# MicroRNA differential expression analysis in canine visceral hemangiosarcoma formalin-fixed, paraffin-embedded tissues

**Authors:** Laura Machado Ribas, Kerstin Muner, Nelly Elshafie, Elizabeth Rozanski, Francisco O. Conrado, Luis Dos Santos, Andrea Pires dos Santos

PMC · DOI: 10.3389/fvets.2026.1755166 · 2026-03-17

## TL;DR

This study identifies microRNA biomarkers in canine hemangiosarcoma tissues that could help diagnose this aggressive cancer using archived samples.

## Contribution

The study identifies and validates miRNA biomarkers specific to canine visceral hemangiosarcoma using FFPE tissues.

## Key findings

- 67 and 71 miRNAs were differentially expressed in splenic and cardiac HSA, respectively.
- 13 miRNAs were validated as differentially expressed in each tissue type with strong concordance.
- Validated miRNAs were linked to key oncogenic pathways like PI3K-Akt and MAPK.

## 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 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, six cardiac) and 12 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 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 a significant involvement in oncogenic signaling pathways, including the PI3K-Akt, MAPK, HIF-1, and Ras pathways.

These results highlight miRNA signatures that may have diagnostic value in visceral HSA and support their use as biomarkers in archived tissues, with potential future application in liquid biopsy approaches.

## Linked entities

- **Diseases:** hemangiosarcoma (MONDO:0016982)
- **Species:** Canis lupus familiaris (taxon 9615)

## Full-text entities

- **Diseases:** malignancy (MESH:D009369)
- **Chemicals:** paraffin (MESH:D010232), Formalin (MESH:D005557)
- **Species:** Hysterothylacium sp. SA (species) [taxon 1884613], Canis lupus familiaris (dog, subspecies) [taxon 9615]

## Figures

3 figures with captions in the complete paper: https://tomesphere.com/paper/PMC13037417/full.md

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Source: https://tomesphere.com/paper/PMC13037417