Aloperine Suppresses Cancer Progression by Interacting with VPS4A to Inhibit Autophagosome-lysosome Fusion in NSCLC

Area of Science:

• Pharmacology
• Oncology
• Cell Biology

Background:

• Aloperine (ALO), a natural alkaloid, shows potential antitumor activity.
• Its precise mechanism of action and molecular targets are not fully understood.

Purpose of the Study:

• To elucidate the mechanism of action and identify the molecular target of Aloperine in non-small cell lung cancer.
• To evaluate Aloperine's efficacy in preclinical cancer models and its potential as an adjunct therapy.

Main Methods:

• In vitro studies on non-small cell lung cancer cell lines.
• In vivo studies using mouse tumor models.
• Autophagy inhibition assays, reactive oxygen species (ROS) detection, and apoptosis assays.
• Identification of Aloperine's direct target using genetic approaches (knockdown, knockout) and binding site analysis.
• Combination therapy studies with anti-PD-L1/TGF-β bispecific antibody.

Main Results:

• Aloperine inhibited proliferation, migration, and tumor development in vitro and in vivo.
• ALO blocked autophagosome-lysosome fusion and autophagic flux, increasing sequestosome-1 (SQSTM1) and ROS levels, inducing apoptosis.
• VPS4A was identified as a direct target of Aloperine, with specific binding sites.
• VPS4A knockout mimicked Aloperine's effects.
• Aloperine enhanced the efficacy of an anti-PD-L1/TGF-β bispecific antibody.

Conclusions:

• Aloperine is a novel late-stage autophagy inhibitor targeting VPS4A.
• It induces tumor cell death via SQSTM1 accumulation, ROS production, and apoptosis.
• Aloperine shows therapeutic potential for non-small cell lung cancer and can enhance immunotherapy.