GEO data sets.


## 2.2. Analysis of Differentially Expressed miRNAs and Genes

The differentially expressed miRNAs from GSE199942 were obtained using the edgeR package [20]. The differentially expressed genes from GSE22255 were obtained using the limma package [21]. The P value <0.05 was used as threshold for nominally significant differential expression. gplot2 and pheatmap packages were used to draw the volcanic maps and heat maps, respectively.


## 2.3. Sankey Diagram of the miRNAs-mRNAs Network

The miRNAs-mRNAs network was drawn using the “ggalluvial” package of R software.


## 2.4. GO and KEGG Analysis

Gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were performed using the “clusterProfiler” R package [22]. P value <0.05 was considered the criterion for statistical significance.


## 2.5. Evaluation of Immune Cell Abundance

The gene expression profiling of GSE22255 was used to compare immune cell abundance in the peripheral blood of 20 IS patients and 20 healthy controls. The “CIBERSORT” algorithm was used to calculate the relative proportions of 22 types of immune cells [23]. Significant alterations in immune cells were identified using the Wilcoxon test at P<0.05. The “ggcorrplot” package was used to visualize the results of correlation between immune cells.


## 2.6. Diagnostic Value of Differentially Expressed Genes in IS

The receiver operating characteristic (ROC) curve analysis is often used to determine the best diagnostic threshold for a diagnostic method. Generally, the area under the curve (AUC) value of >0.7 obtained from ROC curve analysis is considered to have a good predictive value [24].


