Method Profile
Use patient- and PDX-derived organoids to compare CPT1A transcript effects and an ASO targeting a THRAP3-binding site.
Mechanistic assessment of a transcript-selective intervention in treatment-naive and resistant prostate cancer models.
Organoids from fresh treatment-naive prostatectomy tissue and LuCaP 86.2 castration-resistant PDX-derived organoids.
CPT1A variant expression with area measurements through day 15; a separate ASO comparison in LuCaP 86.2 organoids.
Models
Organoids from fresh patient tissue and LuCaP 86.2 PDXO cultures; prostate cancer cell lines and benign RWPE-1 for distinct supporting assays.
Perturbations and measurements
CPT1A variant 1/variant 2 constructs; THRAP3-site ASO and scrambled control; organoid area, qRT-PCR, RNA immunoprecipitation and cell-line viability.
Variant-expression experiments use vector controls, and ASO experiments use a scrambled control. Patient-derived and PDX-derived organoids provide separate model comparisons.
Compare CPT1A variants in PDO/PDXO cultures and assess ASO response in LuCaP 86.2 PDXOs.
One LuCaP 86.2 PDX model and fresh treatment-naive patient tissue; the PDO growth assay measured 100 individual organoids per transduction group.
Empty-vector or CPT1A variant 1 versus variant 2; separate specific versus scrambled ASO comparisons.
CPT1A variant 2 promoted organoid growth, whereas variant 1 reduced growth. The ASO reduced LuCaP 86.2 organoid growth; separate cell-line RNA assays confirmed reduced CPT1A variant 2 and THRAP3 recruitment.
Citron kinase regulates THRAP3-dependent RNA splicing in prostate cancer models. An antisense oligonucleotide blocking THRAP3 recruitment to CPT1A pre-mRNA reduced growth in cell lines and a PDX-derived organoid model.
