Assessment of HER2 amplification status in breast cancer using a new automated HER2 IQFISH pharmDx™ (Dako Omnis) assay.

In breast cancer the human epidermal growth factor receptor 2 (HER2) is an important target for a number of different HER2 inhibitors. Different slide-based assays are available for assessment of treatment eligibility, which include fluorescence in situ hybridization (FISH) or other in situ hybridization (ISH) methods for assessment of the HER2 gene status. Here we report a summary of the validation data on HER2 IQFISH pharmDx™ (Dako Omnis), a newly developed assay for the automated staining platform Dako Omnis. The assay uses a non-toxic buffer that significantly reduces the hybridization time, which results in a total turnaround time of 3½ to 4h from deparaffinization to counting of the gene and centromere signals. The data reported in the current summary covers method comparison, assessment of staining quality, observer-to-observer reproducibility as well as reproducibility within and between laboratories. Based on data from the different studies it was concluded that HER2 IQFISH pharmDx (Dako Omnis) is a reliable and robust assay with a high precision that is at least comparable to the manual HER2 IQFISH pharmDx™ assay and the PathVysion(®)HER-2 DNA Probe Kit.

Analysis of HER2 status in gastroesophageal tumor specimens using a new automated HER2 IQFISH pharmDx™ (Dako Omnis) assay.

The human epidermal growth factor receptor 2 (HER2) is an important target for treatment of gastroesophageal cancer. Different slide-based assays are available for assessment of HER2 status. Overexpression of the HER2 protein is assessed by immunohistochemistry (IHC) whereas amplification of the HER2 gene is assessed by fluorescence in situ hybridization (FISH) or other in situ hybridization (ISH) methods. Here we report a summary of the validation data on HER2 IQFISH pharmDx™ (Dako Omnis), a newly developed assay for the automated staining platform Dako Omnis. This assay uses a non-toxic buffer that significantly reduces the hybridization time, which results in a total turnaround time of less than 4 hours from deparaffinization to counting of the gene and centromere signals. The data reported in the current summary cover method comparison, assessment of staining quality, observer-to-observer reproducibility as well as reproducibility within and between laboratories. Based on data from the different studies it was concluded that HER2 IQFISH pharmDx (Dako Omnis) is a reliable and robust assay, with high precision and at least comparable to the manual HER2 IQFISH pharmDx™ assay. The HER2 IQFISH pharmDx (Dako Omnis) assay is currently not commercially available outside the Europe Union.

Tea polyphenols and theaflavins are present in prostate tissue of humans and mice after green and black tea consumption.

Green and black tea have shown promise in the chemoprevention of prostate cancer. The objective of this study was to determine the bioavailability and bioactivity of tea polyphenols (PP) and theaflavins in human serum and human and mouse tissues. A decaffeinated black tea diet was administered to C57BL/6 mice. PPs and theaflavins were found in the small and large intestine, liver, and prostate in conjugated and free forms. The relative prostate bioavailability of theaflavin was 70% higher than that of epigallocatechin gallate (EGCG). In the second mouse study, a green tea (GT) diet was administered followed by the control diet for 1-5 d. Epicatechin (EC), EGCG, and epicatechin gallate (ECG) concentrations in prostate tissue were significantly decreased after 1 d of consuming the control diet. Epigallocatechin gallate (EGC), however, did not decrease significantly. For the human study, 20 men scheduled for surgical prostatectomy were randomly assigned to consume 1.42 L daily of GT, BT, or a caffeine-matched soda control (SC) for 5 d before radical prostatectomy. Tea PPs were greater in prostate samples from men consuming BT and GT than in men consuming SC (P = 0.0025). Although tea PP were not detectable in serum, ex vivo LNCaP prostate cancer cell proliferation was less when cells were grown in media containing patient serum collected after BT (P < 0.001) and GT (P = 0.025) consumption relative to baseline serum This is the first human study to show that tea polyphenols and theaflavins are bioavailable in the prostate where they may be active in the prevention of prostate cancer.

Clinical utility of percent prostate needle biopsy tissue with cancer cutpoints to risk stratify patients before radical prostatectomy.

