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Diagnostic value of miR-193a-5p in severe pneumonia and its correlation with prognosis

Journal of Cardiothoracic Surgery volume 20, Article number: 87 (2025) Cite this article

Abstract

Background

Severe pneumonia is a common disease in children, with rapid progression and easy complications of respiratory failure, endangering the lives of children. This study aimed to elucidate the clinical significance of miR-193a-5p in severe pneumonia and to provide a new biomarker for the disease.

Methods

A total of 150 children with severe pneumonia and an equal number of healthy children were selected for analysis. Serum miR-193a-5p levels were detected by RT-qPCR. The correlation of miR-193a-5p with CRP, WBC, neutrophil count, and NLR was assessed by Spearman analysis. The diagnostic and prognostic value of miR-193a-5p in severe pneumonia was analyzed using ROC curves. The relationship between miR-193a-5p and the prognosis of severe pneumonia was evaluated using the Kaplan-Meier curve and a multivariate logistic analysis.

Results

Serum levels of miR-193a-5p were markedly elevated in children with severe pneumonia and exhibited a positive correlation with CRP, WBC, neutrophil count, and NLR. miR-193a-5p could effectively distinguish children with severe pneumonia from healthy children, with an AUC, sensitivity, and specificity of 0.862, 70.67%, and 88.67%, respectively. Serum miR-193a-5p expression was increased in children with poor prognosis and had a predictive value for patient prognosis. High expression of miR-193a-5p was linked to survival in children with severe pneumonia and was a risk factor for adverse prognosis.

Conclusion

Serum levels of miR-193a-5p were markedly elevated in children with severe pneumonia, which may be of significance for the early diagnosis of the disease and prognostic assessment.

Peer Review reports

Background

Pneumonia is a disease that results from the infection and subsequent inflammation of the lungs. This infection may be caused by a variety of bacteria, fungi, or viruses [1]. If the disease is not effectively controlled and worsens, it can develop into severe pneumonia, which may then lead to complications such as sepsis and systemic organ failure [2, 3]. Pneumonia represents one of the most prevalent causes of infectious mortality in children globally [4]. Pediatric pneumonia is typified by a rapid onset and progression. In approximately 7–13% of cases, the disease may progress to severe pneumonia, resulting in organ or systemic damage and endangering the child’s life [5, 6]. Therefore, the timely assessment of the severity of pneumonia is critical to preventing severe pneumonia and determining effective subsequent treatment. It is also an important prerequisite for saving children’s lives [7]. The current diagnosis of severe pneumonia is primarily based on the clinical manifestations observed in children and the presence of infectious indicators. Nevertheless, the efficacy of differential diagnosis and the capacity to evaluate the condition are constrained [8]. In light of these considerations, the search for new biomarkers that can rapidly and accurately reflect the condition and prognosis of children with severe pneumonia has become a clinical priority.

MicroRNAs (miRNAs) constitute a class of small-molecule non-coding RNAs that play a pivotal role in a multitude of physiological responses in vivo, including immune regulation, inflammatory responses, and oxidative stress [9]. Some miRNAs have been identified as being aberrantly expressed in a range of lung diseases, including pneumonia, asthma, and lung cancer [10,11,12]. For example, studies have confirmed the potential of serum-dysregulated miR-10a-3p for the diagnosis and prognostic assessment of severe pneumonia [13]. In a previous study, Hermann and colleagues reported the aberrant expression of miR-193a-5p in patients with community-acquired pneumonia and its potential as a biomarker for this disease [14]. Furthermore, Sun et al. identified miR-193a-5p dysregulation in extracellular vesicles derived from bronchoalveolar lavage fluid in patients with pneumonia [15]. Other studies have demonstrated that miR-193a-5p was also implicated in the progression of lung cancer and chronic obstructive pulmonary disease [16, 17]. It is hypothesized that miR-193a-5p plays a role in the progression of pneumonia in children, which may have clinical significance. However, the role of miR-193a-5p in the progression of pediatric pneumonia remains unclear.

In this study, we investigated the level of serum miR-193a-5p in children with severe pneumonia and its diagnostic and disease assessment value, thereby providing a new direction for the study of pediatric severe pneumonia.

