Ad5/35E1aPSESE4: A novel approach to marking circulating prostate tumor cells with a replication competent adenovirus controlled by PSA/PSMA transcription regulatory elements

Abstract

Circulating tumor cells serve as useful biomarkers with which to identify disease status associated with survival, metastasis and drug sensitivity. Here, we established a novel application for detecting PSA/PSMA-positive prostate cancer cells circulating in peripheral blood employing an adenovirus called Ad5/35E1aPSESE4. Ad5/35E1aPSESE4 utilized PSES, a chimeric enhancer derived from PSA/PSMA promoters that is highly active with and without androgen. A fluorescence signal mediated by GFP expression upon Ad5/35E1aPSESE4 infection was selectively amplified in PSA/PSMA-positive prostate cancer cells in vitro and ex vivo. Furthermore, for the in vivo model, blood drawn from TRAMP was tested for CTCs with Ad5/35E1aPSESE4 infection and was positive for CTCs at week 16. Validation was performed on patient blood at various clinical stages and found out 1-100 CTCs expressing GFP upon Ad5/35E1aPSESE4 infection. Interestingly, CTC from one patient was confirmed to be sensitive to docetaxel chemotherapeutic reagent and to abundantly express metastasis-related genes like MMP9, Cofilin1, and FCER1G through RNA-seq. Our study established that the usage of Ad5/35E1aPSESE4 is effective in marking PSA/PSMA-positive prostate cancer cells in patient blood to improve the efficacy of utilizing CTCs as a biomarker.

Enhanced expression of FCER1G predicts positive prognosis in multiple myeloma

“…FCER1G inhibits the expression of certain Alzheimer's disease susceptibility genes by participating in Herpes simplex (HSV-1) escape strategy [9]. Interestingly, the abundant expression of FCER1G was found in the circulating tumor cells of a prostate cancer patient who was sensitive to docetaxel chemotherapeutic reagent [32]. In the PPI network, many important genes that were associated with MM had been screened.…”

Ninjurin1 Is Up-regulated in Circulating Prostate Tumor Cells and Plays a Critical Role in Prostate Cancer Cell Motility

“…Thus, the identification of novel markers specifically expressed on CTCs that are not down-regulated during EMT may increase the potential for utilizing CTCs in the clinical setting. To overcome the limitations associated with the high level of sensitivity required to isolate CTCs, we developed a novel technology using a replicationcompetent adenovirus, Ad5/5E1aPSESE4, to label PSA/PSMA(+) cells (7). Interestingly, this adenovirusbased diagnostic tool provided similar results as conventional image-guided or PSA-based predictions of disease progression in most patients, although several patients with local tumors harbored too few CTCs in the blood to facilitate this type of analysis.…”

“…For the separation of peripheral blood mononuclear cells (PBMCs) from whole blood, 5 ml of whole blood in a K2 EDTA tube was added to a 50 ml conical tube containing 4 ml of Ficoll-Paque PLUS (GE Healthcare Life Science, Pittsburg, PA, USA) and was gently mixed with PBS to reach a total volume of 10 ml. The PBMC sample was prepared in a 12-well plate as described (7). Then, PBMCs were infected with 0.01 multiplicity of infection (MOI) of Ad5/35E1aPSESE4 and determined whether to contain CTCs mediating green fluorescent protein (GFP) expression.…”

“…Analyzing genomic mutations in CTCs using established methods for cell enrichment and isolation, genomic amplification, library qualification and census-based sequencing has become a standard protocol for evaluating prostate cancer (6). We developed an alternative approach in which a replication-competent adenovirus is introduced exclusively into prostatespecific antigen/prostate-specific membrane antigen (PSA/PSMA)(+) cells, with this method not being dependent on EpiCAM or specific membrane proteins (7). Interestingly, this approach enabled us to detect circulating prostate cancer cells from several patients who had not been clinically diagnosed with metastatic tumor nodules.…”

Targeting strategies of adenovirus-mediated gene therapy and virotherapy for prostate cancer

“…Evaluation of the cytotoxic activity demonstrated that Ad5/35PSES.mRFP/ttk killed LNCaP and CWR22rv cells more effectively. In addition, the chimeric oncolytic adenovirus Ad5/35E1aPSESE4 also effectively killed PSA/PSMA-positive PCa cells in the peripheral circulation ( 112 ).…”