The results of real-time brachytherapy for the management of low- and intermediate-risk prostate cancer in patients with prostate volumes up to 100 mL.

OBJECTIVES: • To report the results of real-time brachytherapy in the management of low-risk and intermediate-risk prostate cancer in patients with prostate volumes up to 100 mL, over a 6-year period. • To prospectively determine whether prostate volume influences the ability to achieve a quality implant and therefore impact upon prostate-specific antigen (PSA) relapse-free survival, and urinary and rectal toxicity. SUBJECTS AND METHODS: • In all, 216 men with localized prostate cancer were treated with real-time prostate brachytherapy using (125) I implants between November 2003 and December 2009. • Patient selection was based upon functional parameters; International Prostate Symptom Score (IPSS) and flowmetry. • Patients had computed tomography imaging at 1 month to assess post-implant dosimetry. PSA, IPSS and Radiation Therapy Oncology Group rectal toxicity scores were recorded prospectively over the follow-up period. • Patients with prostate volumes ≤50 mL and those with volumes >50 mL were compared. RESULTS: • Overall PSA relapse-free survival was 98.8%; 97.0% for intermediate-risk patients and 100.0% for low-risk patients. By volume, 98.5% of men with standard prostates were free from PSA relapse compared with 100.0% of men with large prostates. • The mean post-implant D90 was 177.0 Gy; 175.5 Gy in standard prostates and 183.5 Gy in large prostates. • The overall acute urinary retention rate was 1.9%; 1.7% in standard prostates and 2.4% in large prostates. There were three urethral strictures, all in the standard prostate group. The mean IPSS increased to 11 and 14 at 3 months for the standard and large prostate groups, respectively, before settling to 2 above baseline for both groups at 12 months. • There were no rectovesical fistulae. Persistent rectal bleeding was reported by one (0.5%) patient in the standard prostate group. CONCLUSIONS: • Prostate brachytherapy is effective in the treatment of low-risk and intermediate-risk prostate cancer. • It is technically possible to deliver a quality implant in a large prostate using real-time brachytherapy. • The treatment itself is well tolerated. Prostate volumes up to 100 mL should not exclude patients from brachytherapy providing either flow rate ≥14 mL/s or symptom score (IPSS) ≤ 10.

Nuclear localization of factor inhibitor hypoxia-inducible factor in prostate cancer is associated with poor prognosis.

PURPOSE: We determined the role of factor inhibiting hypoxia-inducible factor-1 in prostate cancer specimens. MATERIALS AND METHODS: A tissue microarray of 152 prostate cancers was constructed and stained for factor inhibiting hypoxia-inducible factor-1, hypoxia-inducible factor-1α and 2α, and glucose transporter 1 as a prototypical downstream target of hypoxia-inducible factor-1α. Correlation analysis was done between these variables, and between factor inhibiting hypoxia-inducible factor-1, and clinical and pathological variables, including prostate specific antigen as a surrogate of recurrence. RESULTS: Factor inhibiting hypoxia-inducible factor-1 was expressed in the cytoplasm and/or the nucleus in 86.5% of tumors, including exclusive cytoplasmic expression in 51.3% and exclusive nuclear expression in 5.3%. Any nuclear and exclusive expression of factor inhibiting hypoxia-inducible factor was associated with poor prognosis on univariate analysis (p = 0.007 and 0.042, respectively). On multivariate analysis men with nuclear expression in tumors were twice as likely to experience recurrence (p = 0.034). CONCLUSIONS: Factor inhibiting hypoxia-inducible factor-1 is widely expressed in prostate tumors. Its differential subcellular expression suggests that regulation of its expression is an important factor in the activity of the hypoxia-inducible factor pathway. Its modulation may help treat hypoxia-inducible factor driven aggressive prostate cancer.

A tie-over dressing for graft application in distal penectomy and glans resurfacing: the TODGA technique.

