Engineered extracellular vesicles expressing ICAM-1: A promising targeted delivery system for T cell modifications

Area of Science:

• Biotechnology
• Nanomedicine
• Cell Biology

Background:

• Extracellular vesicles (EVs) are natural nano-carriers with potential for biomolecular delivery.
• Unmodified EVs may exhibit limited accumulation at target sites.
• Engineering EVs for specific cell surface markers can enhance their therapeutic efficacy.

Purpose of the Study:

• To engineer EVs expressing an ICAM-1/LAMP2b fusion protein for targeted delivery.
• To evaluate the uptake and binding of targeted EVs.
• To assess the capacity of engineered EVs to deliver genetic material to T cells.

Main Methods:

• HEK293 cells were modified to produce EVs displaying ICAM-1.
• Targeted and non-targeted EVs were isolated and characterized.
• EVs were loaded with pAAVS1-Puro-GFP plasmids via electroporation.
• Binding affinity and cellular uptake were assessed using ELISA, Western blot, flow cytometry, and imaging.

Main Results:

• Engineered EVs successfully expressed ICAM-1 on their surface.
• Targeted EVs demonstrated significantly higher binding affinity to LFA-1 compared to non-targeted EVs.
• Flow cytometry confirmed differential binding to LFA-1-positive and negative cells.
• EVs efficiently interacted with T cells and delivered encapsulated plasmids.

Conclusions:

• LFA-1-targeted EVs effectively interact with T cells.
• These engineered EVs serve as an ideal delivery system for biomolecules to T cells.
• This approach facilitates advancements in immunotherapies and cell-based treatments.