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Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation

  • Authors:
    • Yiheng Yang
    • Jiahao Zou
    • Peng Yang
    • Zhenzhong Zheng
    • Qingshan Tian
  • View Affiliations / Copyright

    Affiliations: Department of Cardiology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
    Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 56
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    Published online on: December 16, 2025
       https://doi.org/10.3892/etm.2025.13050
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Abstract

The mechanisms underlying the pathogenesis of septic cardiomyopathy (SCM) are intricate and incompletely understood. PANoptosis is a novel type of programmed cell death, and in the present study, bioinformatics, machine learning and experimental validation were used to identify key PANoptosis‑related genes (PRGs) associated with SCM. Differentially expressed genes were obtained through analysis of the Gene Expression Omnibus dataset, and these genes were intersected with the PRGs to obtain the differentially expressed PRGs. Three machine learning algorithms were used to screen key PRGs; CIBERSORT was used for immune infiltration analysis and the diagnostic value of key PRGs was evaluated by plotting receiver operating characteristic curves. Additionally, a competitive endogenous (ce)RNA regulatory network analysis was conducted, and drug prediction analysis was performed. Finally, the expression of key PRGs was verified via quantitative PCR. A total of 157 differentially expressed genes and 21 differentially expressed PRGs were screened. In addition, two key PRGs (RIPK2 and GADD45B) were screened using least absolute shrinkage and selection operator regression, the support vector machine‑recursive feature elimination algorithm and the random forest algorithm, with both genes demonstrating a high diagnostic value. RIPK2 and GADD45B were positively correlated with neutrophils. The ceRNA regulatory network included two mRNAs, eight microRNAs and 16 long noncoding RNAs and 10 drugs/molecular compounds were predicted. Finally, quantitative PCR results revealed that the expression of both RIPK2 and GADD45B was upregulated in the lipopolysaccharide‑induced HL‑1 cell injury model. In conclusion, the present study identified two key PRGs (RIPK2 and GADD45B) associated with SCM; these findings may lead to the development of novel diagnostic and therapeutic approaches for SCM.
View Figures

Figure 1

Flow chart of the present study.
PRGs, PANoptosis-related genes; GO, Gene Ontology; KEGG, Kyoto
Encyclopedia of Genes and Genomes; LASSO, least absolute shrinkage
and selection operator; SVM-RFE, support vector machine-recursive
feature elimination; ceRNA, competitive endogenous RNA; GSEA, gene
set enrichment analysis; ROC, receiver operating
characteristic.

Figure 2

Screening of DEGs. (A) Heatmap of
DEGs in GSE79962, the expression of genes from low to high is shown
in blue to red. (B) Volcano plot of DEGs in GSE79962. Green dots
represent significantly down-regulated genes. Red dots represent
significantly up-regulated genes. Gray dots represent genes with no
significant differential expression. DEGs, differentially expressed
genes; FC, fold change.

Figure 3

Identification of differentially
expressed PRGs and GO/KEGG enrichment analysis. (A) Venn diagrams
were used to show genes where DEGs intersected with PRGs. (B)
Differentially expressed PRGs were used for GO analysis. The top
five functional terms in the categories biological process,
cellular component and molecular function were visualized using
bubble plots. Larger bubbles indicate a higher number of enriched
genes. (C) The top 30 KEGG pathways of differentially expressed
PRGs were shown using bar plots. A redder color indicates a higher
enrichment significance. PRGs, PANoptosis-related genes; GO, Gene
Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; DEGs,
differentially expressed genes.

Figure 4

Identification of key genes by three
machine learning algorithms and evaluation of the diagnostic value
of key genes. (A) Nine differentially expressed PRGs were obtained
using LASSO regression analysis. (B) Four differentially expressed
PRGs were obtained using the RF algorithm (indicated by the red
box). (C) Four differentially expressed PRGs were obtained using
the SVM-RFE algorithm. (D) The two key genes (RIPK2 and GADD45B)
were obtained by intersecting the results of LASSO regression, RF
and SVM-RFE. The Venn diagram showed the results of the
intersection. (E and F) Receiver operating characteristic curves
were used to assess the diagnostic efficacy of key genes (E)
GADD45B and (F) RIPK2. PRGs, PANoptosis-related genes; LASSO, least
absolute shrinkage and selection operator; RF, random forest;
SVM-RFE, support vector machine-recursive feature elimination; AUC,
area under the curve; CI, confidence interval; RMSE, root mean
square error.