OBJECTIVES: • To describe a novel method of split-skin graft (SSG) fixation for neo-glans formation after distal penectomy for penile cancer and glans resurfacing for carcinoma in situ or lichen sclerosus (LS); the TODGA technique. • Rather than 'quilting' the graft onto the neo-glans, which requires up to 5 days bed rest, the tie-over method fixes the graft adequately enough to allow immediate patient mobilization. PATIENTS AND METHODS: • In all, 41 consecutive operations, with a follow-up of ≥ 12 months, were performed on 40 patients (mean age 62 years, range 32-83) from December 2000 to October 2008, where a SSG was applied to the raw glans or penile stump. • The protocol varied for the first 12 operations on 11 patients. The tie-over dressing was left in place for 6 (one patient) or 7 days (11) and various materials were used; paraffin gauze (one), expanded foam (five) and proflavine-soaked gauze (six). The first two patients had their dressing removed under general anaesthetic but all subsequent patients had their dressing removed on the ward. • The subsequent 29 operations used the same protocol where a proflavine-soaked gauze dressing was left undisturbed for 10 days. RESULTS: • In the original 11 patients, two required re-grafting. After this initial development period, we amended the technique to use stronger sutures and left the dressing undisturbed for 10 days. • In addition, we standardized the use of proflavin-soaked gauze, as we found it easy to apply and remove. Since we adopted this protocol, we have performed 29 operations over a 3-year period. • The cosmetic results were excellent with only one patient requiring re-grafting. The mean and median postoperative length of stay was 2 days. • One patient with a urethral squamous cell carcinoma associated with urethral and glans LS required a urethral dilatation to allow a check cystoscopy, and a further asymptomatic patient had a meatal dilatation in the clinic but meatal stenosis was otherwise not seen, with no patients requiring regular meatal dilatation. CONCLUSION: • The TODGA technique of SSG application and fixation allows immediate mobilization and reduces hospital stay whilst providing excellent cosmetic results with a high percentage of graft uptake.

Restoration of the Lost Obscured Coronal Sulcus: A Novel Surgical Technique for Penile Lichen Sclerosus with Severe Preputial Adhesions.

BACKGROUND: Lichen sclerosus (LS) may cause the glans and prepuce to become fused, making a standard circumcision impossible. Most authorities recommend excision of the fused area with glans resurfacing, although partial circumcision is often performed. OBJECTIVE: To evaluate an alternative technique that preserves the fused area and allows a complete circumcision without grafting. DESIGN, SETTING, AND PARTICIPANTS: Over 3 yr (January 2016-March 2018), 28 men (age 28-93 yr; mean 62 yr) underwent the restoration of lost obscured coronal sulcus (ROLOCS) procedure with over 1 yr of follow-up. Complications were reviewed retrospectively with an additional survey. SURGICAL PROCEDURE: The shaft skin is incised at the corona. Dartos is divided, which allows antegrade dissection just outside the fused glans membrane. The foreskin is removed and shaft skin sutured to dartos below the corona. MEASUREMENTS: Postoperative pain, aesthetic satisfaction, sexual enjoyment, glans sensation, and urinary symptoms were measured. RESULTS AND LIMITATIONS: There were no major complications. In all cases, the coronal sulcus was restored and the glans skin became soft without skin grafting. All were satisfied with the aesthetics. Of the patients, <70% experienced mild to low-moderate pain; 55% and 25% had, respectively, improved or reduced glans sensation; and 40% reported improved enjoyment of sex. Histology showed LS in all cases with squamous cell carcinoma in four, including three out of five patients who had previously undergone partial circumcision. Although this is the largest series reported yet, the numbers were too small for a meaningful statistical analysis. CONCLUSIONS: The ROLOCS operation offers an aesthetically superior alternative to partial circumcision and is easier to perform with less morbidity than skin grafting. PATIENT SUMMARY: The restoration of lost obscured coronal sulcus (ROLOCS) procedure provides an alternative to partial circumcision or circumcision with skin grafting when the foreskin is welded to the head of the penis (glans) due to lichen sclerosus. It produces a good cosmetic result, but the glans can be sore until it heals.

Urethrocutaneous fistula: preserve the tract and turn it inside out: the PATIO repair.