OBJECTIVES: The percentage of total prostate needle biopsy tissue with cancer was previously found to be a stronger predictor of biochemical failure after radical prostatectomy (RP) than either biopsy Gleason score or serum prostate-specific antigen (PSA). To improve our ability to predict preoperatively the risk of biochemical recurrence after RP, we sought to determine the cutpoints of the percentage of biopsy tissue with cancer to separate patients into low, intermediate, or high-risk groups. We then examined whether we could further stratify low, intermediate, and high-risk groups (on the basis of the PSA level and biopsy Gleason score) using the percentage of prostate needle biopsy tissue with cancer. METHODS: A single pathologist reviewed the prostate needle biopsy specimens of 217 men who underwent RP between 1991 and 2001. Biopsy specimens were examined for Gleason score and the percentage of total biopsy tissue with cancer. Cutpoints were identified to define patients with differing risk of biochemical recurrence after RP. These cutpoints were applied to low, intermediate, and high-risk patients, on the basis of PSA and biopsy Gleason score, to determine whether preoperative risk stratification could be improved. RESULTS: Using the cutpoints for the percentage of prostate needle biopsy tissue with cancer of less than 20% (low risk), 20% to less than 55% (intermediate risk), and 55% or greater (high risk), patients were separated into three groups with differing risks of biochemical failure after RP (hazard ratio 1.95, 95% confidence interval 1.37 to 2.77, P <0.001). These cutpoints further stratified patients with an intermediate (P = 0.002) or high risk (P = 0.05) of biochemical failure (on the basis of the PSA and biopsy Gleason score). However, these cutpoints provided no improvement in risk stratification for patients who were at low risk (P = 0.501) of biochemical failure (on the basis of PSA and biopsy Gleason score). CONCLUSIONS: The percentage of total prostate needle biopsy tissue with cancer can be used to stratify patients into low, intermediate, and high-risk groups preoperatively for biochemical recurrence after RP. These cutpoints could further stratify patients preoperatively who were at intermediate or high risk of biochemical failure on the basis of PSA and biopsy Gleason score.

Percent prostate needle biopsy tissue with cancer is more predictive of biochemical failure or adverse pathology after radical prostatectomy than prostate specific antigen or Gleason score.

PURPOSE: Biopsy Gleason score, serum prostate specific antigen (PSA) levels, and clinical stage are known to be independent predictors of adverse pathological features and biochemical failure after radical prostatectomy. We determine whether various prostate needle biopsy parameters were predictive of either adverse pathological findings or disease recurrence after radical prostatectomy. MATERIALS AND METHODS: A single pathologist reviewed the prostate needle biopsy specimens of 190 men who underwent radical prostatectomy between 1991 and 2000. Biopsy specimens were examined for Gleason score, perineural invasion, number and percent of cores with cancer, and percent of total biopsy tissue with cancer and Gleason grade 4 or 5 cancer. Multivariate analysis was used to determine the prostate needle biopsy parameters and preoperative clinical variables, including serum PSA, clinical stage, patient age and race, that were most significant for predicting positive surgical margins, nonorgan confined disease, seminal vesicle invasion and biochemical failure after radical prostatectomy. RESULTS: Of the prostate needle biopsy parameters examined percent of tissue with cancer was the strongest predictor of biochemical recurrence in the multivariate analysis (p <0.001). Percent of tissue with cancer was a stronger predictor of biochemical recurrence than either PSA (p = 0.048) or biopsy Gleason score (p = 0.053). It was also a strong independent predictor of seminal vesicle invasion (p = 0.015) and nonorgan confined disease (p = 0.024). Perineural invasion, percent and number of cores with cancer, and percent of tissue with Gleason grade 4 or 5 were not independent predictors of either adverse pathology or biochemical failure. CONCLUSIONS: Of all the preoperative variables examined, including the standard clinical variables of serum PSA, Gleason score and clinical stage, percent of biopsy tissue with cancer was the strongest predictor of biochemical recurrence, seminal vesicle invasion and nonorgan confined disease. Consideration should be given to reporting percent of total biopsy tissue with cancer in all prostate biopsy results.

Preoperative p27 status is an independent predictor of prostate specific antigen failure following radical prostatectomy.

