Cytoskeleton disruption and plasma membrane damage determine methuosis of normal and malignant cells

Area of Science:

• Cell Biology
• Cell Death Mechanisms
• Molecular Biology

Background:

• Methuosis is a cell death characterized by extensive cytoplasmic vacuolization.
• The roles of cytoskeleton and plasma membrane damage in methuosis are not well understood.

Purpose of the Study:

• To investigate the mechanisms of cytoskeleton and plasma membrane damage during methuosis.
• To identify key molecular players involved in methuosis-associated cell damage.

Main Methods:

• Analysis of cytoskeleton protein integrity (F-actin, tubulin, filamin).
• Investigation of RhoA-ROCK1 signaling pathway activation.
• Detection of damage-associated molecular patterns (DAMPs) release.
• Assessment of plasma membrane damage independent of MLKL and GSDMD.
• Evaluation of the role of ESCRT-III components (CHMP3, CHMP5).

Main Results:

• Cytoskeleton proteins (F-actin, α-tubulin, β-tubulin, filamin A/B) were disrupted.
• The RhoA-ROCK1 pathway mediated cytoskeleton disruption.
• Excessive vacuolization led to plasma membrane damage and DAMPs release (LDH, ATP, calreticulin).
• Plasma membrane damage occurred independently of p-MLKL and GSDMD.
• ESCRT-III subunits (CHMP3, CHMP5) negatively regulated vacuolization-induced plasma membrane damage.

Conclusions:

• Cytoskeleton and plasma membrane damage are critical components of methuosis.
• The RhoA-ROCK1 pathway and ESCRT-III complex are key regulators of methuosis.
• Understanding these mechanisms can advance methuosis applications in life sciences and pharmacology.