Brain Tumor Res Treat.  2023 Jul;11(3):191-203. 10.14791/btrt.2023.0008.

Comparing the Expression of Canonical and Non-Canonical Inflammasomes Across Glioma Grades: Evaluating Their Potential as an Aggressiveness Marker

• Kim YJ
• Choi W
• Sim J
• Ahn JW
• Park J
• Kim D
• Jeong JY
• Lee JM
• Cho K
• Moon JS
• Moon JH
• Sung KS
• Lim J

• KMID: 2545116
• DOI: http://doi.org/10.14791/btrt.2023.0008

Abstract

Background
Inflammasomes are key in the initiation of inflammatory responses and serve to de-fend the organism. However, when the immune system is imbalanced, these complexes contribute to tumor progression. The purpose of this study was to investigate the effect of non-canonical inflammasomes on glioma malignancy.
Methods
We performed bioinformatics analysis to confirm the expression of canonical and non-canonical inflammasome-related molecules according to the degree of malignancy through immunohistochemical examination of glioma tissues obtained with patient consent from our institution.
Results
Bioinformatics analysis confirmed that the expression levels of non-canonical inflam-masome-related molecules were significantly higher in tumor tissues than in normal tissues, and they also increased according to malignancy, which adversely affected the survival rate. Furthermore, in gliomas, positive correlations were found between N-form gasdermin-D, a key molecule associated with the non-canonical inflammasome, and other related molecules, including NLRP3, caspase-1, caspase-4, and caspase-5. These results were verified by immunohistochemical examination of glioma tissues, and the expression levels of these molecules also increased significantly with increasing grade. In addition, the features of pyroptosis were confirmed.
Conclusion
This study identified the potential of non-canonical inflammasomes as aggressiveness markers for gliomas and presented a perspective for improving glioma treatment.

Keyword

Glioma; Inflammasomes; Disease progression; Biomarkers; Pyroptosis

Figure

• 

  Fig. 1 Analysis of the effects of canonical and non-canonical inflammasome-related gene expression on prognosis and malignancy of glioma patients. A: Non-canonical inflammasome-related genes, including caspase-1, are more highly expressed in glioma tissues than in normal tissues and upregulated according to the malignancy grades, reducing survival time. *p<0.05; **p<0.01; paired t-test.

• 

  Fig. 2 Comparison of canonical NLRP3 inflammasome expression levels according to glioma grade. NLRP3 (red) and caspase-1 (green) were detected in tissues from glioma patients (n=9) through immunohistochemistry. Single and co-expression of molecules for each grade were quantified using ImageJ. *p<0.05; **p<0.01; ***p<0.001; paired t-test with grade II.

• 

  Fig. 3 Comparison of non-canonical inflammasome expression levels according to glioma grade. N-form gasdermin-D (GSDMD-N) (red), caspase-4 (green) (A), and caspase-5 (green) (B) were detected in tissues from glioma patients (n=9) through immunohistochemistry. Single and co-expression of molecules for each grade were quantified using ImageJ. *p<0.05; **p<0.01; ***p<0.001; paired t-test with grade II.

• 

  Fig. 4 Characteristic analysis of inflammatory tumor microenvironment and inflammasome-induced pyroptosis according to glioma grade. A: Glioma tissues of each grade (n=9) were subjected to H&E staining and observed under a Zeiss LSM confocal microscope. B: A TUNEL assay was performed on glioma tissue (n=9), and the frequency of apoptotic cells for each grade was measured using a Zeiss LSM confocal microscope. Representative images are displayed. *p<0.05; ***p<0.001; paired t-test with grade II.

• 

  Fig. 5 Activation and signaling cascades of inflammasomes. Activation of caspase-4/5 (in humans) is involved in the non-canonical inflammasome pathway, whereas activation of caspase-1 is involved in both canonical and non-canonical inflammasome pathways. GSDMD, cleaved by caspase-1/4/5, oligomerizes to form pores in the cell membrane, releasing inflammatory cytokines and triggering pyroptosis. The cleaved GSDMD is involved in the activation of the NLRP3 inflammasome so that the canonical and non-canonical inflammasomes complement each other.

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