PURPOSE: p27 is an important cell cycle regulator, and decreased expression in radical prostatectomy specimens is associated with an increased risk of prostate specific antigen (PSA) failure. To our knowledge no prior study has shown that preoperative p27 status independently predicts recurrence after radical prostatectomy. MATERIALS AND METHODS: The prostate needle biopsy specimens of 161 men treated with radical prostatectomy were examined for p27 expression using immunohistochemistry. Various p27 cut points were examined for their ability to separate patients into groups with different risk for time to biochemical recurrence following radical prostatectomy. The best p27 cut point was compared to other clinical variables (PSA, clinical stage, age, biopsy Gleason score and percent of prostate needle biopsy with cancer) on multivariate analysis to determine which variables independently predicted biochemical failure. RESULTS: A p27 cut point of less than 45% positive staining cells resulted in significant preoperative risk stratification for time to PSA failure (HR 2.41, p = 0.010). On multivariate analysis serum PSA (HR 1.04, p = 0.011), biopsy Gleason score (HR 1.51, p = 0.011), percent of biopsy tissue with cancer (HR 10.01, p = 0.001) and less than 45% p27 positive cells (HR 2.44, p = 0.014) were all independent predictors of biochemical recurrence. CONCLUSIONS: Preoperative p27 expression is an independent predictor of time to biochemical recurrence following radical prostatectomy. Patients with less than 45% p27 positive cells in the prostate needle biopsy specimen have almost a 2.5-fold increased risk of biochemical recurrence. To our knowledge this study is the first to show that p27 status of the prostate needle biopsy specimen can be used before radical prostatectomy to predict biochemical failure.

Comparison of percentage of total prostate needle biopsy tissue with cancer to percentage of cores with cancer for predicting PSA recurrence after radical prostatectomy: results from the SEARCH database.

OBJECTIVES: Tumor volume in the prostate needle biopsy is an important prognosticator for patients with prostate cancer. However, the best method to measure tumor volume in the prostate needle biopsy is unknown. We compared the total percentage of biopsy tissue with cancer to the percentage of cores positive for their ability to predict adverse pathologic findings and biochemical failure after radical prostatectomy (RP). METHODS: A retrospective survey of 355 patients from the Shared Equal Access Regional Cancer Hospital database treated with RP between 1990 and 2002 was undertaken. Multivariate analysis was used to compare the percentage of cores and percentage of tissue with cancer to the standard clinical variables of age, prostate-specific antigen (PSA) level, biopsy Gleason score, and clinical stage for their ability to predict positive surgical margins, non-organ-confined disease, seminal vesicle invasion, and time to PSA recurrence after RP. RESULTS: On multivariate analysis, the percentage of tissue with cancer significantly predicted non-organ-confined disease and seminal vesicle invasion, but the percentage of cores did not significantly predict any of the pathologic features examined. In separate multivariate analysis, only the percentage of tissue with cancer, but not the percentage of cores with cancer, significantly predicted PSA failure. Moreover, when compared in the same multivariate analysis, only the percentage of tissue with cancer (hazard ratio 8.25, 95% confidence interval 3.06 to 22.22, P <0.001) was a significant predictor. The area under the receiver operating curves for predicting PSA failure was significantly greater for the percentage of tissue with cancer (0.697) than for the percentage of cores (0.644, P = 0.022). Cutpoints for the percentage of tissue with cancer (less than 20%, 20% to 40%, and greater than 40%) and the percentage of cores (less than 34%, 34% to 50%, greater than 50%) both provided significant preoperative risk stratification for biochemical failure, although the percentage of tissue with cancer cutpoints provided better risk stratification (higher hazard ratios and lower P value). Cutpoints for the percentage of tissue with cancer but not the percentage of cores positive further stratified patients who were at low (P = 0.041), intermediate (P = 0.002), and high (P = 0.023) risk on the basis of the PSA level and biopsy Gleason score. CONCLUSIONS: The percentage of tissue with cancer was better than the percentage of cores at predicting advanced pathologic features and PSA recurrence after RP. Unlike the percentage of cores, the percentage of tissue with cancer cutpoints further stratified low, intermediate, and high-risk patients on the basis of PSA level and biopsy Gleason score. Although the percentage of tissue with cancer is a slightly more cumbersome measurement than the percentage of positive cores, it provided statistically and clinically superior preoperative risk stratification for biochemical failure after RP.

