Cancer cell killing by target antigen engagement with engineered complementary intracellular antibody single domains...

Area of Science:

• Molecular Biology
• Immunology
• Cancer Research

Background:

• Intracellular tumor proteins are challenging drug targets.
• Previous Antibody-antigen Interaction Dependent Apoptosis (AIDA) approach used anti-β-galactosidase.

Purpose of the Study:

• To develop a new AIDA strategy targeting mutant RAS proteins.
• To induce programmed cell death (PCD) in cancer cells expressing mutant RAS.

Main Methods:

• Co-expressed anti-RAS heavy chain (VH) and light chain (VL) variable regions fused to procaspase-3.
• Engineered intracellular antibodies for specific binding to mutant RAS.
• Utilized a tri-molecular complex to auto-activate caspase pathway.

Main Results:

• Achieved RAS-specific programmed cell death in mutant RAS-expressing cells.
• Demonstrated auto-activation of caspase pathway via tri-molecular complex formation.
• Confirmed requirement of both anti-RAS variable regions for cell death induction.

Conclusions:

• The AIDA strategy is effective for targeting intracellular mutant RAS proteins.
• This method offers a generalizable approach for inducing PCD against various intracellular targets.
• AIDA represents a promising therapeutic strategy for cancers driven by intracellular oncoproteins.