Directed evolution of genetically encoded LYTACs for cell-mediated delivery

Area of Science:

• Biotechnology
• Molecular Biology
• Therapeutic Drug Development

Background:

• Lysosome-targeting chimeras (LYTACs) offer a novel therapeutic strategy for extracellular protein degradation.
• Existing LYTACs utilize synthetic glycopeptides, limiting their genetic encoding and cellular integration.

Purpose of the Study:

• To design and develop a fully genetically encodable LYTAC (GELYTAC).
• To enable the integration of LYTAC technology into therapeutic cells for targeted delivery and protein degradation.
• To enhance the potency and secretion of GELYTAC from cells.

Main Methods:

• Replaced the glycopeptide component of LYTACs with insulin-like growth factor 2 (IGF2).
• Employed directed evolution to improve GELYTAC efficacy and potency.
• Validated GELYTAC secretion and function in HEK293T cells and primary human T-cells.

Main Results:

• Successfully engineered a genetically encodable LYTAC (GELYTAC) using IGF2.
• Demonstrated enhanced GELYTAC potency through directed evolution.
• Confirmed secretion of functional GELYTAC from both cell lines and primary T-cells, facilitating target protein uptake and degradation.

Conclusions:

• GELYTAC represents a significant advancement in targeted protein degradation technology.
• Genetically encodable LYTACs can be integrated into immune cells for localized therapeutic delivery.
• Engineered immune cells secreting GELYTAC hold promise for spatially selective protein degradation therapies.