Use of Positron Emission Tomography to Target Prostate Cancer Gene Therapy by Oncolytic Herpes Simplex Virus

Use of Positron Emission Tomography to Target Prostate Cancer Gene Therapy by Oncolytic Herpes Simplex Virus

polyDNA’s latest survey found that most male respondents did not know that genital herpes (HSV-2) can cause prostate cancer. The area of the skin affected by shingles is often very painful as infected nerves are inflamed. It is approximately 3 cm long and is the largest accessory gland of the male reproductive system. The seroprevalence of HHV-8 is higher in groups at risk for sexually transmitted diseases than in those not at risk (5). Canker sores occur only inside the mouth, on the tongue, and on the soft palate (roof of mouth), not on skin surfaces. In the last decade there has been a six-fold increase in diagnosis amongst forty-something men. This proof-of-concept study provides the mechanistic basis for selecting patients for targeted oncolytic viral therapy by means of a noninvasive molecular imaging method in the treatment of prostate cancer.

“Two types of antiviral treatments against HSV are available: topical and oral. The researchers did not take a dietary history but they considered it unlikely the high levels came just from eating a lot of oily fish (which are high in omega-3s) and are more likely to have come from supplements. It is mainly fibromuscular and contains very little glandular tissue. The study group included 10 men recruited through advertisements and physician referrals, and the study protocol was approved by the University of Washington Human Subjects Committee. Oral herpes (HSV-1) infection or exposure without noticeable infection is common. At CANCERactive we have been arguing this for 12 years. A screening method allowing us to predict which prostate cancer patients are likely to respond to oncolytic viral therapy would be beneficial in targeting HSV therapy to selected patients.

In a post marketing clinical study published on August 12, 2013 in the medical journal Pharmacology & Pharmacy in a special edition on Advances in Antiviral Drugs, researchers showed that Gene-Eden-VIR is safe and effective (2). PET is also under investigation for the evaluation of response to treatment (17), and to monitor the delivery of transduced genes during gene therapy (18;19). One of the problems is Hypertrophy of the Prostate, which is an enlargement of the prostate and is common after middle age. Semen, saliva, and blood were obtained prior to prostate biopsy. Mouth sores most commonly occur in children 1-2 years of age, but they can affect people at any age and any time of the year also termed herpes gingivostomatitis •    People contract HSV-1 by touching infected saliva, mucous membranes, or skin. He has conducted clinical trial research into this himself and has launched a product called POMI-T containing a number of Bioactive natural compounds like polyphenols. We therefore hypothesized that FDG-PET can be used as a surrogate marker for predicting tumor response to oncolytic HSV therapy in a prostate cancer xenograft model.

The androgen-sensitive prostate cancer cell line CWR22 (a kind gift of Dr. Zohar Dotan, Memorial Sloan Kettering Cancer Center (MSKCC), New York, NY) was grown in phenol red-free RPMI supplemented with 2mM L-glutamine, 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, 1.0 mM sodium pyruvate, 100 U/ml penicillin, 100 mg/ml streptomycin, and 10% charcoal dextran treated fetal bovine serum (Atlanta Biologicals, Norcross, GA). According to the American Cancer Society, this year approximately 39,000 men will die of this disease and 184,500 new cases of prostate cancer will be diagnosed. KS1 and KS2 primers were used to amplify the KS330 Bam233 fragment of ORF 26 (9). Asymptomatic infection occurs twice as often as the disease with symptoms. And considering all the side-effects of orthodox cancer treatment, this is research that should be heeded. Viral stocks were propagated on Vero cells (American Type Culture Collection, Rockville, MD) and titered by standard plaque assay.

CWR22 cells were plated (5 x 104 cells per well and 2 x 104 cells per well, respectively) in 12-well flat-bottom plates with either 3 μM DHT (dihydrotestosterone) or vehicle alone. After incubation for 12 hours, each well was infected with NV1066 at a multiplicity of infection (MOI, or the ratio of viral plaque forming units [PFU] per tumor cell) of 0.005 and 0.01. A physician inserts a gloved finger into the anus and digitally palpates the posterior lobe of the prostate. The resulting DIG-labeled transcripts were precipitated with ethanol and quantified spectrophotometrically. •    Herpes simplex virus (type 1, herpes-1or HSV-1), which causes about 80% of cases of oral herpes infections •    Other herpes simplex viruses (type 2, herpes-2 or HSV-2), which causes the other approximately 20% These viruses enter the body through small cuts, abrasions, or breaks in the skin or mucous membranes. Nothing could be further from the truth. Viable cells from individual wells were counted after trypsinization and staining with trypan blue.
Use of Positron Emission Tomography to Target Prostate Cancer Gene Therapy by Oncolytic Herpes Simplex Virus

