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Profile analysis of circRNAs in human THP-1 derived macrophages infected with intracellular Staphylococcus aureus
Xiaolu Xie1, Zhihao Chen2, Mingxiao Han1
1Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, China.
Background:
Intracellular Staphylococcus aureus (S. aureus) infection is generally persistent, recurrent and difficult to treat due to the poor availability of antibiotics within macrophages cells and the lack of ideal diagnostic markers. Circular RNAs (circRNAs), with covalently closed circular structures, exists in the serum stably and is not easily degraded by nucleases. Besides, circRNAs play a pivotal in the eukaryotic regulation of genes expression and served as biomarkers in variety disease including microbial infections. However, the function of host circRNAs in intracellular S. aureus infection remains largely unclear.
Methods:
In this study, the circRNAs expression profile was investigated by RNA sequencing technology in both S. aureus-infected THP-1 derived macrophages and mock control cells. The differentially expressed circRNAs (DE circRNAs) with a fold-change >1.5 (p < 0.05) are analyzed using functional pathway clustering prediction. Then, RT-qPCR was performed to verify the top 2 up-regulated circRNAs in the THP-1 cell and human serum samples so as to evaluate the value of circRNAs for S. aureus diagnosis.
Results:
An intracellular survival THP-1 derived macrophages model of S. aureus infection was established. A total of 5,299 circRNAs were identified in human THP-1 derived macrophages infected with intracellular S. aureus. There were 61 DE circRNAs with a fold-change >1.5 (p < 0.05) after S. aureus infection. Among them, 22 circRNAs were up-regulated while 39 circRNAs down-regulated. GO and KEGG pathway analysis demonstrated that DE circRNAs were enriched in the processes such as Neurotrophin, Pyruvate metabolism and Notch signaling pathway. Moreover, hsa_circ_0000311 and chr13:43500472-43544806-(novel) were verified to be significantly upregulated in THP-1 derived macrophages and human serum samples between two groups. Finally, the networks of circRNA-miRNA-mRNA based on these two circRNAs were constructed respectively.
Conclusion:
Our study provides the first profile analysis of host circRNAs involved in intracellular S. aureus infection, which may serve as biomarkers for S. aureus diagnosis and contribute to the understanding of S. aureus evasion mechanisms.
Insights
This study identifies host circular RNAs (circRNAs) as potential biomarkers for diagnosing intracellular Staphylococcus aureus infections. These circRNAs may also shed light on how S. aureus evades immune responses.
Area of Science:
- Molecular Biology
- Immunology
- Microbiology
Background:
- Intracellular Staphylococcus aureus (S. aureus) infections are persistent and difficult to treat due to poor antibiotic penetration into macrophages.
- Current diagnostic markers for intracellular S. aureus are lacking.
- Circular RNAs (circRNAs) are stable molecules involved in gene regulation and show potential as biomarkers for various diseases, including infections.
Purpose of the Study:
- To investigate the expression profile of host circRNAs during intracellular S. aureus infection.
- To identify differentially expressed circRNAs (DE circRNAs) and explore their functions.
- To evaluate the diagnostic value of specific circRNAs for S. aureus infection.
Main Methods:
- RNA sequencing was used to analyze circRNA expression in S. aureus-infected THP-1 derived macrophages and control cells.
- Differentially expressed circRNAs (DE circRNAs) were identified using a fold-change >1.5 and p < 0.05 threshold.
- RT-qPCR was performed to validate the expression of top up-regulated circRNAs in cell and human serum samples.
Main Results:
- A total of 5,299 circRNAs were identified, with 61 DE circRNAs found (22 up-regulated, 39 down-regulated).
- Functional analysis indicated DE circRNAs are involved in pathways like Neurotrophin, Pyruvate metabolism, and Notch signaling.
- Two circRNAs, hsa_circ_0000311 and chr13:43500472-43544806-(novel), were significantly upregulated in infected cells and human serum.
Conclusions:
- This study presents the first host circRNA profile analysis in intracellular S. aureus infection.
- Identified circRNAs may serve as novel biomarkers for S. aureus diagnosis.
- The findings contribute to understanding S. aureus evasion mechanisms within host cells.
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