OBJECTIVE: To describe a novel technique for repairing penile urethrocutaneous (UC) fistula, by dissecting out the fistula tract, but instead of excising it, to preserve it and turn it inside out (PATIO); this creates a flap valve inside the urethral lumen that prevents the ingress of urine allowing the fistula to heal. PATIENTS AND METHODS: Five UC fistulae in four patients (two adults and two children) were repaired using the PATIO technique over a 4-year period. The two adults had developed the UC fistula after complex salvage hypospadias repair for failed surgery as children. In addition, three UC fistulae were recurrent after failed attempts to close them using conventional techniques, including the interposition of locally sourced 'waterproofing' subcutaneous tissue and, in one child, a rotational skin flap. One child had a difficult coronal UC fistula with little more than a skin bridge distally. RESULTS: Four operations were performed as day-cases with no catheter; one child was catheterized and kept in hospital overnight to prevent retention after a caudal anaesthetic. The catheter was removed, before discharge, the following morning. All five repairs were successful with no complications or recurrence, with a mean (range) follow-up of 18 (2-50) months. CONCLUSION: The PATIO technique is easy to perform and can be done as a day-case with no need for urethral catheterization. It does not preclude the use of other techniques to minimize the risk of recurrence, but so far these have not been necessary. As with all new operations, reported after only a few cases, caution must be used when interpreting the results and more studies are needed. Nevertheless, the concept makes sense and the early results are encouraging, particularly as it has been used on UC fistulae most liable to recur.

The chances of subsequent cancer detection in patients with a PSA > 20 ng/ml and an initial negative biopsy.

Transrectal ultrasound (TRUS)-guided prostate biopsy is known to carry a significant false-negative rate, leading some patients to have multiple biopsies. We investigated cancer detection rates in patients with a PSA > 20 ng/ml and a negative initial biopsy. We reviewed our database of 2396 TRUS-guided biopsies done between 1997 and 2002 in order to give a follow-up of at least 6 years. PSA, PSA density (PSAD), PSA velocity (PSAV), prostate volume, and DRE findings were analysed in relation to cancer status. Of the patients, 388 (16%) had a PSA > 20 ng/ml, including 99 (26%) with benign biopsies. Of those, 67 were rebiopsied, including 19 (28%) with cancer on the first rebiopsy and four (6%) on further biopsies. PSAD, DRE, and volume significantly differed between rebiopsied patients with and without cancer (p < 0.05). Patients who present with a PSA > 20 ng/ml and have an initial negative biopsy have a high chance of malignancy being detected on a second biopsy. However, if a second biopsy is also negative, then the chances of subsequent biopsies showing signs of cancer are very low if the DRE is normal and particularly if the PSAD is >0.35 ng/ml/cm3.

The role of cell-free DNA size distribution in the management of prostate cancer.

Cell-free DNA has been shown to have diagnostic potential in a number of malignant diseases. Recently, the integrity or size distribution of these fragments has also been identified as having possible diagnostic value. The current study explores the role of this novel parameter in the clinical diagnosis of prostate cancer. Plasma samples, collected prospectively from men undergoing investigation for prostate cancer, were used to obtain a cell-free DNA sample. Real-time PCR was used to quantify the level of cell-free DNA (ng/ml) and its size distribution (delta CD in each case. Sixty-one samples were collected from patients with prostate cancer and 62 from those with benign histology. Analysis failed to reveal a statistically significant relationship between either the level of cell-free DNA (p = 0.82) or its size distribution (p = 0.91) and the presence of cancer. These results demonstrate that cell-free DNA is unlikely to be of diagnostic value in the clinical management of this disease.

Prospective study of quantitation of plasma DNA levels in the diagnosis of malignant versus benign prostate disease.