## 3. Results

### 3.1. Identification of Differentially Expressed Exosomal miRNAs Related to IS

In this study, we analyzed the differentially expressed miRNAs in serumexosomes between IS patients and healthy controls from GSE199942 data sets, and found that 35 upregulated and 24 downregulated miRNAs (Table 1). The expression of these miRNAs is shown in Figures 1(a) and 1(b). Furthermore, the differentially expressed serum exosomes miRNAs were intersected with those differentially expressed miRNAs from freshly removed human stroke brain tissue [25], and three upregulated miRNAs were obtained: hsa-miR-15b-5p, hsa-miR-184, and hsa-miR-16-5p (Figures 1(c) and 1(d)).Table 1
The differentially expressed miRNAs in serum exosomes between IS patients and healthy controls from GSE199942 data sets.
GeneslogFClogCPMP ValueFDRChangehsa-miR-45114.01777513.15193110.0003910.1276672Uphsa-miR-450a-5p2.36085314.750.00187180.2345272Uphsa-miR-15b-5p1.87015655.68244970.00275740.2345272Uphsa-miR-187-3p6.04097993.63702480.00287320.2345272Uphsa-miR-44883.1070225.14011730.00490580.2367778Uphsa-miR-30a-5p2.460329210.2023890.00518250.2367778Uphsa-miR-3688-3p4.51277811.92195090.00548930.2367778Uphsa-miR-30a-3p2.34924787.31374970.00571830.2367778Uphsa-miR-519b-5p5.55156392.28563430.00677650.2367778Uphsa-miR-523-5p5.55156352.28563430.00677690.2367778Uphsa-miR-522-5p5.55152362.28563430.00681490.2367778Uphsa-miR-518e-5p5.551472.28563430.00686640.2367778Uphsa-miR-519c-5p5.55145372.28563430.00688220.2367778Uphsa-miR-519a-5p5.55138482.28563430.00694910.2367778Uphsa-miR-23a-3p1.39001298.91372210.00731520.2367778Uphsa-miR-223-3p1.69909386.59243470.00761460.2367778Uphsa-miR-12624.76793631.84184620.00903330.2564662Uphsa-miR-1843.92838663.48733810.00964320.2623126Uphsa-let-7g-3p4.7771191.83496660.0108460.2623126Uphsa-miR-503-5p1.46372555.97198430.0116940.2727198Uphsa-miR-2115-3p3.35989632.57556350.01412690.3180986Uphsa-miR-365a-3p3.37513443.19586650.02101960.4111132Uphsa-miR-365b-3p3.37521763.19586650.02133120.4111132Uphsa-miR-143-3p1.310620212.3402440.02321670.4123441Uphsa-miR-140-3p1.100650413.623170.02399550.4123441Uphsa-miR-4677-3p2.65116542.77538440.02470740.4136901Uphsa-miR-708-3p1.58378764.8998950.02688260.4341984Uphsa-miR-518d-5p4.59331271.77068090.02844910.4341984Uphsa-miR-520c-5p4.59329671.77068090.02848860.4341984Uphsa-miR-526a4.59325491.77068090.02859190.4341984Uphsa-miR-16-5p1.181193710.0140530.02954330.4384491Uphsa-miR-873-5p2.73028293.65443560.03059880.4440228Uphsa-miR-218-5p1.6730554.87193180.03272280.4546377Uphsa-miR-542-3p1.7027244.00296470.03874680.4914672Uphsa-miR-501-5p2.64086561.93044820.04660.5261696Uphsa-miR-185-5p–0.99593414.1610130.05117420.5569459No significant changehsa-miR-206–1.3927415.74629870.06334470.6573682No significant changehsa-miR-7113-5p3.80997031.66120460.06525890.6573682No significant changehsa-miR-509-3-5p3.34645992.10642680.06703690.6573682No significant changehsa-miR-548ad-5p1.59152794.23046390.06794660.6573682No significant changehsa-miR-548ae-5p1.59152354.23046390.06844970.6573682No significant changehsa-miR-1070.96472057.85669260.0698410.6573682No significant changehsa-miR-6511b-5p–1.9091833.26213640.07168490.6573682No significant changehsa-miR-6741-3p–2.2649892.29295630.07231310.6573682No significant changehsa-miR-145-3p1.03984946.72405290.07234870.6573682No significant changehsa-miR-204-3p1.64785425.2089780.07284230.6573682No significant changehsa-miR-330-3p–0.8693867.17905010.07312150.6573682No significant changehsa-miR-4762-3p–2.7991182.00769360.07543120.6573682No significant changehsa-miR-589-5p0.86365026.14292560.07737010.6573682No significant changehsa-miR-671-3p–1.0410895.14095410.07906740.6573682No significant changehsa-miR-323b-3p2.22656373.02135460.07929160.6573682No significant changehsa-miR-125a-5p–1.1123567.76783990.07983320.6573682No significant changehsa-miR-141-3p–2.5778722.36672620.08009670.6573682No significant changehsa-miR-3913-5p1.09226744.84461920.08138340.6573682No significant changehsa-miR-378c–1.3333669.24488390.08148840.6573682No significant changehsa-miR-744-5p–0.7978289.55633710.08229310.6573682No significant changehsa-miR-148a-5p–1.1023755.65354380.08422960.6573682No significant changehsa-miR-5193–2.9965551.73410560.0843130.6573682No significant changehsa-miR-1224-5p–1.3338085.02872360.08568650.6573682No significant changehsa-miR-1298-5p2.12718152.45418490.08632590.6573682No significant changehsa-miR-548ay-5p1.4805353.46988450.08859520.6573682No significant changehsa-miR-326–1.4902043.24920.08886160.6573682No significant changehsa-miR-4755-5p–1.9634282.98187390.08897110.6573682No significant changehsa-miR-331-5p1.36015363.33888490.08912020.6573682No significant changehsa-miR-27a-5p0.91835896.58580810.09147640.6573682No significant changehsa-miR-3928-3p–1.2718815.10733890.09309950.6573682No significant changehsa-miR-509-3p3.10730882.47489860.09352760.6573682No significant changehsa-miR-3157-3p1.64744042.53547580.09392050.6573682No significant changehsa-miR-6810-5p–4.8127791.80994670.09414760.6573682No significant changehsa-miR-27a-3p1.01440349.78331950.09462880.6573682No significant changehsa-miR-15b-3p1.87011933.3805780.09688370.6590269No significant changehsa-miR-34a-5p4.16864581.60791170.09770920.6590269No significant changehsa-miR-3130-3p1.50385183.60046640.0994970.6590269No significant changehsa-miR-371a-5p–4.3373541.63136310.09955310.6590269No significant changehsa-miR-1292-5p–1.0331455.80122750.10072790.6590269No significant changehsa-miR-214-3p1.69976022.98037980.10177580.6590269No significant changehsa-miR-1307-3p–0.86251310.957750.10193220.6590269No significant changehsa-miR-12461.068577211.1201330.10603790.6612336No significant changehsa-miR-6747-3p–2.4033271.99047870.10871320.6612336No significant changehsa-miR-6857-3p–2.3131141.84542110.10937970.6612336No significant changehsa-miR-6859-5p–1.2934823.01404160.10991690.6612336No significant changehsa-miR-2277-3p–4.4861881.6844030.11049250.6612336No significant changehsa-miR-1273h-3p–1.0244394.77443060.11251730.6612336No significant changehsa-miR-4489–1.6526562.81154040.11572860.6612336No significant changehsa-miR-2110–0.7633999.5335660.11601440.6612336No significant changehsa-miR-582-3p0.96925916.87174130.11788040.6612336No significant changehsa-miR-4211.13338573.76034650.11796840.6612336No significant changehsa-miR-6808-3p–1.8642982.03410990.1182890.6612336No significant changehsa-miR-6855-5p2.32068252.69201970.11851410.6612336No significant changehsa-miR-30d-3p1.96583832.23801370.11857740.6612336No significant changehsa-miR-548g-3p2.13659631.92773460.12029410.6612336No significant changehsa-miR-4632-5p–4.4384521.66466510.12060230.6612336No significant changehsa-miR-223-5p1.032683810.9945970.12236150.6612336No significant changehsa-miR-148b-3p0.85718429.11758560.12309290.6612336No significant changehsa-miR-511-5p–0.9542877.69622270.12339120.6612336No significant changehsa-miR-3120-5p–1.5695493.1895440.12389890.6612336No significant changehsa-miR-6807-5p–1.684452.76563620.12580880.6612336No significant changehsa-miR-7706–0.8009768.13641830.12598730.6612336No significant changehsa-miR-6877-5p–0.978644.03466950.12817990.6612336No significant changehsa-miR-6867-5p–4.4203871.63993480.1287130.6612336No significant changehsa-miR-654-3p1.47164824.0584580.12927710.6612336No significant changehsa-miR-532-3p1.33452663.05794250.13109350.6612336No significant changehsa-miR-139-5p–0.7639618.79830530.13110840.6612336No significant changehsa-miR-150-5p–0.9517214.57762610.13150270.6612336No significant changehsa-miR-29c-3p1.5905682.93742160.13218380.6612336No significant changehsa-miR-216b-5p2.23739782.2036770.13259360.6612336No significant changehsa-miR-182-5p0.96614188.88975240.13337840.6612336No significant changehsa-miR-422a–1.0784674.62985080.13380460.6612336No significant changehsa-miR-598-5p–2.6666131.57353920.1346770.6612336No significant changehsa-miR-5001-5p–2.5515532.03696420.13704730.6620332No significant changehsa-miR-134-5p–0.9807169.42155160.13734060.6620332No significant changehsa-miR-12842.41454062.19685350.13788130.6620332No significant changehsa-miR-3150a-3p–2.8117731.56347350.13904330.6627395No significant changehsa-miR-363-3p0.70737629.10435110.14135060.6688546No significant changehsa-miR-1228-5p–1.5837185.62672190.15101840.6974406No significant changehsa-miR-451b1.55565342.3416550.15134680.6974406No significant changehsa-miR-4690-3p–1.5853322.31226930.15460860.6974406No significant changehsa-miR-44921.88907492.69085750.15561860.6974406No significant changehsa-miR-22-5p0.71134847.50086180.15644730.6974406No significant changehsa-miR-382-3p2.66187061.88842540.15693760.6974406No significant changehsa-miR-1268a–1.3440743.51730160.1572150.6974406No significant changehsa-miR-130b-5p–1.3498465.53683020.15731530.6974406No significant changehsa-miR-1908-5p–0.8683765.32057330.15750610.6974406No significant changehsa-miR-13233.17542721.74996440.15871520.6974406No significant changehsa-miR-142-5p0.924594310.7469010.15914030.6974406No significant changehsa-miR-202-5p–2.1482192.10916040.16118120.7016757No significant changehsa-miR-3614-5p0.82358815.102390.16282950.7029907No significant changehsa-miR-4734–2.3886632.05939320.16363640.7029907No significant changehsa-miR-500a-3p0.81909936.92276090.16712410.7088167No significant changehsa-miR-6837-3p2.33387041.53371010.16716350.7088167No significant changehsa-miR-1251-5p3.91765011.53416430.17145350.7155546No significant changehsa-miR-1303–1.5555322.65599090.17162890.7155546No significant changehsa-miR-6729-5p–1.5903272.6585340.17256840.7155546No significant changehsa-miR-3164–1.408142.5424410.17479910.7155546No significant changehsa-miR-4686–2.7121781.76712510.17548590.7155546No significant changehsa-miR-199a-5p–0.8083275.1437260.17663250.7155546No significant changehsa-let-7i-5p0.670520313.7323160.17673540.7155546No significant changehsa-miR-98-5p1.00034125.43526180.17751890.7155546No significant changehsa-miR-148a-3p0.943564616.2903350.17888680.7166447No significant changehsa-miR-424-5p1.89402642.21987140.18088030.7189678No significant changehsa-miR-4738-3p–0.9278434.94219830.18166870.7189678No significant changehsa-miR-548av-5p1.80259982.57862820.18452220.7226147No significant changehsa-miR-548k1.80266042.57862820.18509140.7226147No significant changehsa-miR-10b-5p0.986576313.4901420.18694940.7226147No significant changehsa-miR-6894-5p–2.4249551.62339220.18742970.7226147No significant changehsa-miR-502-3p0.79055246.72115640.18812330.7226147No significant changehsa-miR-548d-5p1.2438272.86000410.19065090.728041No significant changehsa-miR-197-5p2.56812161.55695660.1938980.7361359No significant changehsa-miR-3177-3p–0.8276884.38284170.20480540.7701355No significant changehsa-miR-483-3p–1.9744562.22118160.20521220.7701355No significant changehsa-miR-44431.4411882.01313360.2090350.7769258No significant changehsa-miR-450b-5p1.27731843.62551820.20940110.7769258No significant changehsa-miR-3944-5p–1.9172861.66162470.21246540.7809496No significant changehsa-miR-378a-3p–0.69355313.5292090.21345820.7809496No significant changehsa-miR-1268b–1.2537012.8354880.21407350.7809496No significant changehsa-miR-1301-3p–0.6724486.39650390.21531590.7811183No significant changehsa-miR-23b-3p0.83172056.71901260.21667840.7817181No significant changehsa-let-7f-5p0.981304412.7717150.22033570.7827953No significant changehsa-miR-323a-3p1.54215092.9337770.22068170.7827953No significant changehsa-miR-146a-3p–1.4717242.83360030.22298310.7827953No significant changehsa-miR-3615–0.56001711.590870.22315730.7827953No significant changehsa-miR-3653-3p1.78846592.89437710.226650.7827953No significant changehsa-miR-4750-5p2.19884622.28826480.22701040.7827953No significant changehsa-miR-30b-5p–1.1739842.81233670.22811710.7827953No significant changehsa-miR-4685-3p1.70021632.35987040.22841970.7827953No significant changehsa-miR-29a-3p0.62965727.10611010.22857090.7827953No significant changehsa-miR-338-5p0.98310615.15412940.22896460.7827953No significant changehsa-miR-409-5p–2.4550291.86603020.23039490.7835825No significant changehsa-miR-6735-5p–1.1133512.72528620.23586950.7980454No significant changehsa-miR-451a0.69226917.3080790.23857710.8030457No significant changehsa-miR-543–0.7868275.84795380.24408160.8039067No significant changehsa-miR-320a–0.81534217.4506790.2448790.8039067No significant changehsa-miR-9-5p–1.3712763.5878570.24594020.8039067No significant changehsa-miR-4669–1.5951445.86937760.24634950.8039067No significant changehsa-miR-423-5p0.942799214.8185820.2474310.8039067No significant changehsa-miR-505-5p–1.2712113.17836290.24833590.8039067No significant changehsa-miR-433-3p–0.8475413.96455980.24948750.8039067No significant changehsa-miR-378f–0.7110485.40205750.25067520.8039067No significant changehsa-miR-369-5p1.48647542.46927210.25106320.8039067No significant changehsa-miR-2355-3p–1.150723.77394930.25114390.8039067No significant changehsa-miR-203a-3p–1.0094012.95502790.25243440.8040958No significant changehsa-miR-324-5p–1.1598042.78222150.25395810.8050225No significant changehsa-miR-6866-5p1.215842.67713940.25869370.8156615No significant changehsa-miR-146b-5p–0.5062910.0583820.25981260.8156615No significant changehsa-miR-6850-5p–1.3009323.58139080.26217240.8191319No significant changehsa-miR-6805-5p1.6855671.92144730.26607180.8253837No significant changehsa-miR-1288-3p0.71954333.78221210.26981330.8253837No significant changehsa-miR-671-5p–1.325282.76357740.26984850.8253837No significant changehsa-miR-31981.14730642.83459480.27072230.8253837No significant changehsa-let-7g-5p0.684577311.3755670.27323240.8253837No significant changehsa-miR-15a-5p1.19489873.68488850.27407720.8253837No significant changehsa-miR-125b-5p–0.7320387.84266420.27418470.8253837No significant changehsa-miR-1304-3p0.90435262.91665870.27807660.8253837No significant changehsa-miR-1307-5p0.90638195.22430530.27963020.8253837No significant changehsa-miR-1304-5p0.77626744.53056690.28023730.8253837No significant changehsa-let-7a-3p0.93309174.81543460.28309360.8253837No significant changehsa-let-7c-5p0.68757798.81675550.28614860.8253837No significant changehsa-miR-501-3p0.50044238.05750920.28676480.8253837No significant changehsa-miR-45041.03506082.53260260.28725580.8253837No significant changehsa-miR-6740-5p–1.5811231.73439980.28855720.8253837No significant changehsa-miR-548ah-3p–1.3043423.31975060.28907540.8253837No significant changehsa-miR-548ah-5p1.92948961.70389120.28935560.8253837No significant changehsa-miR-483-5p–0.9305617.5649090.29140310.8253837No significant changehsa-miR-660-5p0.70754084.28822360.29142570.8253837No significant changehsa-miR-21-3p0.95423212.41572150.29314740.8253837No significant changehsa-miR-3934-5p–1.2127192.70025450.29332180.8253837No significant changehsa-miR-636–0.9648913.85631120.29356470.8253837No significant