HSP70 mediates survival in apoptotic cells-Boolean network prediction and experimental validation

Area of Science:

• Computational biology
• Neuroscience
• Cellular signaling

Background:

• Neuronal stress triggers complex survival and apoptosis pathways.
• The precise mechanisms governing cell death and survival signaling remain incompletely understood.
• Heat shock proteins (HSPs) are known to influence cell fate decisions.

Purpose of the Study:

• To elucidate the mechanisms of apoptosis and survival signaling in neuronal cells.
• To construct and validate a Boolean network model of the neuronal apoptotic pathway.
• To investigate the role of HSPs and other proteins in regulating neuronal cell fate.

Main Methods:

• Construction of a Boolean network representing neuronal apoptosis.
• In silico simulations with inputs like FasL and neurotrophic growth factor (NGF).
• Experimental validation using Neuro2a (N2a) cell lines, mRNA level measurements, and protein inhibition experiments.

Main Results:

• Boolean network analysis predicted a survival bias in neuronal cells.
• HSP70 demonstrated a significant pro-survival effect, more so than other HSPs.
• Overexpression of HSP70 reversed FasL-induced apoptosis; inhibition of pro-survival proteins increased apoptosis.

Conclusions:

• Boolean networks provide a reliable tool for analyzing biological networks and predicting cellular fates.
• HSP70 plays a crucial role in neuronal survival by counteracting apoptosis.
• The study validates the pro-survival functions of BCL2, IAP, cFLIP, and NFκB in neuronal cells.