# Comprehensive analysis of mRNA–microRNA–lncRNA expression profiles in post-traumatic elbow heterotopic ossification using RNA sequencing and experimental validation

## TL;DR

This study identifies key genes and microRNAs involved in abnormal bone growth after elbow trauma, offering potential targets for treatment.

## Contribution

First comprehensive profiling of mRNA, microRNA, and lncRNA expression in post-traumatic elbow heterotopic ossification using RNA sequencing.

## Key findings

- Identified 2,138 differentially expressed mRNAs, 40 microRNAs, and 905 lncRNAs in post-traumatic elbow HO tissues.
- Hub genes like MMP9, IL6, and CTSK, along with microRNAs hsa-miR-124-3p and hsa-miR-135b, were highlighted as potential therapeutic targets.
- Key pathways involved include bone mineralization, PI3K–Akt, NF-κB, and TNF signaling.

## Abstract

This study aimed to profile the molecular signatures of post-traumatic elbow heterotopic ossification (HO) to identify key regulators and potential therapeutic targets.

Total RNA from post-traumatic elbow HO tissues (n=4) and normal bone tissues (n=6) was subjected to high-throughput sequencing to identify differentially expressed mRNAs (DEGs), microRNAs (DEMs), and lncRNAs (DELs). Bioinformatics analyses included Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, protein-protein interaction network construction, and transcription factor (TF)-microRNA-mRNA network analysis. The expression trends of four most upregulated and four most downregulated DEGs were validated by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR).

We identified 2,138 DEGs, 40 DEMs, and 905 DELs. DEGs were significantly enriched in biological process “bone mineralization,” cellular component “plasma membrane,” molecular function “integrin binding,” and pathways including PI3K–Akt, NF-κB, JAK–STAT, and TNF signaling pathways. Hub genes with high connectivity included MMP9, IL6, MMP3, CTSK, and BGLAP. Integrated network analysis highlighted the transcription factor JUN and key microRNAs (hsa-miR-124-3p, hsa-miR-548c-3p, and hsa-miR-135b). The qRT-PCR results confirmed the expression trends of selected DEGs.

This study, for the first time, profiled the differentially expressed mRNAs, microRNAs, and lncRNAs in post-traumatic elbow HO using high-throughput RNA sequencing. These findings provide valuable insights into the molecular mechanisms of HO following elbow trauma. The identified hub genes (MMP9, IL6, MMP3, CTSK, and BGLAP), key TF (JUN), and key microRNAs (hsa-miR-124-3p, hsa-miR-548c-3p, and hsa-miR-135b) may serve as potential therapeutic targets for preventing and treating post-traumatic elbow HO.

This study, for the first time, profiled the differentially expressed mRNAs, microRNAs, and lncRNAs in post-traumatic elbow HO using high-throughput RNA sequencing.These findings provide valuable insights into the molecular mechanisms of HO following elbow trauma.The identified hub genes (MMP9, IL6, MMP3, CTSK, and BGLAP), key microRNAs (hsa-miR-124-3p, hsa-miR-548c-3p, and hsa-miR-135b), and key transcription factor (JUN) may serve as potential therapeutic targets for the prevention and treatment of post-traumatic elbow HO.

This study, for the first time, profiled the differentially expressed mRNAs, microRNAs, and lncRNAs in post-traumatic elbow HO using high-throughput RNA sequencing.

These findings provide valuable insights into the molecular mechanisms of HO following elbow trauma.

The identified hub genes (MMP9, IL6, MMP3, CTSK, and BGLAP), key microRNAs (hsa-miR-124-3p, hsa-miR-548c-3p, and hsa-miR-135b), and key transcription factor (JUN) may serve as potential therapeutic targets for the prevention and treatment of post-traumatic elbow HO.