changehsa-miR-548o-3p0.6819336.07712910.2939880.8253837No significant changehsa-miR-6847-5p–1.8498931.8003030.2945090.8253837No significant changehsa-miR-548e-3p1.68746982.81395970.29584370.825581No significant changehsa-miR-548aq-3p1.06581742.53636060.29885530.8304362No significant changehsa-miR-548p–1.2827762.57488820.30251630.8347429No significant changehsa-miR-224-5p–0.8157625.42590380.30296180.8347429No significant changehsa-miR-6842-3p–0.540626.41819740.306620.8353266No significant changehsa-miR-6842-5p–1.101073.13502160.30670380.8353266No significant changehsa-miR-664a-5p–0.5116497.56910250.30701130.8353266No significant changehsa-miR-29b-3p0.91357123.38570840.31041690.8410881No significant changehsa-miR-47921.62819131.76705140.31320490.8451357No significant changehsa-miR-374b-5p1.14942072.79769080.31617450.8482237No significant changehsa-miR-548ar-3p2.03686421.72913850.31736620.8482237No significant changehsa-miR-101-3p0.509177712.222810.31904730.8482237No significant changehsa-miR-19b-3p–0.5136617.86911280.31954520.8482237No significant changehsa-miR-138-5p1.7468352.94301730.32225190.8519453No significant changehsa-miR-127-5p1.33999191.93871150.32382210.8526446No significant changehsa-miR-6767-5p–1.2154242.66487080.32608740.8551609No significant changehsa-miR-378g–0.6752234.94050710.32963510.8602968No significant changehsa-miR-1291–1.0524213.06565290.33143690.8602968No significant changehsa-miR-340-5p0.59215067.14479210.33199820.8602968No significant changehsa-miR-27b-3p0.56060849.23279370.33542140.865732No significant changehsa-miR-133a-3p–1.0105862.7950730.33823560.8695584No significant changehsa-miR-3180-3p–0.8465833.85980080.34111220.8701843No significant changehsa-miR-3180–0.8465823.85980080.34114420.8701843No significant changehsa-miR-548f-3p1.06617863.27496080.34815260.8846056No significant changehsa-miR-454-3p1.57228451.8749250.35276710.8870067No significant changehsa-miR-3605-5p0.82218273.58290530.35285110.8870067No significant changehsa-miR-36560.95755666.4010010.35414520.8870067No significant changehsa-miR-200c-3p–0.7122894.396080.35503010.8870067No significant changehsa-miR-4668-5p1.3920881.62407170.35588940.8870067No significant changehsa-miR-125a-3p0.53996495.17150310.35929550.8870821No significant changehsa-miR-44491.59545641.93204230.35945720.8870821No significant changehsa-miR-409-3p–0.5432169.76194620.36136280.8870821No significant changehsa-miR-191-5p–0.43383810.6199170.36161480.8870821No significant changehsa-miR-378b–0.8808432.20214480.3638230.8870821No significant changehsa-miR-7704–0.9841223.39577060.36414250.8870821No significant changehsa-miR-221-5p–1.0027643.49541760.36739520.8870821No significant changehsa-miR-25-5p–0.4931495.60918140.36794070.8870821No significant changehsa-miR-425-3p0.5418835.31046130.36814590.8870821No significant changehsa-miR-4533–1.7605732.12541060.37040920.8888605No significant changehsa-miR-933–0.9591652.33753050.37530030.8888605No significant changehsa-let-7a-5p0.649732212.9758140.37534050.8888605No significant changehsa-miR-4470–0.8925242.81220580.376950.8888605No significant changehsa-miR-2278–1.7666991.87787930.37704780.8888605No significant changehsa-miR-486-5p–0.45793916.6860460.38320730.8888605No significant changehsa-miR-3940-3p–1.4080481.66087320.38328430.8888605No significant changehsa-miR-3187-3p–0.8921372.59216950.38557470.8888605No significant changehsa-miR-44970.73927053.60146120.38708670.8888605No significant changehsa-miR-199b-5p0.98364113.06200570.38806510.8888605No significant changehsa-miR-619-5p0.79148853.69301730.39191010.8888605No significant changehsa-miR-486-3p–0.5271639.29471090.39220850.8888605No significant changehsa-miR-877-5p0.8871422.78355250.39366750.8888605No significant changehsa-miR-26a-5p–0.43852612.0349810.39416450.8888605No significant changehsa-miR-130a-3p0.79318393.4524870.39622620.8888605No significant changehsa-miR-5189-3p–1.8777461.58503350.39793360.8888605No significant changehsa-miR-219a-1-3p0.61665563.92891590.39856450.8888605No significant changehsa-miR-3613-5p1.04868123.43537810.39904070.8888605No significant changehsa-miR-6868-3p1.57841151.832750.39931110.8888605No significant changehsa-miR-43260.50593124.6958750.39969360.8888605No significant changehsa-miR-142-3p–0.5399925.27611150.39995010.8888605No significant changehsa-miR-188-5p–1.7720541.83302910.40037390.8888605No significant changehsa-miR-628-3p–0.8154033.53649410.40201940.8888605No significant changehsa-miR-3163–1.7724861.73363990.40391950.8888605No significant changehsa-miR-6734-5p–0.7852772.78965610.40393290.8888605No significant changehsa-miR-450a-2-3p1.43172921.75965010.4049950.8888605No significant changehsa-miR-181a-2-3p0.50759865.43326870.40736370.8888605No significant changehsa-miR-95-3p0.85445313.15195620.40839020.8888605No significant changehsa-miR-197-3p–0.4494935.82764890.40953430.8888605No significant changehsa-let-7d-3p–0.567878.82932060.4098230.8888605No significant changehsa-miR-1343-3p–0.9465812.51332190.4110810.8888605No significant changehsa-miR-28-5p–0.6213884.6300080.41748680.8997323No significant changehsa-miR-4746-5p–0.4868394.76261530.42122210.9022778No significant changehsa-miR-766-5p–0.7376943.37272110.4220330.9022778No significant changehsa-miR-215-5p–0.4806047.11617660.42495660.9022778No significant changehsa-miR-5584-5p–1.3797731.78465950.4250090.9022778No significant changehsa-let-7e-5p–0.463965.72918920.42678140.9022778No significant changehsa-miR-204-5p1.05626793.14460460.42808740.9022778No significant changehsa-miR-505-3p1.01153242.20079870.43111190.9022778No significant changehsa-miR-3960–0.8203233.02524960.43125140.9022778No significant changehsa-miR-769-5p0.41313476.3285320.4341830.9022778No significant changehsa-miR-574-5p0.51653054.41206080.43461840.9022778No significant changehsa-miR-4745-5p1.10351721.88345070.43517740.9022778No significant changehsa-miR-6826-5p–1.5498231.66799060.43742850.9022778No significant changehsa-miR-6873-3p–1.8542911.84344490.43753160.9022778No significant changehsa-miR-1255a–1.4927191.84635840.43891870.9022778No significant changehsa-miR-196a-5p1.58973581.72112080.43939410.9022778No significant changehsa-miR-541-3p1.65964731.5370370.44331410.9048498No significant changehsa-miR-411-5p–1.0765072.20874150.44351090.9048498No significant changehsa-miR-5010-3p–0.8832362.71594180.44480370.9048498No significant changehsa-miR-627-5p–0.9185253.62527890.44661780.9057188No significant changehsa-miR-1285-3p0.39455647.18308840.45172630.9132424No significant changehsa-miR-190b–0.9639082.6771740.45450070.9144084No significant changehsa-miR-100-5p0.42004139.59219160.45600860.9144084No significant changehsa-miR-361-5p0.5350134.80838030.45650410.9144084No significant changehsa-miR-5187-5p–0.8674992.90178730.45915330.9148355No significant changehsa-miR-147b–0.5893514.46677790.46233710.9148355No significant changehsa-miR-937-3p–0.9743851.93539140.4643010.9148355No significant changehsa-miR-6510-3p1.01742611.76192310.46441560.9148355No significant changehsa-miR-939-5p–0.6972564.09958990.46512440.9148355No significant changehsa-miR-378h–0.7916172.32606910.46587310.9148355No significant changehsa-miR-2171.30214981.90501080.46652410.9148355No significant changehsa-miR-7641–0.4680294.93669520.46901130.9154936No significant changehsa-miR-548av-3p0.471146.2522220.47335180.9154936No significant changehsa-miR-6741-5p0.59492074.17143290.4753080.9154936No significant changehsa-miR-654-5p–0.4645144.84001680.47639950.9154936No significant changehsa-miR-200b-3p0.70911445.01076980.47660570.9154936No significant changehsa-miR-1255b-5p–0.4513045.60569010.47792320.9154936No significant changehsa-miR-491-5p0.93131272.24976780.47894450.9154936No significant changehsa-miR-25-3p0.348973614.278370.47910030.9154936No significant changehsa-miR-941–0.4094368.26144030.4802180.9154936No significant changehsa-miR-12780.74814352.83171470.48087950.9154936No significant changehsa-miR-136-3p1.13335582.0802220.48562820.9218466No significant changehsa-miR-651-5p0.82967412.88578990.48939420.9261983No significant changehsa-miR-2115-5p–0.6541343.85682410.49471840.9261983No significant changehsa-miR-125b-1-3p–0.7100742.58218460.49518710.9261983No significant changehsa-miR-432-5p0.5194595.77323120.49737340.9261983No significant changehsa-miR-370-3p–0.4492918.52314890.49955610.9261983No significant changehsa-miR-30e-3p0.43030926.20127460.50154770.9261983No significant changehsa-miR-4664-3p0.69642192.92129320.50269160.9261983No significant changehsa-miR-199b-3p0.39839557.74128340.50287770.9261983No significant changehsa-miR-199a-3p0.39839577.74128340.50291590.9261983No significant changehsa-miR-3135a0.847272.2651360.5032140.9261983No significant changehsa-miR-200b-5p–1.0187362.43453790.50418330.9261983No significant changehsa-miR-485-5p–0.4525215.31785140.50578170.9261983No significant changehsa-miR-7854-3p0.56683724.12618320.50635950.9261983No significant changehsa-miR-6500-3p1.84579381.57899950.51020980.9267524No significant changehsa-miR-342-5p–0.3711017.97578910.51177930.9267524No significant changehsa-miR-4429–0.4869076.55450030.51282560.9267524No significant changehsa-miR-106b-3p0.29716589.69627240.51355980.9267524No significant changehsa-miR-1468-5p–0.431494.43149040.51388610.9267524No significant changehsa-miR-192-5p–0.38926810.9070.51517790.9267524No significant changehsa-miR-30b-3p–0.7486552.23924320.51997140.9326946No significant changehsa-miR-200a-3p0.60847556.00343140.52255110.9326946No significant changehsa-miR-99b-3p0.44259545.29623110.52420530.9326946No significant changehsa-miR-125b-2-3p–0.4959674.20761810.52525650.9326946No significant changehsa-miR-128-3p–0.41372211.3839020.52623590.9326946No significant changehsa-miR-92b-3p–0.4328075.47865380.5270510.9326946No significant changehsa-miR-143-5p0.62690913.08705240.52976080.9335995No significant changehsa-miR-39110.93204732.74157640.53042180.9335995No significant changehsa-miR-103a-3p0.305439110.3547310.5324890.933819No significant changehsa-miR-30c-2-3p0.84254852.17526550.53544840.933819No significant changehsa-miR-495-3p0.52917275.46755320.53645520.933819No significant changehsa-miR-26a-2-3p1.51988741.60293440.5375990.933819No significant changehsa-miR-126-3p–0.30727312.4131480.53947430.933819No significant changehsa-miR-3682-3p0.81435842.10604390.54073250.933819No significant changehsa-miR-4448–0.8761611.56481660.54325240.933819No significant changehsa-miR-132-5p0.42505664.38400810.54362330.933819No significant changehsa-miR-93-3p–0.5913152.84805140.54454390.933819No significant changehsa-miR-32-3p1.43284081.7613570.54970120.933819No significant changehsa-miR-20a-5p0.37788387.20584720.55027760.933819No significant changehsa-let-7i-3p–1.3222391.87907770.55057620.933819No significant changehsa-miR-329-3p1.00609241.84287530.55172860.933819No significant changehsa-miR-493-5p1.09141512.07293940.55279740.933819No significant changehsa-miR-6878-5p–0.962071.82759460.55472830.933819No significant changehsa-miR-151a-3p–0.2703915.0426130.55475180.933819No significant changehsa-miR-92a-1-5p–0.9316992.28044790.55630170.933819No significant changehsa-miR-46600.95677592.22838560.55760350.933819No significant changehsa-miR-4753-5p0.81886792.33542970.55771730.933819No significant changehsa-miR-340-3p–0.5254084.26399040.56195480.9344595No significant changehsa-miR-34c-5p–0.9097241.66551840.56218010.9344595No significant changehsa-miR-10b-3p0.46651914.88836030.56239290.9344595No significant changehsa-miR-4661-5p0.52124493.07500910.57075370.9441812No significant changehsa-miR-32-5p0.56113413.12758750.57113570.9441812No significant changehsa-miR-452-5p–0.3587466.02193010.57335420.9444801No significant changehsa-miR-548j-3p–0.7538822.61906970.57421830.9444801No significant changehsa-miR-148b-5p–0.6656222.46644560.57565550.9444801No significant changehsa-miR-1180-3p–0.274829.02817030.57748780.9448464No significant changehsa-miR-17-3p–0.9751261.89032050.57877270.9448464No significant changehsa-miR-3127-5p0.58021732.63557280.58319930.9496986No significant changehsa-miR-195-5p0.74328782.50186880.58822750.9555039No significant changehsa-miR-6515-5p0.4809453.63037550.59777440.9686022No significant changehsa-miR-12940.34636514.93233710.60059490.9707635No significant changehsa-miR-6514-5p0.61986552.40062830.60286920.9720336No significant changehsa-miR-7850-5p–1.0244762.14573570.6046710.9725373No significant changehsa-miR-132-3p–0.5232323.06405870.60917890.9773803No significant changehsa-miR-5581-3p–1.1186941.70233920.61087620.9777013No significant changehsa-miR-6786-3p–0.8200011.85401610.61536520.9824779No significant changehsa-miR-24-1-5p–0.5887332.49939690.6172750.9831234No significant changehsa-miR-296-3p0.61981842.70346050.62007190.9851751No significant changehsa-miR-598-3p0.67287582.25668530.62366780.9884833No significant changehsa-miR-4646-5p–0.5408472.29127290.62627760.9902161No significant changehsa-miR-4440–0.5901122.27935660.63890040.998528No significant changehsa-miR-1273 h-5p–0.4239213.7907860.63914110.998528No significant changehsa-miR-576-3p–0.2376878.73406720.64040480.998528No significant changehsa-miR-45160.53847991.82707860.64440290.998528No significant changehsa-miR-28-3p–0.2163849.23620650.64630920.998528No significant changehsa-miR-140-5p0.36915164.34508040.64755970.998528No significant changehsa-miR-4781-3p–0.7666541.96005320.64788830.998528No significant changehsa-miR-320b–0.31876313.643760.64908340.998528No significant changehsa-miR-4726-5p–1.1150351.93218520.65063760.998528No significant changehsa-miR-615-3p0.49955485.29575990.65134570.998528No significant changehsa-miR-44371.12385671.70133770.65213910.998528No significant changehsa-miR-185-3p–0.2755835.40417650.65470880.998528No significant changehsa-miR-1250-5p–0.4602753.12922870.65726770.998528No significant changehsa-miR-532-5p0.202434210.4635050.65728830.998528No significant changehsa-miR-3620-5p0.53274812.17305180.65812880.998528No significant changehsa-miR-6862-5p0.73397341.65785420.65835560.998528No significant changehsa-miR-339-5p0.35993483.54203690.66013040.998528No significant changehsa-miR-193b-3p0.83553951.83482190.66035390.998528No significant changehsa-miR-24-3p0.211912712.4836150.66194220.998528No significant changehsa-miR-361-3p0.19673878.92902520.66303330.998528No significant changehsa-miR-4487–1.1508091.56979530.66367380.998528No significant changehsa-miR-548 am-3p0.51512262.96106650.66725880.998528No significant changehsa-miR-335-3p0.40410493.41631320.66996150.998528No significant changehsa-miR-4840.2296378.23030320.67010360.998528No significant changehsa-miR-320d0.32037038.77208330.67086870.998528No significant changehsa-let-7d-5p0.27110849.26645150.67169540.998528No significant changehsa-miR-196b-5p0.43297912.75833640.67675310.998528No significant changehsa-miR-27b-5p0.5585642.79125710.67888340.998528No significant