Parallel Automated Flow Synthesis of Covalent Protein Complexes That Can Inhibit MYC-Driven Transcription

Area of Science:

• Molecular Biology
• Cancer Research
• Protein Engineering

Background:

• MYC transcription factor dysregulation is implicated in numerous human cancers.
• MYC/MAX and MAX/MAX dimers regulate gene transcription via enhancer box DNA binding.
• Targeting these MYC-related transcription complexes presents a significant challenge in cancer therapy.

Purpose of the Study:

• To engineer synthetic homo- and heterodimeric protein complexes that mimic the inhibitory MAX/MAX dimer.
• To develop novel protein complex mimetics for attenuating oncogenic MYC-driven transcription.
• To explore the potential of automated flow synthesis for rapid production of these therapeutic agents.

Main Methods:

• Engineered covalently linked synthetic homo- and heterodimeric protein complexes.
• Utilized parallel automated flow synthesis for rapid preparation of protein complexes (∼20 kDa, 167-231 residues).
• Assessed DNA binding activity, cell penetration, and anti-proliferative effects in cancer cell lines. Performed RNA sequencing and gene set enrichment analysis.

Main Results:

• The synthetic covalent dimers demonstrated DNA binding activity and were intrinsically cell-penetrant.
• These engineered dimers effectively inhibited cancer cell proliferation across different cell lines.
• RNA sequencing confirmed interference with MYC-driven transcription in A549 cancer cells.

Conclusions:

• Automated flow technology can rapidly produce engineered synthetic protein complex mimetics.
• These synthetic dimers represent a promising starting point for developing novel inhibitors of MYC-driven cancer growth.
• The study highlights a new strategy for targeting transcription factor dysregulation in cancer.