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MicroRNA Differential Expression Analysis for Identification of Diagnostic Biomarkers for Canine Visceral Hemangiosarcoma

ABSTRACT: Canine visceral hemangiosarcoma (HSA) is a highly aggressive malignancy of endothelial cells with a poor prognosis and limited diagnostic tools. MicroRNAs (miRNAs) are small non-coding RNAs with emerging utility as diagnostic and prognostic biomarkers due to their stability and disease-specific expression profiles. This study aimed to identify and validate miRNA biomarkers for canine splenic and cardiac HSA using small RNA sequencing and quantitative PCR. Formalin-fixed, paraffin-embedded (FFPE) tissues were analyzed from 24 dogs with histologically confirmed HSA (18 splenic, 6 cardiac) and six non-neoplastic controls (six cardiac and six splenic). A total of 67 and 71 miRNAs were differentially expressed (DE) in splenic and cardiac HSA, respectively, with 18 miRNAs shared between both tumor types. Forty candidate diagnostic miRNAs were selected for validation by RT-qPCR using customized panels. Thirteen miRNAs were validated as DE in each tissue type, showing strong concordance with sequencing results. Pathway enrichment analysis of validated miRNAs revealed significant involvement in oncogenic signaling pathways, including the PI3K-Akt, MAPK, HIF-1, and Ras pathways. These results highlight miRNA signatures with potential diagnostic relevance in visceral HSA and support their utility as biomarkers in archival tissues, with possible future application to liquid biopsy approaches.

ORGANISM(S): Canis lupus familiaris

PROVIDER: GSE310480 | GEO | 2026/04/08

REPOSITORIES: GEO

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Project description:Background: Laryngeal squamous cell carcinoma (LSCC) is the most common type in head and neck squamous cell carcinoma (HNSCC), and the development of LSCC are multistep processes accompanied by changes of molecular biology. Objective: The purpose of this study was to investigate the miRNAs basis of tumorigenesis in LSCC, and analyze the microRNAs dysregulation related with the six targeted genes (CDK1,CDK2, CDK4, MCM2, MCM3, MCM4) which were related to tumorigenesis from the screened miRNAs. Methods: A total number of 5 patients who underwent surgery for primary laryngeal squamous cell carcinoma were recruited for miRNA array analysis. LSCC tissues compared with corresponding adjacent non-neoplastic tissues were analysed by the Affymetrix GeneChip miRNA Array 3.0 to screen effective miRNAs, then mirfocus 3.0 database (http://mirfocus.org/index.php) was adopted to select putative regulated miRNAs of the six targeted genes. Moreover, the selected putative regulated miRNAs were also validated by qRT-PCR in another 36 patients diagnosed for LSCC. Results: Analysed by the miRNAs arrays, there were 127 miRNAs significantly related to tumorigenesis ,and 78 showed a higher expression in tumor than in non-tumor tissue while 49 presented the contrasting pattern(P<0.01).Then analyzed by mirfocus 3.0 database, there were 2 putative regulated miRNAs, hsa-miR-24-3p and hsa-miR-183-5p, corresponding to three of the six targeted genes. Among the 2 putative miRNAs, hsa-miR-24-3p regulated MCM4,CDK1 and CDK4,while hsa-miR-183-5p regulated MCM4 only. Another miRNA we should focus on is hsa-miR-30a-5p, a tumor-suppressive factor, which was down-expressed obviously analysed by the miRNAs arrays. The expression of the 3 putative regulated miRNAs were also validated by qRT-PCR in another 36 patients (P<0.05). Conclusions: The research revealed 127 miRNAs expression signature of tumorigenesis in laryngeal squamous cell carcinoma,and analyzed 3 putative regulated miRNAs, hsa-miR-24-3p, hsa-miR-183-5p and hsa-miR-30a-5p, as potential diagnostic and therapeutic markers in LSCC. The result will contribute to the understanding of the molecular basis of LSCC and help to improve the treatment.

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MicroRNA Differential Expression Analysis for Identification of Diagnostic Biomarkers for Canine Visceral Hemangiosarcoma

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