To compare the regulation of BCL-2 effectors BAX and BAK across mammalian species.
Approach:
Cross-species benchmarking: Utilized PBMCs from HCC patients and healthy donors, along with tissues from various mammals, to quantify BAX/BAK localization and OMM residency through selective permeabilization, fractionation, and immunoblotting.
Functional assays: Conducted apoptosis assays in bovine cells and mechanistic follow-up analyses in various cell lines.
Key Findings:
Bovine cells closely recapitulate human-like OMM control of BAX/BAK localization.
Murine systems diverge significantly from human systems.
BAX/BAK localization varies across species, with bovine cells showing a BAX/BAK ratio comparable to humans.
Interpretation:
Species-dependent OMM assembly states of BAX/BAK influence the interpretation of mitochondrial/apoptotic endpoints in drug response assays.
Limitations:
The study focuses on a limited number of species, which may not represent all mammalian variations.
Technical challenges in analyzing primary human PBMCs may affect comparative results.
Conclusion:
Bovine models may provide a more accurate representation of human mitochondrial apoptosis mechanisms, aiding in the selection of preclinical systems.
