Establishment of FAP-overexpressing Cells for FAP-targeted Theranostics

Area of Science:

• Biochemistry
• Molecular Biology
• Oncology

Background:

• Fibroblast activation protein (FAP) is a key target in cancer theranostics.
• Accurate controls are essential for validating FAP-targeted agents.
• Existing methods lack specificity, hindering reliable evaluation of FAP theranostics.

Purpose of the Study:

• To establish a pair of FAP-expressing and FAP-negative cell lines for precise theranostic evaluation.
• To create HT1080-hFAP and HT1080-vec cell lines as controls.
• To validate the specificity of FAP-targeted theranostics in vitro and in vivo.

Main Methods:

• Molecular construction of recombinant plasmid pIRES-hFAP for cell line generation.
• Verification of hFAP expression using RT-PCR, Western blotting, and flow cytometry.
• Assessment of FAP's physiological functions and PET imaging in xenograft models.

Main Results:

• Successfully generated HT1080-hFAP (FAP-positive) and HT1080-vec (FAP-negative) cell lines.
• Confirmed hFAP expression and enzymatic activity in HT1080-hFAP cells.
• Demonstrated superior selectivity and uptake of 68Ga-FAPI-04 in HT1080-hFAP tumors via PET imaging.

Conclusions:

• Established a validated pair of cell lines for FAP-targeted theranostics.
• The cell lines facilitate accurate in vitro and in vivo evaluation of FAP agents.
• Enables precise visualization and assessment of FAP-targeted diagnostic and therapeutic strategies.