PURPOSE: The aim of this study was to determine the potential of cell-free DNA levels as a diagnostic marker for prostate cancer, having first established the effect that blood sample processing has on this measurement. EXPERIMENTAL DESIGN: A total of 152 blood samples were collected prospectively from patients before their prostate biopsy and 25 from men in two distinct control groups. Blood was processed to yield three components: one-spin plasma (n = 68), two-spin plasma (n = 152), and serum (n = 56) samples. RESULTS: Having established the effect of sample preparation on the measured DNA level, the more reliable two-spin plasma sample was used to determine the relationship between DNA and the presence of prostate cancer. Those patients with cancer (n = 78) had a significantly higher level of DNA compared with the control groups (P < 0.0001 and P < 0.0001). However, DNA levels in patients with a benign biopsy (n = 74) were significantly higher than the 78 patients confirmed to have cancer (P = 0.02). CONCLUSIONS: We conclude that the sample type used in the quantitation of cell-free DNA has an effect on the level reported. Elevated levels are present in the two-spin plasma samples of patients with prostate cancer compared with healthy controls but are not of diagnostic value during the management of prostate cancer.

The androgen receptor is significantly associated with vascular endothelial growth factor and hypoxia sensing via hypoxia-inducible factors HIF-1a, HIF-2a, and the prolyl hydroxylases in human prostate cancer.

PURPOSE: Hypoxia regulates key biological processes including angiogenesis via the transcription factor, hypoxia-inducible factor (HIF). In prostate cancer, angiogenesis is also influenced by androgens, and recent cell line studies suggest that this effect is partly mediated by HIF. The study aimed to assess whether a relationship exists in human prostate cancer between expression of the androgen receptor, HIFs, and the key angiogenesis factor, vascular endothelial growth factor (VEGF). EXPERIMENTAL DESIGN: A tissue microarray comprised of 149 radical prostatectomy specimens was constructed. Semiquantitative immunohistochemical analysis was used to assess the expression of the androgen receptor, VEGF and HIF-1a and 2a, and their regulatory prolyl hydroxylase enzymes (PHD1, PHD2, and PHD3). Statistical analysis compared these factors with each other and with prostate-specific antigen relapse. RESULTS: There was a significant correlation between HIF-1a and HIF-2a expression (P = 0.02), and with androgen receptor (P = 0.04 and P < 0.001, respectively) and VEGF expression (P = 0.05 and P < 0.001, respectively). VEGF was also significantly related to the androgen receptor (P = 0.05), whereas PHD2 was inversely related to HIF-2a expression. No significant association was shown between HIF-1a or HIF-2a and time to prostate-specific antigen recurrence (P = 0.20 and P = 0.94, respectively). CONCLUSIONS: These findings confirm the relationship between hypoxia and the androgen receptor in prostate cancer, and show for the first time, the role of HIF-2a in this disease process. They provide clinical evidence to support the recent cell line findings that androgens may regulate VEGF levels through the activation of HIF in androgen-sensitive tumors. Inhibition of both the HIF pathways may provide new therapeutic options in the management of this disease.

CSN1 Somatic Mutations in Penile Squamous Cell Carcinoma.

Other than an association with HPV infection, little is known about the genetic alterations determining the development of penile cancer. Although penile cancer is rare in the developed world, it presents a significant burden in developing countries. Here, we report the findings of whole-exome sequencing (WES) to determine the somatic mutational landscape of penile cancer. WES was performed on penile cancer and matched germline DNA from 27 patients undergoing surgical resection. Targeted resequencing of candidate genes was performed in an independent 70 patient cohort. Mutation data were also integrated with DNA methylation and copy-number information from the same patients. We identified an HPV-associated APOBEC mutation signature and an NpCpG signature in HPV-negative disease. We also identified recurrent mutations in the novel penile cancer tumor suppressor genes CSN1(GPS1) and FAT1 Expression of CSN1 mutants in cells resulted in colocalization with AGO2 in cytoplasmic P-bodies, ultimately leading to the loss of miRNA-mediated gene silencing, which may contribute to disease etiology. Our findings represent the first comprehensive analysis of somatic alterations in penile cancer, highlighting the complex landscape of alterations in this malignancy. Cancer Res; 76(16); 4720-7. ©2016 AACR.

Epigenetics markers of metastasis and HPV-induced tumorigenesis in penile cancer.