changehsa-miR-1287-5p0.36756213.72488610.68083770.998528No significant changehsa-miR-92a-3p–0.25264713.8377650.68251840.998528No significant changehsa-miR-1273c–0.5525222.47816650.68296090.998528No significant changehsa-miR-22-3p–0.18138912.4470270.68533610.998528No significant changehsa-miR-766-3p0.56141172.04063060.68555550.998528No significant changehsa-miR-6503-3p–0.3002073.79723930.68607940.998528No significant changehsa-miR-1285-5p0.99502261.64543970.68648920.998528No significant changehsa-miR-181c-3p0.42928272.69015740.68658360.998528No significant changehsa-miR-44670.5934212.53656320.69027731No significant changehsa-miR-642a-3p–0.4754323.2181890.69403721No significant changehsa-miR-3605-3p–0.6591322.47707230.69592271No significant changehsa-miR-3120-3p–0.3252193.14885790.69868961No significant changehsa-miR-1273d–0.3907832.85242220.70065831No significant changehsa-miR-222-3p0.20871158.7331340.70301741No significant changehsa-miR-181b-5p0.22306567.82561680.70937481No significant changehsa-miR-45080.285009710.4247080.71041691No significant changehsa-miR-548u–0.5643021.98144850.71057771No significant changehsa-miR-99b-5p0.19506439.44669710.71232011No significant changehsa-miR-12610.62356881.68847820.71264221No significant changehsa-miR-381-3p–0.2306755.28898090.71600371No significant changehsa-miR-504-5p0.65952511.8402550.71635791No significant changehsa-let-7f-1-3p–0.4560752.67975660.72479341No significant changehsa-miR-18a-3p–0.4437583.33363270.72603521No significant changehsa-miR-6180.41278252.98917590.7269831No significant changehsa-miR-3138–0.749131.82681810.73098191No significant changehsa-miR-328-3p–0.1721316.75136950.73203521No significant changehsa-miR-6806-3p0.37067073.26658150.73294871No significant changehsa-miR-144-3p–0.1901365.92845790.73395611No significant changehsa-miR-12700.5207362.00350590.73600311No significant changehsa-miR-4659b-3p–0.9173161.59456080.7363331No significant changehsa-miR-514a-3p0.66445751.76271730.73646871No significant changehsa-miR-4654–0.6395382.28145970.74282391No significant changehsa-miR-79760.22802994.05529380.74313021No significant changehsa-miR-873-3p0.29947084.22328170.74610151No significant changehsa-miR-424-3p0.17113237.2134380.74663181No significant changehsa-miR-942-3p–0.570662.27594550.75202991No significant changehsa-miR-214-5p0.4314212.53257430.75351771No significant changehsa-miR-610–0.3835322.74638280.753931No significant changehsa-miR-186-5p–0.14693910.2104790.75535951No significant changehsa-miR-629-5p0.169518911.2647910.75701341No significant changehsa-miR-205-5p–0.4554822.21632670.75750521No significant changehsa-miR-1538–0.3778362.47731780.75819751No significant changehsa-miR-4742-3p–0.5744251.96564150.76165091No significant changehsa-miR-152-3p–0.1747237.87559730.76390431No significant changehsa-miR-1260b–0.2319884.02587880.7648171No significant changehsa-miR-191-3p–0.258573.611250.76625291No significant changehsa-miR-12900.17213728.29027430.76720011No significant changehsa-miR-3158-3p–0.15083410.6370150.76726921No significant changehsa-miR-128-1-5p0.50004032.07667890.76826681No significant changehsa-miR-42860.51626471.84431750.77484521No significant changehsa-miR-335-5p0.19355975.83161060.77602721No significant changehsa-miR-210-3p–0.1962575.32674140.77678071No significant changehsa-miR-26b-5p0.17819767.66481120.7790131No significant changehsa-miR-7-5p0.133792311.0934450.78296571No significant changehsa-miR-3691-5p0.3796012.19074580.78578471No significant changehsa-miR-99a-5p–0.15037213.5698670.78886861No significant changehsa-miR-183-5p0.17127028.48130480.79175231No significant changehsa-miR-6821-5p–0.4926811.82365720.79971481No significant changehsa-miR-6750-5p–0.2565612.97910130.80274271No significant changehsa-miR-18a-5p0.53120051.90859350.80392161No significant changehsa-miR-3679-5p–0.500481.96578910.80706121No significant changehsa-miR-345-5p–0.1260437.11035770.80757581No significant changehsa-miR-155-5p0.18976885.32611390.80804941No significant changehsa-miR-659-5p–0.1767874.48250360.80898721No significant changehsa-miR-4772-5p–0.3840252.21723990.81238321No significant changehsa-miR-576-5p–0.3178233.44461280.81606071No significant changehsa-miR-6829-5p0.65732331.68035280.81622841No significant changehsa-miR-5189-5p–0.2647123.69693120.81978241No significant changehsa-miR-4797-3p0.49772171.57064560.8207531No significant changehsa-miR-378d0.15987436.47448160.82218081No significant changehsa-miR-151a-5p–0.189853.77195240.82317261No significant changehsa-miR-320c–0.15948610.9179390.82353171No significant changehsa-miR-106b-5p–0.1385435.33410270.8242241No significant changehsa-miR-379-5p0.2220084.58389070.82613511No significant changehsa-miR-20b-5p–0.138324.86495190.82702141No significant changehsa-miR-1273f0.27242472.5875970.82929211No significant changehsa-let-7b-5p–0.1078413.3251420.82992651No significant changehsa-miR-3140-3p–0.3521472.09327230.83086511No significant changehsa-miR-145-5p0.17944924.35112820.83211141No significant changehsa-miR-548ap-5p0.26004832.0135020.83254451No significant changehsa-miR-181a-3p–0.2107373.05544960.83339321No significant changehsa-miR-378i–0.121088.79994960.83569991No significant changehsa-miR-3199–0.4643731.99238270.83750081No significant changehsa-miR-3127-3p–0.6228971.80054860.8398741No significant changehsa-miR-429–0.3491532.55108790.84538631No significant changehsa-miR-374a-5p0.22645043.089390.84936381No significant changehsa-miR-382-5p0.10952126.94169820.85134191No significant changehsa-miR-6761-5p0.6236791.54953840.8515641No significant changehsa-miR-6875-5p0.52116681.94003190.85157981No significant changehsa-miR-423-3p–0.08161111.8724070.85511711No significant changehsa-miR-60870.58226171.60758620.85540711No significant changehsa-miR-6882-5p0.51914951.57871260.85633171No significant changehsa-miR-3196–0.5497641.56109050.85973651No significant changehsa-miR-96-5p0.20175213.50279270.86046981No significant changehsa-miR-339-3p–0.1011567.38903870.86105931No significant changehsa-miR-30e-5p–0.0801269.47713370.86192781No significant changehsa-miR-23a-5p–0.1522713.54463310.86531281No significant changehsa-miR-46570.29775582.20538270.86551211No significant changehsa-miR-4741–0.3336541.77722740.86783811No significant changehsa-miR-574-3p–0.1379524.59919810.86852751No significant changehsa-miR-493-3p–0.12035.61544890.8692641No significant changehsa-miR-455-5p–0.1574624.05151210.87127331No significant changehsa-miR-4747-5p–0.6615221.75324460.87360841No significant changehsa-miR-31790.67397581.53721340.87393231No significant changehsa-miR-374a-3p0.30153781.71773520.87806221No significant changehsa-miR-378a-5p0.29666642.44214760.87911341No significant changehsa-miR-760–0.0938725.95053120.88034341No significant changehsa-miR-4639-5p–0.4731421.60209460.88136411No significant changehsa-miR-31760.14166032.6330190.88299531No significant changehsa-miR-3124-5p–0.1733893.12592510.88445191No significant changehsa-miR-625-3p–0.1070026.39897750.88452031No significant changehsa-miR-3150b-3p–0.095925.01805380.88502721No significant changehsa-miR-4466–0.2519131.82755560.88572441No significant changehsa-miR-106a-5p0.14048813.92838780.88758241No significant changehsa-miR-24-2-5p–0.0760835.91032130.88842911No significant changehsa-miR-12540.14209863.16687220.8891561No significant changehsa-miR-10a-5p–0.09212711.2620340.88920131No significant changehsa-miR-17-5p–0.0787096.52551910.88968581No significant changehsa-miR-330-5p–0.0828625.91351670.8897761No significant changehsa-miR-3074-5p0.29563171.90121050.89003591No significant changehsa-miR-136-5p0.18043243.12087970.89618271No significant changehsa-miR-324-3p–0.1335593.00588110.89999551No significant changehsa-miR-550a-3-5p0.06422576.57963740.90140261No significant changehsa-miR-26b-3p–0.3079552.00101250.90211221No significant changehsa-miR-4498–0.3345581.74913140.90611821No significant changehsa-miR-331-3p0.27047662.01276790.90779981No significant changehsa-miR-579-5p–0.2454582.63888520.90808541No significant changehsa-miR-195-3p0.10677414.20953030.91043021No significant changehsa-miR-365b-5p0.27076651.95952750.91052691No significant changehsa-miR-3909–0.3078871.97039240.91471021No significant changehsa-miR-146a-5p–0.04981312.1375340.9152021No significant changehsa-miR-3064-5p0.23253212.11061260.91633361No significant changehsa-miR-625-5p0.36274391.89578840.91653561No significant changehsa-miR-499a-5p0.06958464.79891120.91835911No significant changehsa-miR-548q–0.430381.81649820.91986131No significant changehsa-miR-5009-5p–0.0852353.92493930.92116421No significant changehsa-miR-44730.18097761.96477360.92291541No significant changehsa-miR-616-3p–0.1486832.35644930.92511341No significant changehsa-miR-1260a0.12456253.38670940.92682891No significant changehsa-miR-127-3p–0.0573087.70988320.92801091No significant changehsa-miR-10a-3p0.2019662.41650850.92851911No significant changehsa-miR-942-5p–0.0505385.75541590.93040111No significant changehsa-miR-3150a-5p–0.2480332.18404620.93334781No significant changehsa-miR-5010-5p–0.1896892.3508880.93451511No significant changehsa-miR-19a-3p0.06687185.54691390.93496231No significant changehsa-miR-46470.10210533.75807730.93614561No significant changehsa-miR-30c-5p–0.05086.84280240.9376091No significant changehsa-miR-221-3p0.03994298.24074370.93922161No significant changehsa-miR-30d-5p–0.0373412.7887920.94175391No significant changehsa-miR-200a-5p0.21007912.9142630.94178151No significant changehsa-miR-4732-5p–0.0493046.26717870.94291881No significant changehsa-miR-372-3p0.12243693.88806730.94389891No significant changehsa-miR-93-5p–0.037247.5797070.94480461No significant changehsa-miR-550a-5p0.03950626.15471690.94749481No significant changehsa-miR-21-5p–0.03049511.735240.9477891No significant changehsa-miR-342-3p0.04795214.65904610.95501051No significant changehsa-miR-23b-5p–0.0682613.88070560.956381No significant changehsa-miR-3190-3p0.13267342.94808060.95714011No significant changehsa-miR-363-5p0.13313892.99216040.95848561No significant changehsa-miR-874-3p–0.0748643.62639680.95932451No significant changehsa-miR-4755-3p–0.0804793.13702780.96040281No significant changehsa-miR-1-3p–0.0364187.17981750.96321191No significant changehsa-miR-126-5p–0.0653054.00844070.96478031No significant changehsa-miR-652-3p–0.0293455.91222460.96635231No significant changehsa-miR-129-5p–0.0282946.98213230.96753281No significant changehsa-miR-16-2-3p–0.0275238.8119070.96832081No significant changehsa-miR-1306-3p–0.0314425.77171780.9703661No significant changehsa-miR-92b-5p–0.046484.40999620.97245631No significant changehsa-miR-194-5p0.0221137.91644170.97273061No significant changehsa-let-7b-3p0.04378953.48389390.97349181No significant changehsa-miR-130b-3p–0.0258124.15043860.97718691No significant changehsa-miR-584-5p–0.0150479.19855010.97744081No significant changehsa-miR-151b0.02436524.21123580.97873141No significant changehsa-miR-181a-5p–0.01028210.9394460.98200321No significant changehsa-miR-6852-5p0.0188144.59476720.98626411No significant changehsa-miR-425-5p0.00625138.96398750.98960491No significant changehsa-miR-181d-5p–0.0086515.018253611No significant changehsa-miR-516b-5p–0.1026282.819071111No significant changehsa-miR-641–0.0978432.270386811No significant changehsa-miR-1273a–0.3421871.540993611No significant changehsa-miR-548j-5p0.22381241.935619411No significant changehsa-miR-548az-5p0.1321752.232959811No significant changehsa-miR-371b-5p0.34747161.600509711No significant changehsa-miR-6716-3p0.33155851.592339811No significant changehsa-miR-4306–0.3288551.590447911No significant changehsa-miR-6839-5p0.29734051.692811111No significant changehsa-miR-6774-5p–0.2536021.540093811No significant changehsa-miR-6818-5p–0.2347821.88261811No significant changehsa-miR-6781-5p0.22342651.536186811No significant changehsa-miR-4800-5p0.21543582.014541411No significant changehsa-miR-365a-5p0.18276951.777959911No significant changehsa-miR-500b-3p0.16435852.039601811No significant changehsa-miR-5810.16378641.996926211No significant changehsa-miR-6501-5p0.14223511.706718411No significant changehsa-miR-494-3p–0.1318721.77249511No significant changehsa-miR-556-5p–0.130521.738435811No significant changehsa-miR-6780a-5p0.12351971.98260411No significant changehsa-miR-6815-5p–0.0749962.433705111No significant changehsa-miR-50960.07287142.329442311No significant changehsa-miR-5003-3p–0.0705532.037079511No significant changehsa-miR-4724-5p0.06905481.704881911No significant changehsa-miR-431-5p0.06336662.275235511No significant changehsa-miR-664b-5p–0.0622522.053089311No significant changehsa-miR-3122–0.0607051.737748311No significant changehsa-miR-36900.04404393.098556811No significant changehsa-miR-30c-1-3p–0.0379411.868458811No significant changehsa-miR-3675-5p0.03773251.912371111No significant changehsa-miR-6733-5p–0.0242921.594861811No significant changehsa-miR-4433b-3p–0.0148774.828172211No significant changehsa-miR-412-5p–0.0052982.410213511No significant changehsa-miR-29c-5p–0.0011972.704549311No significant changehsa-miR-144-5p–0.0002544.026345111No significant changehsa-miR-375–2.90792810.6212114.83E-083.16E-05Downhsa-miR-885-3p–5.2739852.40018320.00123260.2345272Downhsa-miR-122-5p–2.15982117.226280.00255470.2345272Downhsa-miR-4433b-5p–2.2743284.52276630.00266410.2345272Downhsa-miR-4446-3p–1.7113384.69007480.00704620.2367778Downhsa-miR-885-5p–4.1349222.50153120.00840430.2494538Downhsa-miR-362-5p–4.250442.11849630.01005740.2623126Downhsa-miR-150-3p–1.4622536.40668430.0104930.2623126Downhsa-miR-3173-5p–2.2546042.67844230.01692410.3683817Downhsa-miR-139-3p–1.4297496.62995380.01826710.3847879Downhsa-miR-378e–2.9475713.60081840.02176260.4111132Downhsa-miR-193b-5p–1.5259967.07637640.02203520.4111132Downhsa-miR-4435–2.0385392.53926220.02392510.4123441Downhsa-miR-3191-3p–3.1123642.26003130.03244870.4546377Downhsa-miR-4665-5p–1.7863674.01037890.03519450.4787914Downhsa-miR-1249-3p–4.3666911.63614650.03677610.4900979Downhsa-miR-1299–2.2319418.51187650.03775540.4914672Downhsa-miR-6884-5p–1.815154.26705090.03913670.4914672Downhsa-miR-1247-5p–4.906651.87845320.04075620.5021471Downhsa-miR-4732-3p–1.1080726.60872680.042290.5113961Downhsa-miR-320e–1.9942685.44740590.04551230.5261696Downhsa-miR-146b-3p–1.4064134.61727670.0466920.5261696Downhsa-miR-6511a-5p–3.2617562.13367660.04673480.5261696Downhsa-miR-193a-5p–1.20292611.1551220.04893390.5415905DownFigure 1
Scanning differentially expressed miRNAs. (a) Differentially expressed miRNAs in GSE199942. |LogFC| >1 andP value <0.05 were set to screen. (b) Heatmap of the differentially expressed miRNAs in GSE199942. (c) Venn diagram representing the intersection of up-regulated expressed miRNAs in the 2 data sets (GSE199942 and PMC8702168). (d) Venn diagram representing the intersection of down-regulated expressed miRNAs in the 2 data sets (GSE199942 and PMC8702168).
(a)(b)(c)(d)