All samples were analyzed in triplicate. All animal procedures were performed with the approval of the Memorial Sloan-Kettering Institutional Animal Care and Use Committee. For example, the most common treatment offered to patients with localized prostate cancer and a 10-year or greater life expectancy is radical prostatectomy. Hybridized probe was detected with alkaline-phosphatase-conjugated anti-DIG antibodies (1,500 mU/ml) and nitroblue tetrazolium–5-bromo-4-chloro-3-indolylphosphate toluidinium substrate, following the manufacturer’s recommendations (Boehringer Mannheim). The HSV viruses multiply in the human cells by overtaking and utilizing most of the human cells functions. A further complication is that after a high PSA reading, you´ll be rushed into a biopsy. In all experiments, tumors were measured three times weekly after implantation, and tumor volumes were recorded.

The fluorine-18 labeled 2-[18F]-fluoro-2-deoxy-D-glucose (18FDG) was prepared using the General Electric PETtrace Microlab. All radiopharmaceuticals were subjected to stringent quality control assays including sterility and apyrogenicity testing of batches 18FDG was formulated in phosphate buffered saline (PBS). This is designed to deliver irradiation from radioactive seeds placed directly into the prostate. Viral RNA was not detected in uninduced cells, which were shown by end-point PCR to have 70% of induced cells and >90% of cells in which HHV-8 DNA was amplified by PCR (see below) demonstrated HHV-8 by in situ hybridization with both the T0.7 and T1.1 riboprobes, indicating that >50 HHV-8 copies/cell were required for detection by the RNA probes. It is used only for people with weakened immune systems, infants younger than 6 weeks of age, or people with severe disease. The great majority are slow growing and ´may be left for ten years or more´. Rats were positioned prone and a 2 minute scout transmission was performed to verify correct positioning.

2.2 mCi 18FDG in 100 μL PBS was then injected into the penile vein over 30 seconds and whole-body activity was recorded. The PET dynamic scan consisted of 10 one minute frames, 10 two minute frames, and 3 five minute frames. The patient’s quality of life and life expectancy, ignoring the fact that they have prostate cancer, need to be addressed. After 25 cycles of denaturation at 94°C for 1 min and annealing at 55°C for 2 min, followed by polymerization for 2 min at 72°C, the slides were treated for 5 min with xylenes to remove mineral oil, 5 min in 100% ethanol, and then air dried. Most investigators suggest consulting an infectious disease expert when HSV infected people need hospitalization. Several genes have been identified (including BRCA 1 and BRCA 2) which are linked to increased risk, and sometimes prostate cancer is thought to run in families. These same ROIs were then copied onto all dynamic frames, allowing the generation of physical decay corrected time activity curves.

Vendor provided software on the GE Advance scanner was used to calculate maximum standardized uptake values (SUVmax). All values were normalized to the SUVmax of both the liver and chest wall. This resulted in the death of the cancerous cells. All five men with HHV-8 DNA in prostatic tissue currently had or once had KS. Seven days after tumor implantation, animals were randomized to undergo either orchiectomy or sham-operation. Its function is connected with the whole urine and reproductive system. Two additional tumors from each group were treated with PBS alone to serve as a negative control.

Seventy-two hours after treatment, animals were sacrificed and tumors were harvested, weighed, and snap frozen in liquid nitrogen for subsequent analysis. Genomic DNA was isolated per standard protocol (DNeasy Tissue Kit, Qiagen Inc., Valencia, CA). These results suggest that the gene is involved in prostate cancer. 11 to 14) exhibited HHV-8 by either technique. Forward (5’-GTAACCCGTTGAACCCCATT-3’) and reverse (5’-CCATCCAATCGGTAGTAGCG-3’) primers were also designed for an 87-bp coding sequence of the 18S housekeeping gene to normalize the amount of DNA present. Even then there may be no further distress for ten or more years. Total DNA was isolated and used to construct a standard curve via serial dilutions.

A value of PFU per tumor was then obtained via comparison with standard curve. Quantitative PCR was repeated twice for tumor samples with similar results.

You may also like