Functional repair of p53 mutation in colorectal cancer cells using trans-splicing

Area of Science:

• Molecular Biology
• Cancer Genetics
• Gene Therapy

Background:

• The p53 gene is frequently mutated in human cancers, impacting tumor suppression.
• Current p53-based therapies involve wild-type p53 (wt-p53) administration or mutant p53 suppression.
• Targeting mutant p53 in p53-defective cancer cells remains a therapeutic challenge.

Purpose of the Study:

• To investigate the potential of trans-splicing for correcting mutant p53 transcripts in colorectal cancer (CRC).
• To evaluate the efficacy of trans-splicing in reducing mutant p53 and inducing wt-p53 expression.
• To assess the therapeutic impact of trans-splicing on tumor growth and cancer cell viability.

Main Methods:

• Transfection of human CRC cells with plasmids encoding p53 pre-trans-splicing molecules (PTM).
• Utilizing adenovirus vectors carrying p53 trans-splicing cassettes for intratumoral administration in xenograft models.
• In vitro and in vivo assessment of mutant p53 transcript repair, wt-p53 induction, cell-cycle arrest, and apoptosis.

Main Results:

• p53-PTM plasmids successfully repaired mutant p53 transcripts in p53-mutated CRC cells.
• Trans-splicing led to simultaneous reduction of mutant p53 and induction of wt-p53.
• Adenovirus-mediated trans-splicing suppressed tumor xenograft growth and induced apoptosis in cancer cells.

Conclusions:

• Trans-splicing is a viable strategy for repairing mutant p53 transcripts in colorectal cancer.
• This approach effectively reduces mutant p53, restores wt-p53 function, and inhibits tumor progression.
• Trans-splicing represents a promising new therapeutic avenue for p53-mutated colorectal cancers.