### 3.2. Construction of miRNA-mRNA Network

Next, we analyzed the potential targets of the three differentially expressed serum microvesicles miRNAs by miRWalk, and found a total of 593 genes may be co-regulated by at least two of the three miRNAs, of which 192 genes were co-targeted by hsa-miR-15b-5p and hsa-miR-16-5p, 250 genes were co-targeted by hsa-miR-15b-5p and hsa-miR-184, 92 genes were co-targeted by hsa-miR-16-5p and hsa-miR-184, and 59 genes were co-targeted by the three miRNAs (Figures2(a) and 2(b) and Table 2). Given that miRNA negatively regulates the expressions of its targets, we further intersected the 593 prediction genes with the downregulated-expressed genes of PBMCs of IS patients from GSE22255 [26], and obtained 25 genes (Figure 2(c)). The expression of the 25 intersected genes was shown in Figure 2(d). Then, the network containing 3 miRNAs and 25 mRNAs was visualized by the “ggallouvial” package of R software and was shown in Figure 3.Figure 2
Construction of miRNA-mRNA Network. (a) The miRNA-target gene network for differentially expressed miRNAs. (b) Venn diagram showing the overlap among the predicted targets of the differentially expressed miRNAs. (c) Venn diagram showing the overlap between down-regulated expressed genes and predicted targets of the differentially expressed miRNAs. (d) Heat map of the 25 intersected genes.
(a)(b)(c)(d)Table 2
Target genes of the three miRNAs.
Potential targets of hsa-miR-15b-5pPotential targets of hsa-miR-16-5pPotential targets of hsa-miR-184Potential targets of hsa-miR-15b-5p AND hsa-miR-16-5p NOT hsa-miR-184Potential targets of hsa-miR-15b-5p AND hsa-miR-184 NOT hsa-miR-16-5pPotential targets of hsa-miR-16-5p AND hsa-miR-184 NOT hsa-miR-15b-5pPotential targets of hsa-miR-15b-5p AND hsa-miR-16-5p AND 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 3
Sankey diagram of the miRNA-mRNA Network in IS.


### 3.3. Enrichment Analysis of Differentially Expressed Genes

To explore the potential biological functions of the above downregulated-expressed 25 mRNAs, we performed GO annotation and KEGG pathway enrichment analysis. The enriched GO annotation included oxidoreduction coenzyme metabolic process, pyridine-containing compound metabolic process, and nicotinamide nucleotide metabolic process in the biological process (BP) category. Sin3 complex, oligosaccharyltransferase complex, and tetraspanin-enriched microdomain were included in the cellular component (CC) category. NAD+ kinase activity, oxidoreductase activity, and beta-1,3-galactosyltransferase activity were included in the molecular function (MF) category (Figure4(a)). KEGG pathway analysis showed these genes were mainly involved in various types of N-glycan biosynthesis, nicotinate and nicotinamide metabolism, and pentose phosphate pathway (Figure 4(b)).Figure 4
GO (a) and KEGG (b) analysis of the 25 intersected genes.
(a)(b)


### 3.4. PPI Network Construction and Validation of Predictive Feature Biomarkers

We further analyzed the protein-protein interaction (PPI) network of the above 25 intersected genes by Genemania and identified the RB binding protein 4 (RBBP4) had the most interactions in the network (Figure5(a)). Furthermore, we performed the LASSO algorithm to narrow down the above 25 intersected genes and identified eight key genes related to IS (Figure 5(b)). The eight genes were BCL11A, RNLS, UBFD1, STT3A, NADK2, GPR26, PPARA, and SAMHD1. Moreover, we used ROC curve to estimate the predictive value of the eight genes, and found that all of the eight genes had a certain diagnostic value in discriminating IS patients from the healthy controls, with an AUC of 0.7 (Figure 5(C)).Figure 5
PPI network construction and validation of predictive feature biomarkers. (a) PPI network of the 25 intersected genes constructed by GENEMANIA. (b) A plot of biomarkers selection by LASSO regression algorithm. (c) ROC diagnostic curve for the eight genes selected by ASSO regression algorithm.
(a)(b)(c)


### 3.5. Immune Cell Patterns in the Peripheral Bloods of IS

Given that peripheral immune system has been shown to play crucial roles in the evolution of ischemic brain damage [27, 28], we further compared the different immune cell patterns of the peripheral bloods between IS patients and normal controls from GSE22255. As shown in Figures 6(a) and 6(b), M0 macrophages and resting mast cells were significantly decreased in the peripheral bloods of IS patients. M1 macrophages and activated mast cells were slightly increased in the IS group but had no significant difference compared to the normal control group (Figure 6(b)). Furthermore, we analyzed the correlation among the immune cells and found that monocytes were significantly positively correlated to M2 macrophages and significantly negatively correlated to activated NK cells and activated mast cells (Figure 7(a)). Moreover, we analyzed the correlation between the immune cells and the eight variables related to IS and found that STT3A was significantly negative-correlated to follicular helper T cells, activated NK cells and resting dendritic cells (Figure 7(b)).Figure 6
Immune cell patterns in the peripheral bloods of IS samples. (a) The composition of immune cells in the peripheral bloods of IS patients. (b) Comparison of the indicated immune cell subtypes between stroke and normal control group.∗P<0.05.
(a)(b)Figure 7
Correlation analysis of immune cells in the peripheral bloods of IS. (a) The correlation between immune cells in IS samples. (b) The correlation between the eight key genes and immune cells. *P<0.05, **P<0.01, ***P<0.001.
(a)(b)


### 3.6. Verification of the Diagnostic Efficacy of the Key miRNAs

GSE117064 were included in this study as the verification series to validate the diagnostic efficacy of the above three miRNAs. Has-miR-16-5p was more advantageous for diagnosing stroke than the other two miRNAs, as shown in Figure8.Figure 8
Verification of the diagnostic efficacy of the key miRNAs. ROC diagnostic curve for the three miRNAs.


