Exofection as a Therapeutic Modality: Restoring P-gp Activity via Trophoblast-Derived EV in Neuroinflammatory...

Area of Science:

• Neuroscience
• Cell Biology
• Biochemistry

Background:

• P-glycoprotein (P-gp) is crucial for clearing toxins and maintaining barrier integrity.
• Reduced P-gp at the blood-brain barrier impairs amyloid-beta clearance and promotes neuroinflammation in Alzheimer's disease.
• Trophoblast-derived extracellular vesicles (CTC-EVs) naturally transfer functional P-gp to other cells (exofection).

Purpose of the Study:

• To investigate if CTC-EV-mediated exofection can restore P-gp function in human brain endothelial cells (hBECs).
• To assess if this restoration enhances amyloid-beta (Aβ) clearance under inflammatory conditions.
• To evaluate the therapeutic potential for Alzheimer's disease.

Main Methods:

• Isolated and characterized CTC-EVs.
• Exposed hBECs to LPS with or without CTC-EVs, assessing P-gp expression and function.
• Evaluated Aβ transport across inflamed hBEC monolayers.
• Administered CTC-EVs to 3xTg-AD mice and P-gp knockout mice to assess in vivo effects.

Main Results:

• CTC-EVs restored LPS-induced P-gp reduction in hBECs and enhanced functional efflux.
• EV treatment improved Aβ transport across inflamed hBECs.
• In vivo, CTC-EVs increased P-gp expression, reduced Aβ burden, and decreased neuroinflammation in AD mice, confirming functional transporter recovery.

Conclusions:

• CTC-EVs are natural carriers of functional P-gp, restoring efflux capacity in compromised endothelial barriers.
• Transcriptomic and network analyses reveal coordinated regulation of transporter, trafficking, and inflammatory pathways.
• This exofection strategy holds promise for enhancing Aβ clearance and mitigating neuroinflammation in Alzheimer's disease.