CRISPR/Cas9-Engineered Triple-Fusion Reporter Gene Imaging System for Monitoring Transplanted Neural Progenitor Cells...

Area of Science:

• Regenerative Medicine
• Molecular Imaging
• Neuroscience

Background:

• Neural progenitor cell therapy shows promise for ischemic stroke repair.
• Molecular imaging is vital for assessing cell therapy efficacy.
• The blood-brain barrier hinders effective imaging of transplanted cells.

Purpose of the Study:

• To develop a method for long-term monitoring of transplanted human neural progenitor cells (hNPCs) in an ischemic stroke rat model.
• To combine a CRISPR/Cas9-engineered triple-fusion (TF) reporter gene system with an adenosine agonistic micelle (AM)-based probe delivery strategy.
• To enable noninvasive in vivo tracking of hNPCs.

Main Methods:

• Utilized a CRISPR/Cas9-engineered TF reporter gene system in hNPCs.
• Employed an adenosine agonistic micelle (AM)-based probe delivery strategy.
• Monitored transplanted TF-hNPCs in a rat ischemic stroke model using MRI, bioluminescence imaging, and PET/CT over 8 weeks.

Main Results:

• TF-hNPCs demonstrated proliferation and maturation within the ischemic rat brain.
• Successful in vivo tracking of TF-hNPCs showed migration and differentiation.
• Transplantation of TF-hNPCs improved glucose uptake in the ischemic area and attenuated neurological deficits.

Conclusions:

• A CRISPR/Cas9-engineered TF reporter gene imaging system combined with an AM-based approach enables effective in vivo monitoring of transplanted hNPCs.
• This strategy overcomes blood-brain barrier limitations for evaluating neural progenitor cell transplantation in ischemic stroke.
• The developed system facilitates long-term assessment of cell therapy efficacy in preclinical stroke models.