## 3.1. Identification of Differentially Expressed Exosomal miRNAs Related to IS

In this study, we analyzed the differentially expressed miRNAs in serumexosomes between IS patients and healthy controls from GSE199942 data sets, and found that 35 upregulated and 24 downregulated miRNAs (Table 1). The expression of these miRNAs is shown in Figures 1(a) and 1(b). Furthermore, the differentially expressed serum exosomes miRNAs were intersected with those differentially expressed miRNAs from freshly removed human stroke brain tissue [25], and three upregulated miRNAs were obtained: hsa-miR-15b-5p, hsa-miR-184, and hsa-miR-16-5p (Figures 1(c) and 1(d)).Table 1
The differentially expressed miRNAs in serum exosomes between IS patients and healthy controls from GSE199942 data sets.
GeneslogFClogCPMP ValueFDRChangehsa-miR-45114.01777513.15193110.0003910.1276672Uphsa-miR-450a-5p2.36085314.750.00187180.2345272Uphsa-miR-15b-5p1.87015655.68244970.00275740.2345272Uphsa-miR-187-3p6.04097993.63702480.00287320.2345272Uphsa-miR-44883.1070225.14011730.00490580.2367778Uphsa-miR-30a-5p2.460329210.2023890.00518250.2367778Uphsa-miR-3688-3p4.51277811.92195090.00548930.2367778Uphsa-miR-30a-3p2.34924787.31374970.00571830.2367778Uphsa-miR-519b-5p5.55156392.28563430.00677650.2367778Uphsa-miR-523-5p5.55156352.28563430.00677690.2367778Uphsa-miR-522-5p5.55152362.28563430.00681490.2367778Uphsa-miR-518e-5p5.551472.28563430.00686640.2367778Uphsa-miR-519c-5p5.55145372.28563430.00688220.2367778Uphsa-miR-519a-5p5.55138482.28563430.00694910.2367778Uphsa-miR-23a-3p1.39001298.91372210.00731520.2367778Uphsa-miR-223-3p1.69909386.59243470.00761460.2367778Uphsa-miR-12624.76793631.84184620.00903330.2564662Uphsa-miR-1843.92838663.48733810.00964320.2623126Uphsa-let-7g-3p4.7771191.83496660.0108460.2623126Uphsa-miR-503-5p1.46372555.97198430.0116940.2727198Uphsa-miR-2115-3p3.35989632.57556350.01412690.3180986Uphsa-miR-365a-3p3.37513443.19586650.02101960.4111132Uphsa-miR-365b-3p3.37521763.19586650.02133120.4111132Uphsa-miR-143-3p1.310620212.3402440.02321670.4123441Uphsa-miR-140-3p1.100650413.623170.02399550.4123441Uphsa-miR-4677-3p2.65116542.77538440.02470740.4136901Uphsa-miR-708-3p1.58378764.8998950.02688260.4341984Uphsa-miR-518d-5p4.59331271.77068090.02844910.4341984Uphsa-miR-520c-5p4.59329671.77068090.02848860.4341984Uphsa-miR-526a4.59325491.77068090.02859190.4341984Uphsa-miR-16-5p1.181193710.0140530.02954330.4384491Uphsa-miR-873-5p2.73028293.65443560.03059880.4440228Uphsa-miR-218-5p1.6730554.87193180.03272280.4546377Uphsa-miR-542-3p1.7027244.00296470.03874680.4914672Uphsa-miR-501-5p2.64086561.93044820.04660.5261696Uphsa-miR-185-5p–0.99593414.1610130.05117420.5569459No significant changehsa-miR-206–1.3927415.74629870.06334470.6573682No significant changehsa-miR-7113-5p3.80997031.66120460.06525890.6573682No significant changehsa-miR-509-3-5p3.34645992.10642680.06703690.6573682No significant changehsa-miR-548ad-5p1.59152794.23046390.06794660.6573682No significant changehsa-miR-548ae-5p1.59152354.23046390.06844970.6573682No significant changehsa-miR-1070.96472057.85669260.0698410.6573682No significant changehsa-miR-6511b-5p–1.9091833.26213640.07168490.6573682No significant changehsa-miR-6741-3p–2.2649892.29295630.07231310.6573682No significant changehsa-miR-145-3p1.03984946.72405290.07234870.6573682No significant changehsa-miR-204-3p1.64785425.2089780.07284230.6573682No significant changehsa-miR-330-3p–0.8693867.17905010.07312150.6573682No significant changehsa-miR-4762-3p–2.7991182.00769360.07543120.6573682No significant changehsa-miR-589-5p0.86365026.14292560.07737010.6573682No significant changehsa-miR-671-3p–1.0410895.14095410.07906740.6573682No significant changehsa-miR-323b-3p2.22656373.02135460.07929160.6573682No significant changehsa-miR-125a-5p–1.1123567.76783990.07983320.6573682No significant changehsa-miR-141-3p–2.5778722.36672620.08009670.6573682No significant changehsa-miR-3913-5p1.09226744.84461920.08138340.6573682No significant changehsa-miR-378c–1.3333669.24488390.08148840.6573682No significant changehsa-miR-744-5p–0.7978289.55633710.08229310.6573682No significant changehsa-miR-148a-5p–1.1023755.65354380.08422960.6573682No significant changehsa-miR-5193–2.9965551.73410560.0843130.6573682No significant changehsa-miR-1224-5p–1.3338085.02872360.08568650.6573682No significant changehsa-miR-1298-5p2.12718152.45418490.08632590.6573682No significant changehsa-miR-548ay-5p1.4805353.46988450.08859520.6573682No significant changehsa-miR-326–1.4902043.24920.08886160.6573682No significant changehsa-miR-4755-5p–1.9634282.98187390.08897110.6573682No significant changehsa-miR-331-5p1.36015363.33888490.08912020.6573682No significant changehsa-miR-27a-5p0.91835896.58580810.09147640.6573682No significant changehsa-miR-3928-3p–1.2718815.10733890.09309950.6573682No significant changehsa-miR-509-3p3.10730882.47489860.09352760.6573682No significant changehsa-miR-3157-3p1.64744042.53547580.09392050.6573682No significant changehsa-miR-6810-5p–4.8127791.80994670.09414760.6573682No significant changehsa-miR-27a-3p1.01440349.78331950.09462880.6573682No significant changehsa-miR-15b-3p1.87011933.3805780.09688370.6590269No significant changehsa-miR-34a-5p4.16864581.60791170.09770920.6590269No significant changehsa-miR-3130-3p1.50385183.60046640.0994970.6590269No significant changehsa-miR-371a-5p–4.3373541.63136310.09955310.6590269No significant changehsa-miR-1292-5p–1.0331455.80122750.10072790.6590269No significant changehsa-miR-214-3p1.69976022.98037980.10177580.6590269No significant changehsa-miR-1307-3p–0.86251310.957750.10193220.6590269No significant changehsa-miR-12461.068577211.1201330.10603790.6612336No significant changehsa-miR-6747-3p–2.4033271.99047870.10871320.6612336No significant changehsa-miR-6857-3p–2.3131141.84542110.10937970.6612336No significant changehsa-miR-6859-5p–1.2934823.01404160.10991690.6612336No significant changehsa-miR-2277-3p–4.4861881.6844030.11049250.6612336No significant changehsa-miR-1273h-3p–1.0244394.77443060.11251730.6612336No significant changehsa-miR-4489–1.6526562.81154040.11572860.6612336No significant changehsa-miR-2110–0.7633999.5335660.11601440.6612336No significant changehsa-miR-582-3p0.96925916.87174130.11788040.6612336No significant changehsa-miR-4211.13338573.76034650.11796840.6612336No significant changehsa-miR-6808-3p–1.8642982.03410990.1182890.6612336No significant changehsa-miR-6855-5p2.32068252.69201970.11851410.6612336No significant changehsa-miR-30d-3p1.96583832.23801370.11857740.6612336No significant changehsa-miR-548g-3p2.13659631.92773460.12029410.6612336No significant changehsa-miR-4632-5p–4.4384521.66466510.12060230.6612336No significant changehsa-miR-223-5p1.032683810.9945970.12236150.6612336No significant changehsa-miR-148b-3p0.85718429.11758560.12309290.6612336No significant changehsa-miR-511-5p–0.9542877.69622270.12339120.6612336No significant changehsa-miR-3120-5p–1.5695493.1895440.12389890.6612336No significant changehsa-miR-6807-5p–1.684452.76563620.12580880.6612336No significant changehsa-miR-7706–0.8009768.13641830.12598730.6612336No significant changehsa-miR-6877-5p–0.978644.03466950.12817990.6612336No significant changehsa-miR-6867-5p–4.4203871.63993480.1287130.6612336No significant changehsa-miR-654-3p1.47164824.0584580.12927710.6612336No significant changehsa-miR-532-3p1.33452663.05794250.13109350.6612336No significant changehsa-miR-139-5p–0.7639618.79830530.13110840.6612336No significant changehsa-miR-150-5p–0.9517214.57762610.13150270.6612336No significant changehsa-miR-29c-3p1.5905682.93742160.13218380.6612336No significant changehsa-miR-216b-5p2.23739782.2036770.13259360.6612336No significant changehsa-miR-182-5p0.96614188.88975240.13337840.6612336No significant changehsa-miR-422a–1.0784674.62985080.13380460.6612336No significant changehsa-miR-598-5p–2.6666131.57353920.1346770.6612336No significant changehsa-miR-5001-5p–2.5515532.03696420.13704730.6620332No significant changehsa-miR-134-5p–0.9807169.42155160.13734060.6620332No significant changehsa-miR-12842.41454062.19685350.13788130.6620332No significant changehsa-miR-3150a-3p–2.8117731.56347350.13904330.6627395No significant changehsa-miR-363-3p0.70737629.10435110.14135060.6688546No significant changehsa-miR-1228-5p–1.5837185.62672190.15101840.6974406No significant changehsa-miR-451b1.55565342.3416550.15134680.6974406No significant changehsa-miR-4690-3p–1.5853322.31226930.15460860.6974406No significant changehsa-miR-44921.88907492.69085750.15561860.6974406No significant changehsa-miR-22-5p0.71134847.50086180.15644730.6974406No significant changehsa-miR-382-3p2.66187061.88842540.15693760.6974406No significant changehsa-miR-1268a–1.3440743.51730160.1572150.6974406No significant changehsa-miR-130b-5p–1.3498465.53683020.15731530.6974406No significant changehsa-miR-1908-5p–0.8683765.32057330.15750610.6974406No significant changehsa-miR-13233.17542721.74996440.15871520.6974406No significant changehsa-miR-142-5p0.924594310.7469010.15914030.6974406No significant changehsa-miR-202-5p–2.1482192.10916040.16118120.7016757No significant changehsa-miR-3614-5p0.82358815.102390.16282950.7029907No significant changehsa-miR-4734–2.3886632.05939320.16363640.7029907No significant changehsa-miR-500a-3p0.81909936.92276090.16712410.7088167No significant changehsa-miR-6837-3p2.33387041.53371010.16716350.7088167No significant changehsa-miR-1251-5p3.91765011.53416430.17145350.7155546No significant changehsa-miR-1303–1.5555322.65599090.17162890.7155546No significant changehsa-miR-6729-5p–1.5903272.6585340.17256840.7155546No significant changehsa-miR-3164–1.408142.5424410.17479910.7155546No significant changehsa-miR-4686–2.7121781.76712510.17548590.7155546No significant changehsa-miR-199a-5p–0.8083275.1437260.17663250.7155546No significant changehsa-let-7i-5p0.670520313.7323160.17673540.7155546No significant changehsa-miR-98-5p1.00034125.43526180.17751890.7155546No significant changehsa-miR-148a-3p0.943564616.2903350.17888680.7166447No significant changehsa-miR-424-5p1.89402642.21987140.18088030.7189678No significant changehsa-miR-4738-3p–0.9278434.94219830.18166870.7189678No significant changehsa-miR-548av-5p1.80259982.57862820.18452220.7226147No significant changehsa-miR-548k1.80266042.57862820.18509140.7226147No significant changehsa-miR-10b-5p0.986576313.4901420.18694940.7226147No significant changehsa-miR-6894-5p–2.4249551.62339220.18742970.7226147No significant changehsa-miR-502-3p0.79055246.72115640.18812330.7226147No significant changehsa-miR-548d-5p1.2438272.86000410.19065090.728041No significant changehsa-miR-197-5p2.56812161.55695660.1938980.7361359No significant changehsa-miR-3177-3p–0.8276884.38284170.20480540.7701355No significant changehsa-miR-483-3p–1.9744562.22118160.20521220.7701355No significant changehsa-miR-44431.4411882.01313360.2090350.7769258No significant changehsa-miR-450b-5p1.27731843.62551820.20940110.7769258No significant changehsa-miR-3944-5p–1.9172861.66162470.21246540.7809496No significant changehsa-miR-378a-3p–0.69355313.5292090.21345820.7809496No significant changehsa-miR-1268b–1.2537012.8354880.21407350.7809496No significant changehsa-miR-1301-3p–0.6724486.39650390.21531590.7811183No significant changehsa-miR-23b-3p0.83172056.71901260.21667840.7817181No significant changehsa-let-7f-5p0.981304412.7717150.22033570.7827953No significant changehsa-miR-323a-3p1.54215092.9337770.22068170.7827953No significant changehsa-miR-146a-3p–1.4717242.83360030.22298310.7827953No significant changehsa-miR-3615–0.56001711.590870.22315730.7827953No significant changehsa-miR-3653-3p1.78846592.89437710.226650.7827953No significant changehsa-miR-4750-5p2.19884622.28826480.22701040.7827953No significant changehsa-miR-30b-5p–1.1739842.81233670.22811710.7827953No significant changehsa-miR-4685-3p1.70021632.35987040.22841970.7827953No significant changehsa-miR-29a-3p0.62965727.10611010.22857090.7827953No significant changehsa-miR-338-5p0.98310615.15412940.22896460.7827953No significant changehsa-miR-409-5p–2.4550291.86603020.23039490.7835825No significant changehsa-miR-6735-5p–1.1133512.72528620.23586950.7980454No significant changehsa-miR-451a0.69226917.3080790.23857710.8030457No significant changehsa-miR-543–0.7868275.84795380.24408160.8039067No significant changehsa-miR-320a–0.81534217.4506790.2448790.8039067No significant changehsa-miR-9-5p–1.3712763.5878570.24594020.8039067No significant changehsa-miR-4669–1.5951445.86937760.24634950.8039067No significant changehsa-miR-423-5p0.942799214.8185820.2474310.8039067No significant changehsa-miR-505-5p–1.2712113.17836290.24833590.8039067No significant changehsa-miR-433-3p–0.8475413.96455980.24948750.8039067No significant changehsa-miR-378f–0.7110485.40205750.25067520.8039067No significant changehsa-miR-369-5p1.48647542.46927210.25106320.8039067No significant changehsa-miR-2355-3p–1.150723.77394930.25114390.8039067No significant changehsa-miR-203a-3p–1.0094012.95502790.25243440.8040958No significant changehsa-miR-324-5p–1.1598042.78222150.25395810.8050225No significant changehsa-miR-6866-5p1.215842.67713940.25869370.8156615No significant changehsa-miR-146b-5p–0.5062910.0583820.25981260.8156615No significant changehsa-miR-6850-5p–1.3009323.58139080.26217240.8191319No significant changehsa-miR-6805-5p1.6855671.92144730.26607180.8253837No significant changehsa-miR-1288-3p0.71954333.78221210.26981330.8253837No significant changehsa-miR-671-5p–1.325282.76357740.26984850.8253837No significant changehsa-miR-31981.14730642.83459480.27072230.8253837No significant changehsa-let-7g-5p0.684577311.3755670.27323240.8253837No significant changehsa-miR-15a-5p1.19489873.68488850.27407720.8253837No significant changehsa-miR-125b-5p–0.7320387.84266420.27418470.8253837No significant changehsa-miR-1304-3p0.90435262.91665870.27807660.8253837No significant changehsa-miR-1307-5p0.90638195.22430530.27963020.8253837No significant changehsa-miR-1304-5p0.77626744.53056690.28023730.8253837No significant changehsa-let-7a-3p0.93309174.81543460.28309360.8253837No significant changehsa-let-7c-5p0.68757798.81675550.28614860.8253837No significant changehsa-miR-501-3p0.50044238.05750920.28676480.8253837No significant changehsa-miR-45041.03506082.53260260.28725580.8253837No significant changehsa-miR-6740-5p–1.5811231.73439980.28855720.8253837No significant changehsa-miR-548ah-3p–1.3043423.31975060.28907540.8253837No significant changehsa-miR-548ah-5p1.92948961.70389120.28935560.8253837No significant changehsa-miR-483-5p–0.9305617.5649090.29140310.8253837No significant changehsa-miR-660-5p0.70754084.28822360.29142570.8253837No significant changehsa-miR-21-3p0.95423212.41572150.29314740.8253837No significant changehsa-miR-3934-5p–1.2127192.70025450.29332180.8253837No significant changehsa-miR-636–0.9648913.85631120.29356470.8253837No significant changehsa-miR-548o-3p0.6819336.07712910.2939880.8253837No significant changehsa-miR-6847-5p–1.8498931.8003030.2945090.8253837No significant changehsa-miR-548e-3p1.68746982.81395970.29584370.825581No significant changehsa-miR-548aq-3p1.06581742.53636060.29885530.8304362No significant changehsa-miR-548p–1.2827762.57488820.30251630.8347429No significant changehsa-miR-224-5p–0.8157625.42590380.30296180.8347429No significant changehsa-miR-6842-3p–0.540626.41819740.306620.8353266No significant changehsa-miR-6842-5p–1.101073.13502160.30670380.8353266No significant changehsa-miR-664a-5p–0.5116497.56910250.30701130.8353266No significant changehsa-miR-29b-3p0.91357123.38570840.31041690.8410881No significant changehsa-miR-47921.62819131.76705140.31320490.8451357No significant changehsa-miR-374b-5p1.14942072.79769080.31617450.8482237No significant changehsa-miR-548ar-3p2.03686421.72913850.31736620.8482237No significant changehsa-miR-101-3p0.509177712.222810.31904730.8482237No significant changehsa-miR-19b-3p–0.5136617.86911280.31954520.8482237No significant changehsa-miR-138-5p1.7468352.94301730.32225190.8519453No significant changehsa-miR-127-5p1.33999191.93871150.32382210.8526446No significant changehsa-miR-6767-5p–1.2154242.66487080.32608740.8551609No significant changehsa-miR-378g–0.6752234.94050710.32963510.8602968No significant changehsa-miR-1291–1.0524213.06565290.33143690.8602968No significant changehsa-miR-340-5p0.59215067.14479210.33199820.8602968No significant changehsa-miR-27b-3p0.56060849.23279370.33542140.865732No significant changehsa-miR-133a-3p–1.0105862.7950730.33823560.8695584No significant changehsa-miR-3180-3p–0.8465833.85980080.34111220.8701843No significant changehsa-miR-3180–0.8465823.85980080.34114420.8701843No significant changehsa-miR-548f-3p1.06617863.27496080.34815260.8846056No significant changehsa-miR-454-3p1.57228451.8749250.35276710.8870067No significant changehsa-miR-3605-5p0.82218273.58290530.35285110.8870067No significant changehsa-miR-36560.95755666.4010010.35414520.8870067No significant changehsa-miR-200c-3p–0.7122894.396080.35503010.8870067No significant changehsa-miR-4668-5p1.3920881.62407170.35588940.8870067No significant changehsa-miR-125a-3p0.53996495.17150310.35929550.8870821No significant changehsa-miR-44491.59545641.93204230.35945720.8870821No significant changehsa-miR-409-3p–0.5432169.76194620.36136280.8870821No significant changehsa-miR-191-5p–0.43383810.6199170.36161480.8870821No significant changehsa-miR-378b–0.8808432.20214480.3638230.8870821No significant changehsa-miR-7704–0.9841223.39577060.36414250.8870821No significant changehsa-miR-221-5p–1.0027643.49541760.36739520.8870821No significant changehsa-miR-25-5p–0.4931495.60918140.36794070.8870821No significant changehsa-miR-425-3p0.5418835.31046130.36814590.8870821No significant changehsa-miR-4533–1.7605732.12541060.37040920.8888605No significant changehsa-miR-933–0.9591652.33753050.37530030.8888605No significant changehsa-let-7a-5p0.649732212.9758140.37534050.8888605No significant changehsa-miR-4470–0.8925242.81220580.376950.8888605No significant changehsa-miR-2278–1.7666991.87787930.37704780.8888605No significant changehsa-miR-486-5p–0.45793916.6860460.38320730.8888605No significant changehsa-miR-3940-3p–1.4080481.66087320.38328430.8888605No significant changehsa-miR-3187-3p–0.8921372.59216950.38557470.8888605No significant changehsa-miR-44970.73927053.60146120.38708670.8888605No significant changehsa-miR-199b-5p0.98364113.06200570.38806510.8888605No significant changehsa-miR-619-5p0.79148853.69301730.39191010.8888605No significant changehsa-miR-486-3p–0.5271639.29471090.39220850.8888605No significant changehsa-miR-877-5p0.8871422.78355250.39366750.8888605No significant changehsa-miR-26a-5p–0.43852612.0349810.39416450.8888605No significant changehsa-miR-130a-3p0.79318393.4524870.39622620.8888605No significant changehsa-miR-5189-3p–1.8777461.58503350.39793360.8888605No significant changehsa-miR-219a-1-3p0.61665563.92891590.39856450.8888605No significant changehsa-miR-3613-5p1.04868123.43537810.39904070.8888605No significant changehsa-miR-6868-3p1.57841151.832750.39931110.8888605No significant changehsa-miR-43260.50593124.6958750.39969360.8888605No significant changehsa-miR-142-3p–0.5399925.27611150.39995010.8888605No significant changehsa-miR-188-5p–1.7720541.83302910.40037390.8888605No significant changehsa-miR-628-3p–0.8154033.53649410.40201940.8888605No significant changehsa-miR-3163–1.7724861.73363990.40391950.8888605No significant changehsa-miR-6734-5p–0.7852772.78965610.40393290.8888605No significant changehsa-miR-450a-2-3p1.43172921.75965010.4049950.8888605No significant changehsa-miR-181a-2-3p0.50759865.43326870.40736370.8888605No significant changehsa-miR-95-3p0.85445313.15195620.40839020.8888605No significant changehsa-miR-197-3p–0.4494935.82764890.40953430.8888605No significant changehsa-let-7d-3p–0.567878.82932060.4098230.8888605No significant changehsa-miR-1343-3p–0.9465812.51332190.4110810.8888605No significant changehsa-miR-28-5p–0.6213884.6300080.41748680.8997323No significant changehsa-miR-4746-5p–0.4868394.76261530.42122210.9022778No significant changehsa-miR-766-5p–0.7376943.37272110.4220330.9022778No significant changehsa-miR-215-5p–0.4806047.11617660.42495660.9022778No significant changehsa-miR-5584-5p–1.3797731.78465950.4250090.9022778No significant changehsa-let-7e-5p–0.463965.72918920.42678140.9022778No significant changehsa-miR-204-5p1.05626793.14460460.42808740.9022778No significant changehsa-miR-505-3p1.01153242.20079870.43111190.9022778No significant changehsa-miR-3960–0.8203233.02524960.43125140.9022778No significant changehsa-miR-769-5p0.41313476.3285320.4341830.9022778No significant changehsa-miR-574-5p0.51653054.41206080.43461840.9022778No significant changehsa-miR-4745-5p1.10351721.88345070.43517740.9022778No significant changehsa-miR-6826-5p–1.5498231.66799060.43742850.9022778No significant changehsa-miR-6873-3p–1.8542911.84344490.43753160.9022778No significant changehsa-miR-1255a–1.4927191.84635840.43891870.9022778No significant changehsa-miR-196a-5p1.58973581.72112080.43939410.9022778No significant changehsa-miR-541-3p1.65964731.5370370.44331410.9048498No significant changehsa-miR-411-5p–1.0765072.20874150.44351090.9048498No significant changehsa-miR-5010-3p–0.8832362.71594180.44480370.9048498No significant changehsa-miR-627-5p–0.9185253.62527890.44661780.9057188No significant changehsa-miR-1285-3p0.39455647.18308840.45172630.9132424No significant changehsa-miR-190b–0.9639082.6771740.45450070.9144084No significant changehsa-miR-100-5p0.42004139.59219160.45600860.9144084No significant changehsa-miR-361-5p0.5350134.80838030.45650410.9144084No significant changehsa-miR-5187-5p–0.8674992.90178730.45915330.9148355No significant changehsa-miR-147b–0.5893514.46677790.46233710.9148355No significant changehsa-miR-937-3p–0.9743851.93539140.4643010.9148355No significant changehsa-miR-6510-3p1.01742611.76192310.46441560.9148355No significant changehsa-miR-939-5p–0.6972564.09958990.46512440.9148355No significant changehsa-miR-378h–0.7916172.32606910.46587310.9148355No significant changehsa-miR-2171.30214981.90501080.46652410.9148355No significant changehsa-miR-7641–0.4680294.93669520.46901130.9154936No significant changehsa-miR-548av-3p0.471146.2522220.47335180.9154936No significant changehsa-miR-6741-5p0.59492074.17143290.4753080.9154936No significant