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MicroRNA Expression Levels Are Altered in the Cerebrospinal Fluid of Patients with Young-Onset Alzheimer’s Disease

McKeever, Paul M., Schneider, Raphael, Taghdiri, Foad, Weichert, Anna, Multani, Namita, Brown, Robert A., Boxer, Adam L., Karydas, Anna, Miller, Bruce, Robertson, Janice, Tartaglia, Maria Carmela·2018·Molecular Neurobiology

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Fig. 1 Workflow for the identification of altered miRNAs in the exosomal cerebrospinal fluid obtained from young-onset Alzheimer’s disease patients versus healthy controls. Pipeline of three phases: CSF sample collection phase, discovery phase, and validation phase. CSF cerebrospinal fluid, YOAD young-onset Alzheimer’s disease, HC healthy controls, LNA locked nucleic acid. Applied Biosystems (ABI) 7900 real-time PCR; receiver operating characteristics (ROC); Gene Set Enrichment Analysis (GSEA) of Gene Ontology Biological Processes terms
Fig. 1 Workflow for the identification of altered miRNAs in the exosomal cerebrospinal fluid obtained from young-onset Alzheimer’s disease patients versus healthy controls. Pipeline of three phases: CSF sample collection phase, discovery phase, and validation phase. CSF cerebrospinal fluid, YOAD young-onset Alzheimer’s disease, HC healthy controls, LNA locked nucleic acid. Applied Biosystems (ABI) 7900 real-time PCR; receiver operating characteristics (ROC); Gene Set Enrichment Analysis (GSEA) of Gene Ontology Biological Processes termsCC BY · PubMed Central
Fig. 2 Discovery phase reveals candidate miRNAs altered in the CSF of YOAD patients versus healthy controls. a Heatmap of differentially expressed miRNAs in the CSF of YOAD patients (n = 17) versus HC (n = 12) identified using one-way ANOVA and pairwise comparisons (p < 0.05). Relative expression level normalized using the geometric mean of spike-ins UniSp6 and cel-miR-39-3p and the endogenous miR-204-5p. Rows are sorted by decreasing fold-change relative to HC. b Six additional candidate microRNAs uncovered in the discovery phase using presence/absence of expression mining and Fisher’s exact test for determining significance. *p < 0.05; **p < 0.01
Fig. 2 Discovery phase reveals candidate miRNAs altered in the CSF of YOAD patients versus healthy controls. a Heatmap of differentially expressed miRNAs in the CSF of YOAD patients (n = 17) versus HC (n = 12) identified using one-way ANOVA and pairwise comparisons (p < 0.05). Relative expression level normalized using the geometric mean of spike-ins UniSp6 and cel-miR-39-3p and the endogenous miR-204-5p. Rows are sorted by decreasing fold-change relative to HC. b Six additional candidate microRNAs uncovered in the discovery phase using presence/absence of expression mining and Fisher’s exact test for determining significance. *p < 0.05; **p < 0.01CC BY · PubMed Central
Fig. 3 Validation phase uncovers miRNAs altered in the CSF of YOAD patients versus healthy controls. Independent validation of discovery phase results with quantitative real-time PCR showing relative expression in YOAD (n = 17) versus HC (n = 12) for a miR-16-5p, b miR-125b-5p, c miR-451a, and d miR-605-5p. Significance between groups was determined using ANOVA followed by pairwise comparisons with Bonferroni correction. *p < 0.05; ****p < 0.0001
Fig. 3 Validation phase uncovers miRNAs altered in the CSF of YOAD patients versus healthy controls. Independent validation of discovery phase results with quantitative real-time PCR showing relative expression in YOAD (n = 17) versus HC (n = 12) for a miR-16-5p, b miR-125b-5p, c miR-451a, and d miR-605-5p. Significance between groups was determined using ANOVA followed by pairwise comparisons with Bonferroni correction. *p < 0.05; ****p < 0.0001CC BY · PubMed Central
Fig. 4 Performance of miRNAs at distinguishing YOAD or LOAD patients from healthy controls. Receiver operating characteristics (ROC) curves plotted as true positive rate (sensitivity) versus false positive rate (100 − specificity) for a YOAD and b LOAD. Area under the curve (AUC) was calculated using the trapezoid method, and the confidence intervals (CI) were calculated using bootstrap sampling in R (v3.4.0)
Fig. 4 Performance of miRNAs at distinguishing YOAD or LOAD patients from healthy controls. Receiver operating characteristics (ROC) curves plotted as true positive rate (sensitivity) versus false positive rate (100 − specificity) for a YOAD and b LOAD. Area under the curve (AUC) was calculated using the trapezoid method, and the confidence intervals (CI) were calculated using bootstrap sampling in R (v3.4.0)CC BY · PubMed Central