Methods

Study population

One hundred and fifty children with severe pneumonia who were admitted to Anhui Hospital, Children’s Hospital of Fudan University from January 2022 to February 2024 were selected as the severe pneumonia group. Inclusion Criteria: the children meet the relevant diagnostic criteria of childhood pneumonia [18]. Exclusion criteria: [1] congenital diseases, hereditary diseases; [2] the combination of hematologic, neurological, or immune system diseases; [3] the combination of severe cardiac, cerebral, hepatic, and renal insufficiency, or malignant tumors; [4] recent history of major surgery or severe trauma; [5] the combination of infections in other parts of the body. In the same period, 150 healthy children matched for sex and age with the severe pneumonia group were selected as the control group. Informed consent was obtained from the families of all subjects. The study was examined and approved by the medical ethics committee of the hospital.

The prognosis of the children was evaluated and the follow-up period was 28 days [19, 20]. Children who were stabilized or improved after treatment were regarded as having a good prognosis, and those who developed multiple organ failure or died after treatment were regarded as having a poor prognosis.

Serum indicator testing

C-reactive protein (CRP) was detected by AU5800 automated biochemistry analyzer (Beckman Coulter, USA) and D-dimer was analyzed by CS5100 automated coagulation analyzer (Sysmex, Japan), both using immunoturbidimetric method. White blood cells (WBC), platelets, neutrophils, and lymphocytes were assayed by BC-5000 automated blood cell counter (Mindray, Shenzhen, China) using electrical impedance and light scattering methods. The neutrophil-to-lymphocyte ratio (NLR) was the value of neutrophil count/lymphocyte count.

RT-qPCR

A total of 5 mL of venous blood was collected from all subjects and then subjected to centrifugation at 3000 rpm for 15 minutes. The resulting upper layer of serum was then extracted. RNA was extracted using TRIzol reagent (Invitrogen, USA) and reverse transcribed to cDNA using a SuperScript IV VILO Master Mix (Invitrogen). The level of miR-193a-5p was determined using Power SYBR Green Master Mix (Applied Biosystems, USA) and StepOne Real-Time PCR System (Applied Biosystems), with U6 serving as the internal reference. The primers for miR-193a-5p and U6 were as follows: miR-193a-5p (forward: 5’-TGGGTCTTTGCGGGC-3’, reverse: 5’-GAATACCTCGGACCCTGC-3’) and U6 (forward: 5’-CTCGCTTCGGCAGCACA-3’, reverse: 5’-AACGCTTCACGAATTTGCGT-3’). The reaction conditions were as follows: 95 °C for 10 min; 95 °C for 15 s, 60 °C for 1 min, 40 cycles. The miR-193a-5p level was calculated using the 2−ΔΔCt formula.

Statistical methods

SPSS 23.0 software was used for data processing and analysis. Normally distributed measures were expressed as mean ± standard deviation and Student’s t-test was used for comparison between groups. Data that did not conform to normal distribution were expressed as median (interquartile range) and nonparametric tests were performed. The diagnostic and prognostic value of serum miR-193a-5p for severe pneumonia was analyzed by using the ROC curve. Correlations between miR-193a-5p and clinical indicators were assessed by Spearman analysis. Factors affecting the prognosis of severe pneumonia were analyzed by multivariate logistic regression. P < 0.05 was considered statistically different.

Results

Serum miR-193a-5p was upregulated in children with severe pneumonia and correlated with clinical indicators

The levels of CRP, D-dimer, WBC, platelet, neutrophil count, and NLR were found to be markedly elevated in children with severe pneumonia versus the control group, whereas no significant difference was observed in lymphocyte count (Table 1). Serum miR-193a-5p levels were elevated in children with severe pneumonia compared with controls (Fig. 1). The results of correlation analysis revealed that miR-193a-5p was positively correlated with CRP (r = 0.324, P < 0.001), WBC (r = 0.261, P = 0.001), neutrophil count (r = 0.529, P < 0.001), and NLR (r = 0.632, P < 0.001) in children with severe pneumonia. Among them, miR-193a-5p was moderately correlated with NLR and neutrophil count, while weakly correlated with CRP and WBC. (Table 2)

Table 1 Comparison of clinical data between controls and children with severe pneumonia
Full size table
Fig. 1
figure 1

Serum miR-193a-5p expression in children with severe pneumonia and controls. ***P < 0.001

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Table 2 Correlation between serum miR-193a-5p and clinical indicators in children with severe pneumonia
Full size table

Diagnostic value of miR-193a-5p in children with severe pneumonia

The ROC curve demonstrated that miR-193a-5p could effectively differentiate between children with severe pneumonia and controls, with AUC, sensitivity, and specificity of 0.862, 70.67%, and 88.67%, respectively (Fig. 2).

