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pmid:35313524
Identification of potential biomarkers of vascular calcification using bioinformatics analysis and validation in vivo.
Chuanzhen Chen · Yinteng Wu · Hai-Lin Lu · Kai Liu · Xiao Qin
PeerJ · 2022 doi:10.7717/peerj.13138
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Complete file
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However, we know poorly about the molecular mechanisms and effective therapeutic approaches of VC.\nMethods: The VC dataset, GSE146638, was downloaded from the Gene Expression Omnibus (GEO) database. Using the edgeR package to screen Differentially expressed genes (DEGs). Gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) were used to find pathways affecting VC. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed on the DEGs. Meanwhile, using the String database and Cytoscape software to construct protein-protein interaction (PPI) networks and identify hub genes with the highest module scores. Correlation analysis was performed for hub genes. Receiver operating characteristic (ROC) curves, expression level analysis, GSEA, and subcellular localization were performed for each hub gene. Expression of hub genes in normal and calcified vascular tissues was verified by quantitative reverse transcription PCR (RT-qPCR) and immunohistochemistry (IHC) experiments. The hub gene-related miRNA-mRNA and TF-mRNA networks were constructed and functionally enriched for analysis. Finally, the DGIdb database was utilized to search for alternative drugs targeting VC hub genes.\nResults: By comparing the genes with normal vessels, there were 64 DEGs in mildly calcified vessels and 650 DEGs in severely calcified vessels. Spp1, Sost, Col1a1, Fn1, and Ibsp were central in the progression of the entire VC by the MCODE plug-in. These hub genes are primarily enriched in ossification, extracellular matrix, and ECM-receptor interactions. Expression level results showed that Spp1, Sost, Ibsp, and Fn1 were significantly highly expressed in VC, and Col1a1 was incredibly low. RT-qPCR and IHC validation results were consistent with bioinformatic analysis. We found multiple pathways of hub genes acting in VC and identified 16 targeting drugs.\nConclusions: This study perfected the molecular regulatory mechanism of VC. Our results indicated that Spp1, Sost, Col1a1, Fn1, and Ibsp could be potential novel biomarkers for VC and promising therapeutic targets.", "claims": [{"id": "claim-method", "statement": "Differential expression analysis was performed using t-test to determine P-values and FDR.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:35313524", "locator": "sections[14] \"Statistical analysis\" paragraph 2", "quote": "Differential expression analysis was performed using t-test to determine P-values and FDR."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The raw measurements are available in the Supplemental File.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:35313524", "locator": "sections[32] \"Data Availability\" paragraph 2", "quote": "The raw measurements are available in the Supplemental File."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Comparing the genes with normal vessels, there were 64 DEGs in mildly calcified vessels and 650 DEGs in severely calcified vessels.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:35313524", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "By comparing the genes with normal vessels, there were 64 DEGs in mildly calcified vessels and 650 DEGs in severely calcified vessels."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Comparing the genes with normal vessels, there were 64 DEGs in mildly calcified vessels and 650 DEGs in severely calcified vessels.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:35313524"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. This lane did not download the dataset or read its licence.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE146638"], "outputs": ["the source data with its repository provenance"], "instruments": [], "reagents": []}, {"id": "run_analysis", "order": 3, "name": "Run the published analysis", "kind": "analysis", "state": "complete", "blocked_on": null, "description": "Apply the paper's statistical method to the resolved series and compare the result with the published target.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE146638"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:35313524", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.7717/peerj.13138", "sha256": "sha256:260a2b6a5c03e60ec430894cdc2f53081396f6c5ea95d85113470e0a52a59c84", "bytes": 62143, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "10.7717/peerj.13138", "version": null, "note": "Full article fetched from PMC; 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