Preoperative model for predicting prostate specific antigen recurrence after radical prostatectomy using percent of biopsy tissue with cancer, biopsy Gleason grade and serum prostate specific antigen.

PURPOSE: We developed a preoperative model to risk stratify patients for prostate specific antigen (PSA) failure following radical prostatectomy (RP) and identify those at high risk who would be potential candidates for neoadjuvant clinical trials. MATERIALS AND METHODS: A retrospective survey of 459 patients from the SEARCH Database treated with RP between 1990 and 2002 was done. Multivariate analysis was used to compare the preoperative variables of patient age, race, PSA, biopsy Gleason score, clinical stage and percent of prostate needle biopsy tissue with cancer for the ability to predict time to PSA recurrence following RP. Significant independent predictors were combined to create a novel risk grouping model. RESULTS: On multivariate analysis biopsy Gleason score (p < 0.001), percent of biopsy tissue with cancer (p < 0.001) and serum PSA (p = 0.001) were the only significant independent predictors of PSA failure. Combining these 3 significant predictors of PSA failure using previously published cutoff points for each variable generated a 4 tier preoperative model for predicting biochemical failure following RP (HR 1.91 for each 1 risk category increase, CI 1.62 to 2.26, p < 0.001). The model further stratified patients who were already stratified into low, intermediate and high risk groups based on a previously described model using PSA, biopsy Gleason score and clinical stage. A simplified table was developed to predict the risk of biochemical recurrence within 2 years following surgery, as stratified by percent of tissue with cancer, PSA and biopsy Gleason score. CONCLUSIONS: A combination of serum PSA, biopsy Gleason score and percent of prostate biopsy tissue with cancer define a new preoperative model for predicting PSA failure following RP. This model further stratified patients who were already stratified based on PSA, biopsy Gleason score and clinical stage, and it can be used preoperatively to identify patients at high risk who would be candidates for neoadjuvant clinical trials. Using this model an easy to use table was developed to predict preoperatively the 2-year risk of PSA recurrence following RP.

Predicting biochemical recurrence after radical prostatectomy for patients with organ-confined disease using p27 expression.

OBJECTIVES: It is unclear why men who undergo radical prostatectomy (RP) and are found to have pathologically organ-confined disease develop prostate-specific antigen (PSA) recurrences. We previously found that patients with less than 45% of cells in the prostate needle biopsy specimen (PNBx) staining positive for the cell cycle regulator p27 had a significantly increased risk of biochemical recurrence after RP. We sought to determine whether p27 staining in the PNBx specimen might serve as a molecular marker for PSA failure in the subset of patients who develop PSA recurrence despite organ-confined disease at RP. METHODS: The PNBx specimens of 161 men treated with RP between 1991 and 2000 were examined for p27 expression using immunohistochemistry. The p27 cutpoint of less than 45% expression was used to define the high and low-risk categories. Patients were separated into two groups for analysis: organ-confined (pT2 and negative surgical margins) and non-organ-confined (pT2 with positive surgical margins, pT3, pT4, or lymph node involvement). The mean and median follow-up for patients with organ-confined and non-organ-confined disease was 47 and 43 months and 42 and 38 months, respectively. Multivariate Cox proportional hazards analysis was used to examine the preoperative clinical variables that were the strongest predictors of biochemical recurrence after RP among each group. RESULTS: Among organ-confined patients, p27 expression was the only significant independent predictor of the time to biochemical recurrence after RP (hazard ratio 5.15, 95% confidence interval 1.41 to 18.83, P = 0.013). Among patients with non-organ-confined disease, the percentage of biopsy tissue with cancer, biopsy Gleason score, and PSA level were independent predictors of PSA recurrence. p27 expression was not a significant independent predictor of PSA recurrence among men with non-organ-confined disease. CONCLUSIONS: p27 expression in the PNBx was a significant independent predictor of PSA failure for patients with pathologically organ-confined disease, but not for those with non-organ-confined disease. Patients with organ-confined disease but low p27 expression had a greater than fivefold risk of developing PSA recurrence than were men with high p27 expression, suggesting that p27 may be a molecular marker associated with micrometastatic disease at the time of RP.