Arecoline as a Novel Scaffold Targeting the ATAD2 Bromodomain for Cell Cycle Modulation.

Area of Science:

• Oncology
• Molecular Biology
• Drug Discovery

Background:

• ATPase family AAA domain-containing protein 2 (ATAD2) is an oncogenic chromatin regulator implicated in cancer.
• Arecoline (ARE), a component of areca nut, exhibits context-dependent anti-cancer properties.
• The precise anti-cancer mechanism of arecoline remains to be fully elucidated.

Purpose of the Study:

• To define the anti-cancer mechanism of arecoline.
• To identify direct molecular targets of arecoline in cancer cells.
• To explore the therapeutic potential of arecoline derivatives targeting ATAD2.

Main Methods:

• Breast cancer cell proliferation and colony formation assays.
• Cell cycle analysis and transcriptomic profiling.
• Cellular Thermal Shift Assay-Mass Spectrometry (CETSA-MS) for target identification and validation.
• In silico structure-based design of novel derivatives.

Main Results:

• Arecoline inhibited breast cancer cell proliferation and colony formation by inducing G1/S phase arrest.
• Transcriptomic analysis revealed suppression of the E2F/Cell Cycle gene network.
• CETSA-MS identified ATAD2 as a direct target of arecoline, with ARE binding and stabilizing the protein.
• ARE engagement of ATAD2 led to downregulation of MYC and Cyclin D1, causing G1/S arrest.
• Novel arecoline derivatives with enhanced ATAD2 binding and reduced toxicity were designed.

Conclusions:

• ATAD2 is a druggable target of arecoline in breast cancer.
• Arecoline's scaffold can be repurposed for cancer therapy.
• This study provides a framework for developing safer ATAD2-targeted therapies.