Fig. 2
figure 2

ROC curve of serum miR-193a-5p differentiating children with severe pneumonia from controls

Full size image

Prognostic significance of miR-193a-5p in children with severe pneumonia

Children with severe pneumonia were categorized into the good prognosis group (n = 116) and the poor prognosis group (n = 34) based on the progression of the disease and survival status. The levels of CRP, neutrophil count, and NLR were markedly elevated, while lymphocyte count was notably diminished in children with an unfavorable prognosis (Table 3). Serum miR-193a-5p levels were higher in children with poor prognosis compared to the good prognosis group (Fig. 3A). The AUC of miR-193a-5p for predicting poor prognosis in children was 0.801, and the sensitivity and specificity were 67.65% and 87.07%, respectively (Fig. 3B).

Table 3 Comparison of clinical data between the good prognosis group and the poor prognosis group
Full size table
Fig. 3
figure 3

Expression of serum miR-193a-5p in children with severe pneumonia of different prognosis (A) and its ROC analysis (B). ***P < 0.001

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All children with severe pneumonia were classified into two groups based on the mean level (1.44) of serum miR-193a-5p: high miR-193a-5p group (n = 76) and low miR-193a-5p group (n = 74). Of the subjects, 23 children in the high miR-193a-5p group and 11 patients in the low miR-193a-5p group exhibited a poor prognosis. Kaplan-Meier analysis revealed that the progression-free survival rate was higher in children with low levels of miR-193a-5p (log-rank, P = 0.032, Fig. 4A). Furthermore, multivariate logistic regression demonstrated that miR-193a-5p (OR = 2.507, 95% CI: 1.071–5.864, P = 0.034) and NLR (OR = 2.481, 95% CI: 1.047–5.878, P = 0.039) were factors associated with the adverse prognosis of children with severe pneumonia (Fig. 4B).

Fig. 4
figure 4

Serum miR-193a-5p expression was related to survival in children with severe pneumonia (A) and was a risk factor for poor prognosis (B)

Full size image

Discussion

The occurrence of severe pneumonia can initiate the body’s immune response, stimulate the release of inflammatory mediators in large quantities, and damage organ function [21]. Although clinical indicators related to inflammatory response can reflect the condition of severe pneumonia to a certain extent, their levels are easily affected [22]. Especially for children with severe pneumonia, these indicators are affected by age, immune development, and other factors, which make it difficult to accurately assess the condition of the child [23]. Timely and accurate assessment of the condition and the adoption of methods to prevent its deterioration are of great significance in slowing the progression of severe pneumonia.

It has been demonstrated that miRNAs play a significant role in the pathogenesis of severe pneumonia [24, 25]. It has been reported that the downregulation of miR-1287-5p and miR-150-5p in patients with severe pneumonia may be linked to the pathogenesis of the disease [26, 27]. Hermann et al. discovered a significant difference in the expression of serum miR-193a-5p between patients with pneumonia and healthy populations [14]. In this study, serum miR-193a-5p levels were markedly elevated in children with severe pneumonia in comparison to controls. The inflammatory response is inextricably linked to the development and progression of severe pneumonia [28]. Here, clinical indicators such as CRP, WBC, neutrophil count, and NLR were remarkably higher in children with severe pneumonia than in the control group. Correlation analysis also showed that miR-193a-5p expression correlated with infection-related clinical indicators, including NLR, neutrophil count, CRP, and WBC. Notably, miR-193a-5p was weakly correlated with CRP and WBC. This may be attributed to the fact that the levels of these two indicators varied in children. Previous studies have also shown variable changes in clinical indicators in children with severe pneumonia in different cohorts [29]. Previous studies have revealed that miR-193a-5p is significantly overexpressed in patients infected with SARS-CoV-2 who require mechanical ventilation [30]. These findings suggest that miR-193a-5p may be associated with the occurrence and severity of severe pneumonia.