Figure 5

External dataset validation of key
gene expression. (A) Comparison of key gene expression between the
control group and the SCM group in the GSE79962 dataset. (B) In the
validation set (combined from GSE53007 and GSE142615), the
comparison of the expression of key genes between the control group
and the SCM group. The above results were presented in box plots.
***P<0.001. SCM, septic cardiomyopathy.

Figure 6

Assessment of immune cell
infiltration and correlation of key genes with immune cells. (A) A
bar-plot diagram was used to show the percentage of 22 immune cell
types in each sample. Each color represents the corresponding type
of cell. (B) A lollipop plot was used to show the correlation
between GADD45B and the 22 immune cell types. (C) A lollipop plot
was used to show the correlation between RIPK2 and the 22 immune
cell types. The size of the dots reflects the strength of the
correlation. Con, control; SCM, septic cardiomyopathy; abs(cor),
absolute value of correlation coefficient.

Figure 7

GSEA analysis of key genes. (A and B)
GSEA analysis of (A) GADD45B-up and (B) GADD45B-down. (C and D)
GSEA analysis of (C) RIPK2-up and (D) RIPK2-down. Enrichment plots
were used to display the five most significantly upregulated or
downregulated pathways. GSEA, gene set enrichment analysis.

Figure 8

Key gene-based ceRNA regulatory
networks and potential therapeutic agents. (A) Construction of
ceRNA networks based on key genes. The ceRNA network consists of
two mRNAs, eight miRs and 16 long non-coding RNAs. (B) Potential
drug prediction based on key genes. 10 drugs or molecular compounds
related to RIPK2 were obtained from the Drug-Gene Interaction
Database. ceRNA, competitive endogenous RNA. miR, microRNA.

Figure 9

Reverse transcription quantitative
PCR to verify the expression of key PRGs. (A) Comparison of GADD45B
expression in the control and LPS-induced HL-1 cell injury group.
(B) Comparison of RIPK2 expression in the control and LPS-induced
HL-1 cell injury group. ***P<0.001. PRGs,
PANoptosis-related genes; LPS, lipopolysaccharide.
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Copy and paste a formatted citation
Spandidos Publications style
Yang Y, Zou J, Yang P, Zheng Z and Tian Q: Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation. Exp Ther Med 31: 56, 2026.
APA
Yang, Y., Zou, J., Yang, P., Zheng, Z., & Tian, Q. (2026). Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation. Experimental and Therapeutic Medicine, 31, 56. https://doi.org/10.3892/etm.2025.13050
MLA
Yang, Y., Zou, J., Yang, P., Zheng, Z., Tian, Q."Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation". Experimental and Therapeutic Medicine 31.2 (2026): 56.
Chicago
Yang, Y., Zou, J., Yang, P., Zheng, Z., Tian, Q."Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation". Experimental and Therapeutic Medicine 31, no. 2 (2026): 56. https://doi.org/10.3892/etm.2025.13050
Copy and paste a formatted citation
x
Spandidos Publications style
Yang Y, Zou J, Yang P, Zheng Z and Tian Q: Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation. Exp Ther Med 31: 56, 2026.
APA
Yang, Y., Zou, J., Yang, P., Zheng, Z., & Tian, Q. (2026). Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation. Experimental and Therapeutic Medicine, 31, 56. https://doi.org/10.3892/etm.2025.13050
MLA
Yang, Y., Zou, J., Yang, P., Zheng, Z., Tian, Q."Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation". Experimental and Therapeutic Medicine 31.2 (2026): 56.
Chicago
Yang, Y., Zou, J., Yang, P., Zheng, Z., Tian, Q."Analysis of PANoptosis‑related genes in septic cardiomyopathy by bioinformatics, machine learning and experimental validation". Experimental and Therapeutic Medicine 31, no. 2 (2026): 56. https://doi.org/10.3892/etm.2025.13050
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