Related Experiment Video
Updated: Jun 18, 2026
Cholinergic Ligand–dependent Modulation of Oxidative Phosphorylation Coupling in Digitonin-permeabilized BE(2)-C Neuroblastoma Cells
Published on: April 28, 2026
BH3 response profiles from neuroblastoma mitochondria predict activity of small molecule Bcl-2 family antagonists
K C Goldsmith1, B J Lestini, M Gross
1The Children's Hospital of Philadelphia, Philadelphia, 19104-4318, USA.
Abstract:
Bcl-2 family proteins regulate mitochondrial apoptosis downstream of diverse stressors. Cancer cells frequently deregulate Bcl-2 proteins leading to chemoresistance. We have optimized a platform for solid tumors in which Bcl-2 family resistance patterns are inferred. Functional mitochondria were isolated from neuroblastoma (NB) cell lines, exposed to distinct BH3-domain peptides, and assayed for cytochrome c release. Such BH3 profiles revealed three patterns of cytochrome c response. A subset had a dominant NoxaBH3 response implying Mcl1 dependence. These cells were more sensitive to small molecules that antagonize Mcl1 (AT-101) than those that antagonize Bcl-2, Bcl-xL and Bcl-w (ABT-737). A second subset had a dominant BikBH3 response, implying a Bcl-xL/-w dependence, and was exquisitely sensitive to ABT-737 (IC(50) <200 nM). Finally, most NB cell lines derived at relapse were relatively resistant to pro-death BH3 peptides and Bcl-2 antagonists. Our findings define heterogeneity for apoptosis resistance in NB, help triage emerging Bcl-2 antagonists for clinical use, and provide a platform for studies to characterize post-therapy resistance mechanisms for NB and other solid tumors.
Insights
This study reveals distinct patterns of apoptosis resistance in neuroblastoma (NB) cell lines, identifying specific Bcl-2 family dependencies. This helps predict sensitivity to targeted cancer therapies and understand chemoresistance.
Area of Science:
- Molecular Biology
- Cancer Research
- Cell Death Pathways
Background:
- Bcl-2 family proteins are key regulators of mitochondrial apoptosis.
- Dysregulation of Bcl-2 proteins contributes to chemoresistance in cancer, particularly neuroblastoma (NB).
Purpose of the Study:
- To develop and optimize a platform for inferring Bcl-2 family resistance patterns in solid tumors, specifically neuroblastoma.
- To characterize heterogeneity in apoptosis resistance in NB cell lines and guide the clinical use of Bcl-2 antagonists.
Main Methods:
- Isolation of functional mitochondria from neuroblastoma cell lines.
- Assay of cytochrome c release upon exposure to distinct BH3-domain peptides to generate BH3 profiles.
- Testing sensitivity to small molecule Bcl-2/Bcl-xL/Bcl-w and Mcl-1 antagonists (ABT-737 and AT-101).
Main Results:
- Three distinct patterns of cytochrome c response were identified.
- A subset of NB cells showed Mcl-1 dependence, sensitive to Mcl-1 antagonists (AT-101).
- Another subset exhibited Bcl-xL/-w dependence, highly sensitive to ABT-737.
- Most NB cell lines from relapsed tumors displayed resistance to BH3 peptides and Bcl-2 antagonists.
Conclusions:
- Apoptosis resistance in neuroblastoma is heterogeneous, defined by specific Bcl-2 family dependencies.
- BH3 profiling can effectively triage emerging Bcl-2 antagonists for clinical application in NB.
- The developed platform facilitates studies on resistance mechanisms in NB and other solid tumors.