PURPOSE: Penile cancer is a rare malignancy in the developed world with just more than 1,600 new cases diagnosed in the United States per year; however, the incidence is much higher in developing countries. Although HPV is known to contribute to tumorigenesis, little is known about the genetic or epigenetic alterations defining penile cancer. EXPERIMENTAL DESIGN: Using high-density genome-wide methylation arrays, we have identified epigenetic alterations associated with penile cancer. Q-MSP was used to validate lymph node metastasis markers in 50 cases. A total of 446 head and neck squamous cell carcinoma (HNSCC) and cervical squamous cell carcinoma (CESCC) samples were used to validate HPV-associated epigenetic alterations. RESULTS: We defined 6,933 methylation variable positions (MVP) between normal and tumor tissue, which includes 997 hypermethylated differentially methylated regions associated with tumor supressor genes, including CDO1, AR1, and WT1. Analysis of penile cancer tumors identified a 4 gene epi-signature which accurately predicted lymph node metastasis in an independent cohort (AUC of 89%). Finally, we explored the epigenetic alterations associated with penile cancer HPV infection and defined a 30 loci lineage-independent HPV specific epi-signature which predicts HPV status and survival in independent HNSCC, CESC cohorts. Epi-signature-negative patients have a significantly worse overall survival [HNSCC P = 0.00073; 95% confidence interval (CI), 0.021-0.78; CESC P = 0.0094; HR = 3.91, 95% CI = 0.13-0.78], HPV epi-signature is a better predictor of survival than HPV status alone. CONCLUSIONS: These data demonstrate for the first time genome-wide epigenetic events involved in an aggressive penile cancer phenotype and define the epigenetic alterations common across multiple HPV-driven malignancies.

Expression of BNIP3 correlates with hypoxia-inducible factor (HIF)-1alpha, HIF-2alpha and the androgen receptor in prostate cancer and is regulated directly by hypoxia but not androgens in cell lines.

BACKGROUND: BNIP3 is a hypoxia-induced protein involved in cell death and survival but its role in human tumors is unclear. This study investigated the role of BNIP3 in prostate cancer. METHODS: The expression of BNIP3, the androgen receptor (AR), hypoxia inducible factor (HIF)-1alpha, HIF-2alpha and the hypoxia regulated gene GLUT1 were assessed in tissue microarrays constructed from 149 radical prostatectomy specimens. Statistics compared expression of these factors between each other, conventional clinicopathological parameters and PSA recurrence. Since an association between BNIP3 and AR and the HIFs was observed, the influence of hypoxia, dihydrotestosterone and the AR blocker, Casodex, was also investigated in prostate cell lines. RESULTS: BNIP3 was expressed in the nucleus and cytoplasm. Eight of 149 (5.5%) tumors showed no expression, 44/149 (29.5%) cases showed exclusively cytoplasmic expression, 17/149 (11.5%) cases showed exclusively nuclear expression and 80/149 (53.5%) cases showed both cytoplasmic and nuclear expression. There was a significant correlation between cytoplasmic BNIP3 expression and Gleason score (P=0.005), age (P=0.02), AR (P=0.001), and GLUT1 (P=0.006). There was a significant correlation between nuclear BNIP3 expression and HIF-1alpha expression (P=0.006) and HIF-2alpha expression (P=0.013) but no correlation between BNIP3 and pre-operative PSA, tumor volume, margin positivity or capsular invasion (all P>0.05). There was an increase in BNIP3 expression under conditions of hypoxia (0.1% 0(2)) but not with dihydrotestosterone stimulation or with Casodex treatment. CONCLUSIONS: These findings suggest that BNIP3 is directly regulated by hypoxia but that there may be a hormonal independent mechanism coordinating the expression of BNIP3 in prostate tumors.

Expression of the forkhead transcription factor FOXP1 is associated both with hypoxia inducible factors (HIFs) and the androgen receptor in prostate cancer but is not directly regulated by androgens or hypoxia.

