Related Experiment Video
Updated: Sep 14, 2025
Author Spotlight: Flow Cytometric Determination of Pyroptosis in Avian Cells
Published on: May 31, 2024
Discovery of pyroptosis-inducing natural products in neuroblastomas: computational studies with experimental
Beni Lestari1,2, Rohmad Yudi Utomo3,4, Faaza Aulia Rahman2
1Department Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia.
Background:
Neuroblastomas evade apoptosis due to oncogene mutations and antiapoptotic proteins, necessitating novel therapeutics that work in concert with other forms of cell death. Pyroptosis has potential as a strategic cell death mechanism in neuroblastoma. This study aimed to identify compounds that modulate pyroptosis, specifically those that target gasdermin D (GSDMD) oligomerization.
Methods:
The study employed computational analysis and in vitro screening. The COCONUT database provides the natural compound dataset. Drug-likeness analysis and pharmacophore fitting were applied to screen potential GSDMD oligomerization modulators. Hit compounds underwent molecular docking with MOE and molecular dynamic (MD) simulations with NAMD 2.14 to analyse structural changes. The computational screening results were corroborated by in vitro assays, including the WST-8 assay, Western blot, and immunofluorescence, which target pyroptosis-specific caspase-1.
Results:
Pharmacophore fitting, molecular docking, and molecular dynamics simulations identified the top five compounds, namely, quercetin, naringenin, hesperetin, curcumin, and galangin, as potent modulators of GSDMD oligomerization. Among these compounds, curcumin, quercetin, and galangin exerted potent cytotoxic effects on GSDMD-expressing neuroblastoma SK-N-AS cells, with IC50 values of 21, 37, and 49 µM, respectively. Curcumin and quercetin also promoted apoptosis via increased caspase-3 cleavage and reduced procaspase-7 and -8 levels, as shown by immunoblotting. Curcumin and galangin upregulated caspase-1 expression, as demonstrated by the detection of a fluorescent-labelled inhibitor of caspase-1 by immunostaining, suggesting that these two compounds could induce pyroptosis. Hesperetin and naringenin showed low cytotoxicity, had no effect on caspase activation, and did not exhibit signs of pyroptosis in SK-NA-S cells.
Conclusions:
Our study successfully identified curcumin as a strong regulator of both apoptosis and pyroptosis, quercetin as a strong modulator of apoptosis, and galangin as a strong modulator of pyroptosis. Further research on these compounds is crucial for the development of novel therapeutic strategies for neuroblastoma treatment.
Insights
This study identified natural compounds like curcumin and quercetin that can trigger programmed cell death (apoptosis and pyroptosis) in neuroblastoma cells. These findings offer potential new therapeutic avenues for treating this aggressive cancer.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Neuroblastoma often evades apoptosis due to genetic mutations, necessitating novel therapeutic strategies.
- Pyroptosis presents a promising cell death mechanism for neuroblastoma treatment.
- Targeting gasdermin D (GSDMD) oligomerization is a key strategy to modulate pyroptosis.
Purpose of the Study:
- To identify natural compounds that modulate pyroptosis by targeting GSDMD oligomerization.
- To evaluate the cytotoxic effects and cell death mechanisms of identified compounds in neuroblastoma cells.
Main Methods:
- Computational screening including pharmacophore fitting, molecular docking, and molecular dynamics simulations.
- In vitro validation using WST-8 assays, Western blot, and immunofluorescence targeting caspase-1.
- Drug-likeness analysis of natural compounds from the COCONUT database.
Main Results:
- Five potent GSDMD oligomerization modulators were identified: quercetin, naringenin, hesperetin, curcumin, and galangin.
- Curcumin, quercetin, and galangin exhibited significant cytotoxicity against GSDMD-expressing neuroblastoma cells.
- Curcumin and galangin induced pyroptosis, while curcumin and quercetin also promoted apoptosis.
Conclusions:
- Curcumin strongly regulates both apoptosis and pyroptosis.
- Quercetin is a potent modulator of apoptosis, and galangin is a potent modulator of pyroptosis.
- These compounds represent promising candidates for developing novel neuroblastoma therapeutics.
More Related Videos
06:12Author Spotlight: THP-1 Macrophage Response to LPS/ATP — Unveiling the Pyroptosis, Apoptosis, and Necroptosis Spectrum
Published on: May 3, 2024
07:10Orthotopic Xenografting of Human Luciferase-Tagged Malignant Peripheral Nerve Sheath Tumor Cells for in vivo Testing of Candidate Therapeutic Agents
Published on: March 7, 2011