changehsa-miR-654-5p–0.4645144.84001680.47639950.9154936No significant changehsa-miR-200b-3p0.70911445.01076980.47660570.9154936No significant changehsa-miR-1255b-5p–0.4513045.60569010.47792320.9154936No significant changehsa-miR-491-5p0.93131272.24976780.47894450.9154936No significant changehsa-miR-25-3p0.348973614.278370.47910030.9154936No significant changehsa-miR-941–0.4094368.26144030.4802180.9154936No significant changehsa-miR-12780.74814352.83171470.48087950.9154936No significant changehsa-miR-136-3p1.13335582.0802220.48562820.9218466No significant changehsa-miR-651-5p0.82967412.88578990.48939420.9261983No significant changehsa-miR-2115-5p–0.6541343.85682410.49471840.9261983No significant changehsa-miR-125b-1-3p–0.7100742.58218460.49518710.9261983No significant changehsa-miR-432-5p0.5194595.77323120.49737340.9261983No significant changehsa-miR-370-3p–0.4492918.52314890.49955610.9261983No significant changehsa-miR-30e-3p0.43030926.20127460.50154770.9261983No significant changehsa-miR-4664-3p0.69642192.92129320.50269160.9261983No significant changehsa-miR-199b-3p0.39839557.74128340.50287770.9261983No significant changehsa-miR-199a-3p0.39839577.74128340.50291590.9261983No significant changehsa-miR-3135a0.847272.2651360.5032140.9261983No significant changehsa-miR-200b-5p–1.0187362.43453790.50418330.9261983No significant changehsa-miR-485-5p–0.4525215.31785140.50578170.9261983No significant changehsa-miR-7854-3p0.56683724.12618320.50635950.9261983No significant changehsa-miR-6500-3p1.84579381.57899950.51020980.9267524No significant changehsa-miR-342-5p–0.3711017.97578910.51177930.9267524No significant changehsa-miR-4429–0.4869076.55450030.51282560.9267524No significant changehsa-miR-106b-3p0.29716589.69627240.51355980.9267524No significant changehsa-miR-1468-5p–0.431494.43149040.51388610.9267524No significant changehsa-miR-192-5p–0.38926810.9070.51517790.9267524No significant changehsa-miR-30b-3p–0.7486552.23924320.51997140.9326946No significant changehsa-miR-200a-3p0.60847556.00343140.52255110.9326946No significant changehsa-miR-99b-3p0.44259545.29623110.52420530.9326946No significant changehsa-miR-125b-2-3p–0.4959674.20761810.52525650.9326946No significant changehsa-miR-128-3p–0.41372211.3839020.52623590.9326946No significant changehsa-miR-92b-3p–0.4328075.47865380.5270510.9326946No significant changehsa-miR-143-5p0.62690913.08705240.52976080.9335995No significant changehsa-miR-39110.93204732.74157640.53042180.9335995No significant changehsa-miR-103a-3p0.305439110.3547310.5324890.933819No significant changehsa-miR-30c-2-3p0.84254852.17526550.53544840.933819No significant changehsa-miR-495-3p0.52917275.46755320.53645520.933819No significant changehsa-miR-26a-2-3p1.51988741.60293440.5375990.933819No significant changehsa-miR-126-3p–0.30727312.4131480.53947430.933819No significant changehsa-miR-3682-3p0.81435842.10604390.54073250.933819No significant changehsa-miR-4448–0.8761611.56481660.54325240.933819No significant changehsa-miR-132-5p0.42505664.38400810.54362330.933819No significant changehsa-miR-93-3p–0.5913152.84805140.54454390.933819No significant changehsa-miR-32-3p1.43284081.7613570.54970120.933819No significant changehsa-miR-20a-5p0.37788387.20584720.55027760.933819No significant changehsa-let-7i-3p–1.3222391.87907770.55057620.933819No significant changehsa-miR-329-3p1.00609241.84287530.55172860.933819No significant changehsa-miR-493-5p1.09141512.07293940.55279740.933819No significant changehsa-miR-6878-5p–0.962071.82759460.55472830.933819No significant changehsa-miR-151a-3p–0.2703915.0426130.55475180.933819No significant changehsa-miR-92a-1-5p–0.9316992.28044790.55630170.933819No significant changehsa-miR-46600.95677592.22838560.55760350.933819No significant changehsa-miR-4753-5p0.81886792.33542970.55771730.933819No significant changehsa-miR-340-3p–0.5254084.26399040.56195480.9344595No significant changehsa-miR-34c-5p–0.9097241.66551840.56218010.9344595No significant changehsa-miR-10b-3p0.46651914.88836030.56239290.9344595No significant changehsa-miR-4661-5p0.52124493.07500910.57075370.9441812No significant changehsa-miR-32-5p0.56113413.12758750.57113570.9441812No significant changehsa-miR-452-5p–0.3587466.02193010.57335420.9444801No significant changehsa-miR-548j-3p–0.7538822.61906970.57421830.9444801No significant changehsa-miR-148b-5p–0.6656222.46644560.57565550.9444801No significant changehsa-miR-1180-3p–0.274829.02817030.57748780.9448464No significant changehsa-miR-17-3p–0.9751261.89032050.57877270.9448464No significant changehsa-miR-3127-5p0.58021732.63557280.58319930.9496986No significant changehsa-miR-195-5p0.74328782.50186880.58822750.9555039No significant changehsa-miR-6515-5p0.4809453.63037550.59777440.9686022No significant changehsa-miR-12940.34636514.93233710.60059490.9707635No significant changehsa-miR-6514-5p0.61986552.40062830.60286920.9720336No significant changehsa-miR-7850-5p–1.0244762.14573570.6046710.9725373No significant changehsa-miR-132-3p–0.5232323.06405870.60917890.9773803No significant changehsa-miR-5581-3p–1.1186941.70233920.61087620.9777013No significant changehsa-miR-6786-3p–0.8200011.85401610.61536520.9824779No significant changehsa-miR-24-1-5p–0.5887332.49939690.6172750.9831234No significant changehsa-miR-296-3p0.61981842.70346050.62007190.9851751No significant changehsa-miR-598-3p0.67287582.25668530.62366780.9884833No significant changehsa-miR-4646-5p–0.5408472.29127290.62627760.9902161No significant changehsa-miR-4440–0.5901122.27935660.63890040.998528No significant changehsa-miR-1273 h-5p–0.4239213.7907860.63914110.998528No significant changehsa-miR-576-3p–0.2376878.73406720.64040480.998528No significant changehsa-miR-45160.53847991.82707860.64440290.998528No significant changehsa-miR-28-3p–0.2163849.23620650.64630920.998528No significant changehsa-miR-140-5p0.36915164.34508040.64755970.998528No significant changehsa-miR-4781-3p–0.7666541.96005320.64788830.998528No significant changehsa-miR-320b–0.31876313.643760.64908340.998528No significant changehsa-miR-4726-5p–1.1150351.93218520.65063760.998528No significant changehsa-miR-615-3p0.49955485.29575990.65134570.998528No significant changehsa-miR-44371.12385671.70133770.65213910.998528No significant changehsa-miR-185-3p–0.2755835.40417650.65470880.998528No significant changehsa-miR-1250-5p–0.4602753.12922870.65726770.998528No significant changehsa-miR-532-5p0.202434210.4635050.65728830.998528No significant changehsa-miR-3620-5p0.53274812.17305180.65812880.998528No significant changehsa-miR-6862-5p0.73397341.65785420.65835560.998528No significant changehsa-miR-339-5p0.35993483.54203690.66013040.998528No significant changehsa-miR-193b-3p0.83553951.83482190.66035390.998528No significant changehsa-miR-24-3p0.211912712.4836150.66194220.998528No significant changehsa-miR-361-3p0.19673878.92902520.66303330.998528No significant changehsa-miR-4487–1.1508091.56979530.66367380.998528No significant changehsa-miR-548 am-3p0.51512262.96106650.66725880.998528No significant changehsa-miR-335-3p0.40410493.41631320.66996150.998528No significant changehsa-miR-4840.2296378.23030320.67010360.998528No significant changehsa-miR-320d0.32037038.77208330.67086870.998528No significant changehsa-let-7d-5p0.27110849.26645150.67169540.998528No significant changehsa-miR-196b-5p0.43297912.75833640.67675310.998528No significant changehsa-miR-27b-5p0.5585642.79125710.67888340.998528No significant changehsa-miR-1287-5p0.36756213.72488610.68083770.998528No significant changehsa-miR-92a-3p–0.25264713.8377650.68251840.998528No significant changehsa-miR-1273c–0.5525222.47816650.68296090.998528No significant changehsa-miR-22-3p–0.18138912.4470270.68533610.998528No significant changehsa-miR-766-3p0.56141172.04063060.68555550.998528No significant changehsa-miR-6503-3p–0.3002073.79723930.68607940.998528No significant changehsa-miR-1285-5p0.99502261.64543970.68648920.998528No significant changehsa-miR-181c-3p0.42928272.69015740.68658360.998528No significant changehsa-miR-44670.5934212.53656320.69027731No significant changehsa-miR-642a-3p–0.4754323.2181890.69403721No significant changehsa-miR-3605-3p–0.6591322.47707230.69592271No significant changehsa-miR-3120-3p–0.3252193.14885790.69868961No significant changehsa-miR-1273d–0.3907832.85242220.70065831No significant changehsa-miR-222-3p0.20871158.7331340.70301741No significant changehsa-miR-181b-5p0.22306567.82561680.70937481No significant changehsa-miR-45080.285009710.4247080.71041691No significant changehsa-miR-548u–0.5643021.98144850.71057771No significant changehsa-miR-99b-5p0.19506439.44669710.71232011No significant changehsa-miR-12610.62356881.68847820.71264221No significant changehsa-miR-381-3p–0.2306755.28898090.71600371No significant changehsa-miR-504-5p0.65952511.8402550.71635791No significant changehsa-let-7f-1-3p–0.4560752.67975660.72479341No significant changehsa-miR-18a-3p–0.4437583.33363270.72603521No significant changehsa-miR-6180.41278252.98917590.7269831No significant changehsa-miR-3138–0.749131.82681810.73098191No significant changehsa-miR-328-3p–0.1721316.75136950.73203521No significant changehsa-miR-6806-3p0.37067073.26658150.73294871No significant changehsa-miR-144-3p–0.1901365.92845790.73395611No significant changehsa-miR-12700.5207362.00350590.73600311No significant changehsa-miR-4659b-3p–0.9173161.59456080.7363331No significant changehsa-miR-514a-3p0.66445751.76271730.73646871No significant changehsa-miR-4654–0.6395382.28145970.74282391No significant changehsa-miR-79760.22802994.05529380.74313021No significant changehsa-miR-873-3p0.29947084.22328170.74610151No significant changehsa-miR-424-3p0.17113237.2134380.74663181No significant changehsa-miR-942-3p–0.570662.27594550.75202991No significant changehsa-miR-214-5p0.4314212.53257430.75351771No significant changehsa-miR-610–0.3835322.74638280.753931No significant changehsa-miR-186-5p–0.14693910.2104790.75535951No significant changehsa-miR-629-5p0.169518911.2647910.75701341No significant changehsa-miR-205-5p–0.4554822.21632670.75750521No significant changehsa-miR-1538–0.3778362.47731780.75819751No significant changehsa-miR-4742-3p–0.5744251.96564150.76165091No significant changehsa-miR-152-3p–0.1747237.87559730.76390431No significant changehsa-miR-1260b–0.2319884.02587880.7648171No significant changehsa-miR-191-3p–0.258573.611250.76625291No significant changehsa-miR-12900.17213728.29027430.76720011No significant changehsa-miR-3158-3p–0.15083410.6370150.76726921No significant changehsa-miR-128-1-5p0.50004032.07667890.76826681No significant changehsa-miR-42860.51626471.84431750.77484521No significant changehsa-miR-335-5p0.19355975.83161060.77602721No significant changehsa-miR-210-3p–0.1962575.32674140.77678071No significant changehsa-miR-26b-5p0.17819767.66481120.7790131No significant changehsa-miR-7-5p0.133792311.0934450.78296571No significant changehsa-miR-3691-5p0.3796012.19074580.78578471No significant changehsa-miR-99a-5p–0.15037213.5698670.78886861No significant changehsa-miR-183-5p0.17127028.48130480.79175231No significant changehsa-miR-6821-5p–0.4926811.82365720.79971481No significant changehsa-miR-6750-5p–0.2565612.97910130.80274271No significant changehsa-miR-18a-5p0.53120051.90859350.80392161No significant changehsa-miR-3679-5p–0.500481.96578910.80706121No significant changehsa-miR-345-5p–0.1260437.11035770.80757581No significant changehsa-miR-155-5p0.18976885.32611390.80804941No significant changehsa-miR-659-5p–0.1767874.48250360.80898721No significant changehsa-miR-4772-5p–0.3840252.21723990.81238321No significant changehsa-miR-576-5p–0.3178233.44461280.81606071No significant changehsa-miR-6829-5p0.65732331.68035280.81622841No significant changehsa-miR-5189-5p–0.2647123.69693120.81978241No significant changehsa-miR-4797-3p0.49772171.57064560.8207531No significant changehsa-miR-378d0.15987436.47448160.82218081No significant changehsa-miR-151a-5p–0.189853.77195240.82317261No significant changehsa-miR-320c–0.15948610.9179390.82353171No significant changehsa-miR-106b-5p–0.1385435.33410270.8242241No significant changehsa-miR-379-5p0.2220084.58389070.82613511No significant changehsa-miR-20b-5p–0.138324.86495190.82702141No significant changehsa-miR-1273f0.27242472.5875970.82929211No significant changehsa-let-7b-5p–0.1078413.3251420.82992651No significant changehsa-miR-3140-3p–0.3521472.09327230.83086511No significant changehsa-miR-145-5p0.17944924.35112820.83211141No significant changehsa-miR-548ap-5p0.26004832.0135020.83254451No significant changehsa-miR-181a-3p–0.2107373.05544960.83339321No significant changehsa-miR-378i–0.121088.79994960.83569991No significant changehsa-miR-3199–0.4643731.99238270.83750081No significant changehsa-miR-3127-3p–0.6228971.80054860.8398741No significant changehsa-miR-429–0.3491532.55108790.84538631No significant changehsa-miR-374a-5p0.22645043.089390.84936381No significant changehsa-miR-382-5p0.10952126.94169820.85134191No significant changehsa-miR-6761-5p0.6236791.54953840.8515641No significant changehsa-miR-6875-5p0.52116681.94003190.85157981No significant changehsa-miR-423-3p–0.08161111.8724070.85511711No significant changehsa-miR-60870.58226171.60758620.85540711No significant changehsa-miR-6882-5p0.51914951.57871260.85633171No significant changehsa-miR-3196–0.5497641.56109050.85973651No significant changehsa-miR-96-5p0.20175213.50279270.86046981No significant changehsa-miR-339-3p–0.1011567.38903870.86105931No significant changehsa-miR-30e-5p–0.0801269.47713370.86192781No significant changehsa-miR-23a-5p–0.1522713.54463310.86531281No significant changehsa-miR-46570.29775582.20538270.86551211No significant changehsa-miR-4741–0.3336541.77722740.86783811No significant changehsa-miR-574-3p–0.1379524.59919810.86852751No significant changehsa-miR-493-3p–0.12035.61544890.8692641No significant changehsa-miR-455-5p–0.1574624.05151210.87127331No significant changehsa-miR-4747-5p–0.6615221.75324460.87360841No significant changehsa-miR-31790.67397581.53721340.87393231No significant changehsa-miR-374a-3p0.30153781.71773520.87806221No significant changehsa-miR-378a-5p0.29666642.44214760.87911341No significant changehsa-miR-760–0.0938725.95053120.88034341No significant changehsa-miR-4639-5p–0.4731421.60209460.88136411No significant changehsa-miR-31760.14166032.6330190.88299531No significant changehsa-miR-3124-5p–0.1733893.12592510.88445191No significant changehsa-miR-625-3p–0.1070026.39897750.88452031No significant changehsa-miR-3150b-3p–0.095925.01805380.88502721No significant changehsa-miR-4466–0.2519131.82755560.88572441No significant changehsa-miR-106a-5p0.14048813.92838780.88758241No significant changehsa-miR-24-2-5p–0.0760835.91032130.88842911No significant changehsa-miR-12540.14209863.16687220.8891561No significant changehsa-miR-10a-5p–0.09212711.2620340.88920131No significant changehsa-miR-17-5p–0.0787096.52551910.88968581No significant changehsa-miR-330-5p–0.0828625.91351670.8897761No significant changehsa-miR-3074-5p0.29563171.90121050.89003591No significant changehsa-miR-136-5p0.18043243.12087970.89618271No significant changehsa-miR-324-3p–0.1335593.00588110.89999551No significant changehsa-miR-550a-3-5p0.06422576.57963740.90140261No significant changehsa-miR-26b-3p–0.3079552.00101250.90211221No significant changehsa-miR-4498–0.3345581.74913140.90611821No significant changehsa-miR-331-3p0.27047662.01276790.90779981No significant changehsa-miR-579-5p–0.2454582.63888520.90808541No significant changehsa-miR-195-3p0.10677414.20953030.91043021No significant changehsa-miR-365b-5p0.27076651.95952750.91052691No significant changehsa-miR-3909–0.3078871.97039240.91471021No significant changehsa-miR-146a-5p–0.04981312.1375340.9152021No significant changehsa-miR-3064-5p0.23253212.11061260.91633361No significant changehsa-miR-625-5p0.36274391.89578840.91653561No significant changehsa-miR-499a-5p0.06958464.79891120.91835911No significant changehsa-miR-548q–0.430381.81649820.91986131No significant changehsa-miR-5009-5p–0.0852353.92493930.92116421No significant changehsa-miR-44730.18097761.96477360.92291541No significant changehsa-miR-616-3p–0.1486832.35644930.92511341No significant changehsa-miR-1260a0.12456253.38670940.92682891No significant changehsa-miR-127-3p–0.0573087.70988320.92801091No significant changehsa-miR-10a-3p0.2019662.41650850.92851911No significant changehsa-miR-942-5p–0.0505385.75541590.93040111No significant changehsa-miR-3150a-5p–0.2480332.18404620.93334781No significant changehsa-miR-5010-5p–0.1896892.3508880.93451511No significant changehsa-miR-19a-3p0.06687185.54691390.93496231No significant changehsa-miR-46470.10210533.75807730.93614561No significant changehsa-miR-30c-5p–0.05086.84280240.9376091No significant changehsa-miR-221-3p0.03994298.24074370.93922161No significant changehsa-miR-30d-5p–0.0373412.7887920.94175391No significant changehsa-miR-200a-5p0.21007912.9142630.94178151No significant changehsa-miR-4732-5p–0.0493046.26717870.94291881No significant changehsa-miR-372-3p0.12243693.88806730.94389891No significant changehsa-miR-93-5p–0.037247.5797070.94480461No significant changehsa-miR-550a-5p0.03950626.15471690.94749481No significant changehsa-miR-21-5p–0.03049511.735240.9477891No significant changehsa-miR-342-3p0.04795214.65904610.95501051No significant changehsa-miR-23b-5p–0.0682613.88070560.956381No significant changehsa-miR-3190-3p0.13267342.94808060.95714011No significant changehsa-miR-363-5p0.13313892.99216040.95848561No significant changehsa-miR-874-3p–0.0748643.62639680.95932451No significant changehsa-miR-4755-3p–0.0804793.13702780.96040281No significant changehsa-miR-1-3p–0.0364187.17981750.96321191No significant changehsa-miR-126-5p–0.0653054.00844070.96478031No significant changehsa-miR-652-3p–0.0293455.91222460.96635231No significant changehsa-miR-129-5p–0.0282946.98213230.96753281No significant changehsa-miR-16-2-3p–0.0275238.8119070.96832081No significant changehsa-miR-1306-3p–0.0314425.77171780.9703661No significant changehsa-miR-92b-5p–0.046484.40999620.97245631No significant changehsa-miR-194-5p0.0221137.91644170.97273061No significant changehsa-let-7b-3p0.04378953.48389390.97349181No significant changehsa-miR-130b-3p–0.0258124.15043860.97718691No significant changehsa-miR-584-5p–0.0150479.19855010.97744081No significant changehsa-miR-151b0.02436524.21123580.97873141No significant changehsa-miR-181a-5p–0.01028210.9394460.98200321No significant changehsa-miR-6852-5p0.0188144.59476720.98626411No significant changehsa-miR-425-5p0.00625138.96398750.98960491No significant changehsa-miR-181d-5p–0.0086515.018253611No significant changehsa-miR-516b-5p–0.1026282.819071111No significant changehsa-miR-641–0.0978432.270386811No significant changehsa-miR-1273a–0.3421871.540993611No significant changehsa-miR-548j-5p0.22381241.935619411No significant changehsa-miR-548az-5p0.1321752.232959811No significant changehsa-miR-371b-5p0.34747161.600509711No significant changehsa-miR-6716-3p0.33155851.592339811No significant changehsa-miR-4306–0.3288551.590447911No significant changehsa-miR-6839-5p0.29734051.692811111No significant changehsa-miR-6774-5p–0.2536021.540093811No significant changehsa-miR-6818-5p–0.2347821.88261811No significant changehsa-miR-6781-5p0.22342651.536186811No significant changehsa-miR-4800-5p0.21543582.014541411No significant changehsa-miR-365a-5p0.18276951.777959911No significant changehsa-miR-500b-3p0.16435852.039601811No significant changehsa-miR-5810.16378641.996926211No significant changehsa-miR-6501-5p0.14223511.706718411No significant changehsa-miR-494-3p–0.1318721.77249511No significant changehsa-miR-556-5p–0.130521.738435811No significant changehsa-miR-6780a-5p0.12351971.98260411No significant changehsa-miR-6815-5p–0.0749962.433705111No significant changehsa-miR-50960.07287142.329442311No significant changehsa-miR-5003-3p–0.0705532.037079511No significant changehsa-miR-4724-5p0.06905481.704881911No significant changehsa-miR-431-5p0.06336662.275235511No significant changehsa-miR-664b-5p–0.0622522.053089311No significant changehsa-miR-3122–0.0607051.737748311No significant changehsa-miR-36900.04404393.098556811No significant changehsa-miR-30c-1-3p–0.0379411.868458811No significant changehsa-miR-3675-5p0.03773251.912371111No significant changehsa-miR-6733-5p–0.0242921.594861811No significant changehsa-miR-4433b-3p–0.0148774.828172211No significant changehsa-miR-412-5p–0.0052982.410213511No significant changehsa-miR-29c-5p–0.0011972.704549311No significant changehsa-miR-144-5p–0.0002544.026345111No significant changehsa-miR-375–2.90792810.6212114.83E-083.16E-05Downhsa-miR-885-3p–5.2739852.40018320.00123260.2345272Downhsa-miR-122-5p–2.15982117.226280.00255470.2345272Downhsa-miR-4433b-5p–2.2743284.52276630.00266410.2345272Downhsa-miR-4446-3p–1.7113384.69007480.00704620.2367778Downhsa-miR-885-5p–4.1349222.50153120.00840430.2494538Downhsa-miR-362-5p–4.250442.11849630.01005740.2623126Downhsa-miR-150-3p–1.4622536.40668430.0104930.2623126Downhsa-miR-3173-5p–2.2546042.67844230.01692410.3683817Downhsa-miR-139-3p–1.4297496.62995380.01826710.3847879Downhsa-miR-378e–2.9475713.60081840.02176260.4111132Downhsa-miR-193b-5p–1.5259967.07637640.02203520.4111132Downhsa-miR-4435–2.0385392.53926220.02392510.4123441Downhsa-miR-3191-3p–3.1123642.26003130.03244870.4546377Downhsa-miR-4665-5p–1.7863674.01037890.03519450.4787914Downhsa-miR-1249-3p–4.3666911.63614650.03677610.4900979Downhsa-miR-1299–2.2319418.51187650.03775540.4914672Downhsa-miR-6884-5p–1.815154.26705090.03913670.4914672Downhsa-miR-1247-5p–4.906651.87845320.04075620.5021471Downhsa-miR-4732-3p–1.1080726.60872680.042290.5113961Downhsa-miR-320e–1.9942685.44740590.04551230.5261696Downhsa-miR-146b-3p–1.4064134.61727670.0466920.5261696Downhsa-miR-6511a-5p–3.2617562.13367660.04673480.5261696Downhsa-miR-193a-5p–1.20292611.1551220.04893390.5415905DownFigure 1
Scanning differentially expressed miRNAs. (a) Differentially expressed miRNAs in GSE199942. |LogFC| >1 andP value <0.05 were set to screen. (b) Heatmap of the differentially expressed miRNAs in GSE199942. (c) Venn diagram representing the intersection of up-regulated expressed miRNAs in the 2 data sets (GSE199942 and PMC8702168). (d) Venn diagram representing the intersection of down-regulated expressed miRNAs in the 2 data sets (GSE199942 and PMC8702168).
(a)(b)(c)(d)