An increasing number of studies have identified the potential diagnostic value of dysregulated miRNAs in severe pneumonia [31]. For instance, miR-4793-3p and miR-1180-3p were identified as potential biomarkers for differentiating between patients with mild and severe pneumonia [32]. The diagnostic significance of miR-193a-5p has been reported in several diseases, such as breast cancer, post-traumatic stress disorder, and tuberculosis [33,34,35]. Currently, there is an absence of precise indicators for the early diagnosis of children with severe pneumonia, which prevents timely access to effective treatment [7]. It is imperative to develop a concise, rapid, and effective diagnostic method in clinical practice. The present study demonstrated that miR-193a-5p could effectively differentiate between control subjects and children with severe pneumonia, exhibiting high sensitivity and specificity. This indicates the potential of miR-193a-5p as a diagnostic biomarker for severe pneumonia, which may facilitate the clinical diagnosis of the disease.

In addition to respiratory failure, children with severe pneumonia who are not treated in a timely manner may develop severe cardiovascular, neurological, and digestive dysfunction, or even multi-organ failure, which can ultimately result in death [36]. Assessment of severe pneumonia is critical for clinicians to identify and treat children at high risk as early and accurately as possible [37]. While certain clinical indicators have demonstrated potential utility in evaluating the severity of pneumonia and forecasting the prognosis, including procalcitonin, CRP, and NLR, these indicators lack sufficient specificity [29, 38, 39]. Prior research has indicated that miRNAs play a significant role in the progression of pneumonia, with the potential to serve as prognostic biomarkers [31, 40]. For example, the aberrant expression of miR-483-3p was shown to be a significant predictor of clinical outcomes in children with severe pneumonia [41]. A recent study has demonstrated that miR-34a is capable of differentiating between mild and severe cases of mycoplasma pneumonia in children, and is associated with the child’s recovery [42]. Herein, the levels of clinical indicators, including CRP, neutrophil count, lymphocyte count, and NLR, were notably disparate between children with poor prognosis and those with favorable prognosis. Furthermore, miR-193a-5p expression was found to be markedly increased in children with poor prognosis than in those with good prognosis. The ROC curve indicated that miR-193a-5p has the potential to predict the risk of poor prognosis in children with severe pneumonia. Further analysis revealed that high expression of miR-193a-5p was associated with short progression-free survival of patients and was a risk factor for poor prognosis. This may offer a more dependable foundation for clinical prediction of children’s prognosis, which helps physicians to judge the condition and enhance the supervision and treatment of children with severe pneumonia.

This study is also subject to certain limitations. The children with severe pneumonia were drawn primarily from the emergency intensive care unit of our hospital, with a single sample source and fewer cases, which has implications for the generalizability and reliability of the results. Further research is required in the form of multicenter and large-sample studies to evaluate the clinical value of miR-193a-5p in severe pneumonia. The development of miRNAs as biomarkers and therapeutic targets, among others, is still at an early stage. Furthermore, there are significant trials in clinical practice. Although the clinical value of miR-193a-5p in severe pneumonia was discovered through the analysis of clinical data, comprehensive clinical validation is required. This is a prerequisite for the potential application of miR-193a-5p in clinical diagnosis and treatment.

In conclusion, the expression of miR-193a-5p was elevated in patients with severe pneumonia, and its dysregulation may facilitate the diagnosis of severe pneumonia. Furthermore, miR-193a-5p was identified as a promising predictor of prognosis in severe pneumonia, offering insights into the potential for early risk stratification and targeted intervention.

Data availability

No datasets were generated or analysed during the current study.

Abbreviations

CRP:

C-reactive protein

miRNAs:

MicroRNAs

NLR:

Neutrophil-to-lymphocyte ratio

WBC:

White blood cells

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  1. Xiaoji Su and Lijuan Zhu contributed equally to the text.

Authors and Affiliations

  1. Clinical Laboratory, Anhui Hospital, Children’s Hospital of Fudan University, No. 39, Wangjiang East Road, Hefei, Anhui, 230011, China

    Xiaoji Su, Lijuan Zhu, Jiajia Zhuo & Shihai Zhang

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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by LJ Z, SH Z and JJ Z. The first draft of the manuscript was written by XJ S and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Shihai Zhang.

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Su, X., Zhu, L., Zhuo, J. et al. Diagnostic value of miR-193a-5p in severe pneumonia and its correlation with prognosis. J Cardiothorac Surg 20, 87 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s13019-024-03256-y

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Journal of Cardiothoracic Surgery

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