BACKGROUND: FOXP1 is a member of the winged helix or forkhead transcription factors. Recent studies have indicated possible roles for FOXP1 as a candidate tumor suppressor gene and a potential estrogen receptor (ER) co-regulator in the development of breast cancer. This study investigated whether FOXP1 has a similar relationship to the androgen receptor (AR) in prostate cancer and how these factors relate to the presence of hypoxia. METHODS: FOXP1, the AR and various hypoxia-regulated proteins (HIF-1alpha, HIF-2alpha, and VEGF) were measured with immunohistochemistry using a tissue microarray constructed from 167 archival radical prostatectomies. Statistical analyses compared the co-expression of these factors both with each other and conventional parameters including patient age, pre-operative prostate specific antigen (PSA), post-operative Gleason score, capsular invasion, surgical margin status, tumor volume, and PSA recurrence. The influence of hypoxia, dihydrotestosterone, and the AR blocker Casodex was investigated in prostate cell lines VCaP and LNCaP in vitro. RESULTS: Expression of nuclear FOXP1 was significantly positively correlated with AR (P = 0.0001), hypoxia inducible factor 1alpha (HIF-1alpha) (P = 0.01), HIF-2alpha (P = 0.0001), and vascular endothelial growth factor (VEGF) (P = 0.007) expression. A positive significant relationship was also identified with the post-operative Gleason score (P = 0.03) but not with the other variables, including PSA recurrence (P > 0.05). There was no significant change in expression in FOXP1 protein levels under conditions of hypoxia (0.1%), dihydrotestosterone stimulation (10 or 100 nM), or androgen blockade with Casodex (1, 10, or 50 microM). CONCLUSION: These findings suggest that there may be a hormonal and hypoxia independent regulatory mechanism coordinating the expression of HIFs, the AR, and FOXP1 in prostate tumors.

The number of regulatory T cells in prostate cancer is associated with the androgen receptor and hypoxia-inducible factor (HIF)-2alpha but not HIF-1alpha.

BACKGROUND: Regulatory T cells (T(R)) mediate peripheral immunological tolerance and are implicated in tumor progression. Because prostate cancer is being investigated for treatment by immunotherapy, we have assessed tumor T(R) in prostate cancers. METHODS: T(R) cells were identified by FOXP3 in tissue microarrays (TMAs) from 146 radical prostatectomies and correlated with clinicopathological tumor parameters and prostatic specific antigen rise (PSA). RESULTS: Twenty of 146 tumors contained no T(R). The mean of the average for the remaining 146 patients was 7.24. There was a significant correlation between T(R) and androgen receptor (P=0.003) and with hypoxia-inducible factor (HIF)-2alpha (P=0.007) but not HIF-1alpha (P=0.25). There was no significant correlation between T(R) numbers and stage, capsular invasion, urethral margins, vascular invasion, Gleason score, pre-operative PSA, or time to PSA recurrence (all P>0.05). CONCLUSIONS: T(R) in prostate tumors shows significant heterogeneity and may be the result of hormonal and hypoxic signaling. Targeting these may reduce T(R) in tumors allowing more successful immune therapies.

A seven-year follow-up of men following a benign prostate biopsy.

OBJECTIVES: To determine the incidence and clinical relevance of newly diagnosed cases of prostate cancer in a group of men who had an elevated PSA and benign prostate biopsy 7 years previously. PATIENTS AND METHOD: Patients under the age of 80 years with an elevated PSA who had had a benign prostate biopsy in the 12 months between March 1, 1994 and February 28, 1995 were studied. One hundred and sixty four patients with a mean age of 66.8 years (range 47-79 years) were identified. The mean PSA for this group was 10.3ng/ml (range 4.1-81ng/ml). One hundred and fifty nine of the 164 (97%) hospital records were available for review and all but 21 (12.8%) of the General Practitioners were contacted. RESULTS: Eighteen (11%) of the original 164 patients were subsequently diagnosed with prostate cancer, 2 died from their disease. CONCLUSIONS: In a population where the follow-up of patients with a benign biopsy was arranged on clinical grounds alone, 11% of the study group were diagnosed with prostate cancer during a seven-year follow-up. Although some of these cancers appear to be slow growing, most of those diagnosed in the initial follow-up period were deemed to be clinically significant and a small proportion progressed rapidly to metastases. All patients who have an elevated PSA, but benign biopsy, should undergo a period of PSA monitoring until it is clear that their PSA is not rising. We propose an initial intensive monitoring period to avoid missing those with clinically aggressive disease.