## 3.2. Construction of miRNA-mRNA Network

Next, we analyzed the potential targets of the three differentially expressed serum microvesicles miRNAs by miRWalk, and found a total of 593 genes may be co-regulated by at least two of the three miRNAs, of which 192 genes were co-targeted by hsa-miR-15b-5p and hsa-miR-16-5p, 250 genes were co-targeted by hsa-miR-15b-5p and hsa-miR-184, 92 genes were co-targeted by hsa-miR-16-5p and hsa-miR-184, and 59 genes were co-targeted by the three miRNAs (Figures2(a) and 2(b) and Table 2). Given that miRNA negatively regulates the expressions of its targets, we further intersected the 593 prediction genes with the downregulated-expressed genes of PBMCs of IS patients from GSE22255 [26], and obtained 25 genes (Figure 2(c)). The expression of the 25 intersected genes was shown in Figure 2(d). Then, the network containing 3 miRNAs and 25 mRNAs was visualized by the “ggallouvial” package of R software and was shown in Figure 3.Figure 2
Construction of miRNA-mRNA Network. (a) The miRNA-target gene network for differentially expressed miRNAs. (b) Venn diagram showing the overlap among the predicted targets of the differentially expressed miRNAs. (c) Venn diagram showing the overlap between down-regulated expressed genes and predicted targets of the differentially expressed miRNAs. (d) Heat map of the 25 intersected genes.
(a)(b)(c)(d)Table 2
Target genes of the three miRNAs.
Potential targets of hsa-miR-15b-5pPotential targets of hsa-miR-16-5pPotential targets of hsa-miR-184Potential targets of hsa-miR-15b-5p AND hsa-miR-16-5p NOT hsa-miR-184Potential targets of hsa-miR-15b-5p AND hsa-miR-184 NOT hsa-miR-16-5pPotential targets of hsa-miR-16-5p AND hsa-miR-184 NOT hsa-miR-15b-5pPotential targets of hsa-miR-15b-5p AND hsa-miR-16-5p AND 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 3
Sankey diagram of the miRNA-mRNA Network in IS.


## 3.3. Enrichment Analysis of Differentially Expressed Genes

To explore the potential biological functions of the above downregulated-expressed 25 mRNAs, we performed GO annotation and KEGG pathway enrichment analysis. The enriched GO annotation included oxidoreduction coenzyme metabolic process, pyridine-containing compound metabolic process, and nicotinamide nucleotide metabolic process in the biological process (BP) category. Sin3 complex, oligosaccharyltransferase complex, and tetraspanin-enriched microdomain were included in the cellular component (CC) category. NAD+ kinase activity, oxidoreductase activity, and beta-1,3-galactosyltransferase activity were included in the molecular function (MF) category (Figure4(a)). KEGG pathway analysis showed these genes were mainly involved in various types of N-glycan biosynthesis, nicotinate and nicotinamide metabolism, and pentose phosphate pathway (Figure 4(b)).Figure 4
GO (a) and KEGG (b) analysis of the 25 intersected genes.
(a)(b)


## 3.4. PPI Network Construction and Validation of Predictive Feature Biomarkers

We further analyzed the protein-protein interaction (PPI) network of the above 25 intersected genes by Genemania and identified the RB binding protein 4 (RBBP4) had the most interactions in the network (Figure5(a)). Furthermore, we performed the LASSO algorithm to narrow down the above 25 intersected genes and identified eight key genes related to IS (Figure 5(b)). The eight genes were BCL11A, RNLS, UBFD1, STT3A, NADK2, GPR26, PPARA, and SAMHD1. Moreover, we used ROC curve to estimate the predictive value of the eight genes, and found that all of the eight genes had a certain diagnostic value in discriminating IS patients from the healthy controls, with an AUC of 0.7 (Figure 5(C)).Figure 5
PPI network construction and validation of predictive feature biomarkers. (a) PPI network of the 25 intersected genes constructed by GENEMANIA. (b) A plot of biomarkers selection by LASSO regression algorithm. (c) ROC diagnostic curve for the eight genes selected by ASSO regression algorithm.
(a)(b)(c)


## 3.5. Immune Cell Patterns in the Peripheral Bloods of IS

Given that peripheral immune system has been shown to play crucial roles in the evolution of ischemic brain damage [27, 28], we further compared the different immune cell patterns of the peripheral bloods between IS patients and normal controls from GSE22255. As shown in Figures 6(a) and 6(b), M0 macrophages and resting mast cells were significantly decreased in the peripheral bloods of IS patients. M1 macrophages and activated mast cells were slightly increased in the IS group but had no significant difference compared to the normal control group (Figure 6(b)). Furthermore, we analyzed the correlation among the immune cells and found that monocytes were significantly positively correlated to M2 macrophages and significantly negatively correlated to activated NK cells and activated mast cells (Figure 7(a)). Moreover, we analyzed the correlation between the immune cells and the eight variables related to IS and found that STT3A was significantly negative-correlated to follicular helper T cells, activated NK cells and resting dendritic cells (Figure 7(b)).Figure 6
Immune cell patterns in the peripheral bloods of IS samples. (a) The composition of immune cells in the peripheral bloods of IS patients. (b) Comparison of the indicated immune cell subtypes between stroke and normal control group.∗P<0.05.
(a)(b)Figure 7
Correlation analysis of immune cells in the peripheral bloods of IS. (a) The correlation between immune cells in IS samples. (b) The correlation between the eight key genes and immune cells. *P<0.05, **P<0.01, ***P<0.001.
(a)(b)


## 3.6. Verification of the Diagnostic Efficacy of the Key miRNAs

GSE117064 were included in this study as the verification series to validate the diagnostic efficacy of the above three miRNAs. Has-miR-16-5p was more advantageous for diagnosing stroke than the other two miRNAs, as shown in Figure8.Figure 8
Verification of the diagnostic efficacy of the key miRNAs. ROC diagnostic curve for the three miRNAs.


## 4. Discussion

Stroke causes great burden on human health and the economy. Although prominent improvements in the early diagnosis and treatment of IS in the past decade, it is still a leading cause of death and disability. Thus, identifying new biomarkers for the early diagnosis of IS is essential. Recently, serum glutamic oxaloacetic transaminase is reported to may be utilized as predictor in detection of early neurological deterioration in acute IS [29]. Here, we first used the GEO data set to detect the differentially expressed exosomal miRNAs associated with IS and identified three miRNAs (hsa-miR-15b-5p, hsa-miR-184, and hsa-miR-16-5p) intersected with those differentially expressed miRNAs from freshly removed human stroke brain tissue [25]. Among the predicted target genes of the three differentially expressed exosomal miRNAs, 25 mRNAs were obtained by intersecting with differentially expressed genes from GSE22255. Then, we used LASSO algorithm to identify eight key genes related to IS, and further analyzed the independent prediction ROC curve. Subsequently, we analyzed the immune cell patterns in the peripheral bloods of IS patients and the correlation between the immune cells and the eight key genes as potential biomarkers for IS.Previous studies have reported the significance of the miRNAs, which were identified in our study. hsa-miR-15b-5p is highly expressed in the plasma of Alzheimer’s disease (AD) patients, and could distinguish AD patients from normal controls [30, 31]. Besides, hsa-miR-15b-5p in the aqueous humour of patients with diabetic macular oedema shows a fold change greater than –50 in log2 values, compared to the control group [32]. In this study, we firstly identified hsa-miR-15b-5p is highly expressed in the exosomes of IS patients. Recently, hsa-miR-15b-5p is found to regulate the proliferation and apoptosis of human vascular smooth muscle cells [33]. However, the specific mechanism of action of hsa-miR-15b-5p in stroke is unclear, and needs further investigation. miR-16-5p is found to be highly expressed in the in serum and plasma of patients with colorectal cancer [34], and in the feces of patients with precancerous lesions [35]. Besides, hsa-miR-16-5p may be a good candidate for identification of predictive biomarkers of duloxetine response in patients with major depressive disorder who are responsive to duloxetine treatment [36], suggesting that hsa-miR-16-5p may play important roles in the nervous system disease. Consistently, we also found hsa-miR-16-5p is highly expressed in the serum of IS patients, and the specific role of hsa-miR-16-5p in IS needs further investigation. Hsa-miR-184 has been proposed as a biomarker for several cancers, including non-small cell lung, uterine corpus endometrial carcinoma, and oral squamous cell carcinoma [37–39], and brain-enriched hsa-miR-184 is downregulated in older adults with major depressive disorder [40]. Here, we identified that hsa-miR-184 is highly expressed in the serum of IS patients. Moreover, the co-regulated genes of the above three miRNAs were enriched in the N-glycan biosynthesis, nicotinate and nicotinamide metabolism, and pentose phosphate pathways. Interestingly, N-glycosylated immunoglobulin G has been shown to be associated with IS [41], and plasma N-glycans could as emerging biomarkers of cardiometabolic risk [42]. How the three miRNAs regulating N-glycan biosynthesis to affect the IS development would be explored in our following studies, and it could be a very interesting area.Peripheral immune cells interplay with the central nervous system, and play important roles in the brain development, and in the process of IS injury [43, 44]. The immune system is rapidly activated after IS, and peripheral immune cells migrate and infiltrate across the blood–brain barrier into the ischemic region to affect infarction progression and prognosis. Leukocytes are found to aggregate in the ischemic region of MCAO models as early as 30 min after occlusion [45]. Yilmaz et al. fond that CD4+ and CD8+ T cells are distributed in the brain tissue 24 h after IS onset, and act detrimental effects on post-ischemic cerebral immune responses [46]. In this study, we analyzed the different immune cell patterns of the peripheral bloods between IS patients and normal controls from GSE22255, and found M0 macrophages and resting mast cells were significantly decreased in the IS group, and pro-inflammatory M1 macrophages, and activated mast cells were slight increased, suggesting that the inflammatory responses were rapidly activated after stroke. Our result is consistent with recent study that CD11b + CD45+ and CD11b + Ly6G- monocytes and macrophages were greatly increased 3 days after cardiac arrest and resuscitation [47]. Mast cells are rapidly activated and release TNF-α and histamine to aggravate brain damage after IS [48, 49]. Among the eight key genes potentially targeted by the above three miRNAs identified in our study, STT3A, a catalytic subunit of the oligosaccharyltransferase, increases amyloid-β production by promoting N-glycosylation in the pathogenesis of Alzheimer’s disease [50]. Besides, the expression level of STT3A was found to be negatively related to the biomarker expressions of M1 macrophages in breast cancer [51], indicating that STT3A may be involved in the regulating macrophage activation. Although the mechanism of regulating macrophages or mast cells has been extensively study, whether hsa-miR-15b-5p/hsa-miR-184/hsa-miR-16-5p-STT3A axis is involved in the regulating macrophages or mast activation in IS need further investigation.This study has some limitations. For example, our analytical data are derived from public database with relatively small sample sizes. Second, important analysis results need to be further validated with clinical samples. Further large-scale basic studies can be carried out to verify the conclusions of this study.


## 5. Conclusions

We for the first time conducted a comprehensive bioinformatics analysis and identified three serum exosomal miRNAs (hsa-miR-15b-5p, hsa-miR-184, and hsa-miR-16-5p), which may be involved in affecting the peripheral immune cell patterns of IS patients and might serve as promising diagnostic biomarkers. However, the specific pathogenesis and molecular targets still need to be further confirmed through molecular experiments.



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*Source: 9264555-2022-10-10.xml*