Fig. 5 Validated miRNA targets and pathways overlap with sporadic YOAD transcriptome changes. a Five-set Venn diagram showing 2899 mRNAs identified as differentially expressed in the posterior cingulate cortex (PCC) overlap of published microarray data (GSE39420). A total of 874 miRNA targets altered in the exosomes of CSF from sporadic YOAD patients overlapped with the entire microarray dataset. b KEGG pathways were identified using DAVID (v6.8) with the 874 overlapping validated miRNA targets altered in CSF-derived exosomes from sporadic YOAD. Enriched KEGG pathways (y-axis) represented as −log10(p value) (x-axis). The number of genes shared for each pathway is shown at the end of each pathway bar
Fig. 5 Validated miRNA targets and pathways overlap with sporadic YOAD transcriptome changes. a Five-set Venn diagram showing 2899 mRNAs identified as differentially expressed in the posterior cingulate cortex (PCC) overlap of published microarray data (GSE39420). A total of 874 miRNA targets altered in the exosomes of CSF from sporadic YOAD patients overlapped with the entire microarray dataset. b KEGG pathways were identified using DAVID (v6.8) with the 874 overlapping validated miRNA targets altered in CSF-derived exosomes from sporadic YOAD. Enriched KEGG pathways (y-axis) represented as −log10(p value) (x-axis). The number of genes shared for each pathway is shown at the end of each pathway barCC BY · PubMed Central
Fig. 6 Validated miRNA targets share common gene ontologies with transcripts differentially expressed in the posterior cingulate cortex of sporadic YOAD versus healthy controls. Enrichment map representing Gene Ontology (GO) terms for a biological process (BP), b GO molecular function (MF), and c GO cellular compartment enriched in the 883 overlapping targets differentially expressed in the PCC of sporadic YOAD. For the enrichment maps, red nodes = enriched in sporadic YOAD class, blue nodes = depleted in the sporadic YOAD class. Node colors are scaled based on enrichment significance. Encircled node clusters were selected using the Cytoscape (v3.4.0) plug-in AutoAnnotate (v1.1.0). Overrepresented GO terms shown in bold were selected using the WordCloud (v3.1.0) plug-in based on the proportion of redundancy between node clusters. Lists of transcripts shown under enriched GO BP terms are ranked and ≥ 2 fold change (top upregulated transcripts shown next to red nodes and downregulated transcripts next to blue nodes). False discovery rate (FDR) = q < 0.001. Gene sets were pre-ranked using Gene Set Enrichment Analysis (GSEA)
Fig. 6 Validated miRNA targets share common gene ontologies with transcripts differentially expressed in the posterior cingulate cortex of sporadic YOAD versus healthy controls. Enrichment map representing Gene Ontology (GO) terms for a biological process (BP), b GO molecular function (MF), and c GO cellular compartment enriched in the 883 overlapping targets differentially expressed in the PCC of sporadic YOAD. For the enrichment maps, red nodes = enriched in sporadic YOAD class, blue nodes = depleted in the sporadic YOAD class. Node colors are scaled based on enrichment significance. Encircled node clusters were selected using the Cytoscape (v3.4.0) plug-in AutoAnnotate (v1.1.0). Overrepresented GO terms shown in bold were selected using the WordCloud (v3.1.0) plug-in based on the proportion of redundancy between node clusters. Lists of transcripts shown under enriched GO BP terms are ranked and ≥ 2 fold change (top upregulated transcripts shown next to red nodes and downregulated transcripts next to blue nodes). False discovery rate (FDR) = q < 0.001. Gene sets were pre-ranked using Gene Set Enrichment Analysis (GSEA)CC BY · PubMed Central

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Metadata

Authors
McKeever, P. M., Schneider, R., Taghdiri, F., Weichert, A., Multani, N., Brown, R. A., Boxer, A. L., Karydas, A., Miller, B., Robertson, J., & Tartaglia, M. C.
Venue
Molecular Neurobiology
Published
2018-03-30
Volume
55
Issue
12
Pages
8826-8841
ISSN
  • 0893-7648
  • 1559-1182
Cited by
166
References
30
Language
en

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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

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