Raheela Ashfaq

Publications (113)487.28 Total impact

• Article: Risk stratification of organ confined bladder cancer after radical cystectomy using cell cycle related biomarkers.

  Shahrokh F Shariat, Thomas F Chromecki, Eugene K Cha, Pierre I Karakiewicz, Maxine Sun, Yves Fradet, Hendrik Isbarn, Douglas S Scherr, Patrick J Bastian, Karl Pummer, Harun Fajkovic, Arthur I Sagalowsky, Raheela Ashfaq, Matthias Doblinger, Richard J Cote, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: We tested whether assessing the expression of cell cycle related proteins (p53, pRB, p21 and p27) could predict clinical outcomes after radical cystectomy in patients with organ confined urothelial carcinoma of the bladder. Our study included a development cohort of 272 patients and an external testing cohort of 52 patients with chemotherapy naïve pT1-2N0M0 urothelial carcinoma of the bladder treated with radical cystectomy. Immunohistochemical staining of p53, p27, p21 and pRB was performed on the development cohort of 272 patients and the external testing cohort of 52 patients. Overall 260 (80.2%) patients had altered expression of at least 1 molecular marker and 105 (32.4%), 95 (29.3%), 44 (13.6%) and 16 (4.9%) had 1 to 4 altered molecular markers, respectively. Addition of the number of altered molecular markers increased the predictive accuracy of the base model for disease recurrence and cancer specific mortality by 15.6% and 14.8%, respectively (p <0.001). The base model included age, gender, pT1 vs pT2 stage, grade, number of lymph nodes removed, lymphovascular invasion and concomitant carcinoma in situ. The combination of molecular markers yielded a predictive accuracy superior to that of any single molecular marker. We developed nomograms for the prediction of recurrence-free and cancer specific survival. Assessment of the number of altered cell cycle regulatory proteins in the cystectomy specimen improves the prediction of urothelial carcinoma of the bladder recurrence and survival in patients with organ confined disease. A combination of multiple markers is needed to capture the complex biological behavior of urothelial carcinoma of the bladder.
  The Journal of urology 12/2011; 187(2):457-62. · 4.02 Impact Factor
• Article: Prognostic value of insulin-like growth factor II mRNA binding protein 3 in patients treated with radical prostatectomy.

  Thomas F Chromecki, Eugene K Cha, Karl Pummer, Douglas S Scherr, Ashutosh K Tewari, Maxine Sun, Harun Fajkovic, Claus G Roehrborn, Raheela Ashfaq, Pierre I Karakiewicz, Shahrokh F Shariat
  [show abstract] [hide abstract]
  ABSTRACT: Study Type - Prognosis (case series) Level of Evidence 4 What's known on the subject? and What does the study add? Insulin-like growth factor II mRNA binding protein 3 (IMP3) is associated with poor outcomes in a variety of malignancies. The role of IMP3 in protate cancer remains poorly understood. IMP3 expression was associated with features of aggressive biology and aggressive prostate cancer recurrence after surgery. Although IMP3 is differentially expressed in patients with features of biologically aggressive prostate cancer, it does not have independent prognostic value in patients treated with RP. To evaluate the association of insulin-like growth factor II mRNA binding protein 3 (IMP3) with pathological features and outcomes in patients treated with radical prostatectomy (RP). Immunohistochemical staining for IMP3 was performed on archival tissue microarray specimens from 232 consecutive patients treated with RP for clinically localized disease. None of the patients received neoadjuvant or adjuvant radiation or hormone therapy. IMP3 expression was histologically categorized as normal or abnormal. Disease recurrence was classified as aggressive if metastases were present, post-recurrence prostate-specific antigen (PSA) doubling time was less than 10 months, or if the patients failed to respond to salvage local radiation therapy. The median follow-up was 69.8 months (interquartile range [IQR]: 40.1-99.5). IMP3 expression was abnormal in 42 (18.1%) of 232 patients. IMP3 expression was associated with extracapsular extension (P= 0.020), seminal vesicle invasion (P= 0.024), lymphovascular invasion (P= 0.036) and a high pathological Gleason score (P= 0.009). The 5-year PSA recurrence-free survival for IMP3-negative patients was 83% (standard error [SE]= 3) vs 67% (SE = 8) in IMP3-positive patients (log-rank test, P= 0.015). In a multivariable analysis that adjusted for the effects of surgical margins, extracapsular extension and seminal vesicle invasion, PSA (hazard ratio [HR]: 1.04, P= 0.013), lymph node metastasis (HR: 16.7, P < 0.001) and a high pathological Gleason score (HR 4.3, P= 0.008) were significantly associated with PSA recurrence-free survival, whereas IMP3 expression was not (P= 0.11). Similarly, IMP3 expression was only associated with aggressive recurrence (HR 3.2, P= 0.006). IMP3 expression is abnormal in approximately one-fifth of prostate cancers. Although IMP3 is differentially expressed in patients with features of biologically aggressive prostate cancer, it does not have an independent prognostic value in patients treated with RP.
  BJU International 11/2011; 110(1):63-8. · 2.84 Impact Factor
• Article: Tamoxifen downregulates ets oncogene family members ETV4 and ETV5 in benign breast tissue: implications for durable risk reduction.

  David Euhus, Dawei Bu, Xian-Jin Xie, Venetia Sarode, Raheela Ashfaq, Kelly Hunt, Weiya Xia, Joyce O'Shaughnessy, Michael Grant, Banu Arun, William Dooley, Alexander Miller, David Flockhart, Cheryl Lewis
  [show abstract] [hide abstract]
  ABSTRACT: Five years of tamoxifen reduces breast cancer risk by nearly 50% but is associated with significant side effects and toxicities. A better understanding of the direct and indirect effects of tamoxifen in benign breast tissue could elucidate new mechanisms of breast carcinogenesis, suggest novel chemoprevention targets, and provide relevant early response biomarkers for phase II prevention trials. Seventy-three women at increased risk for breast cancer were randomized to tamoxifen (20 mg daily) or placebo for 3 months. Blood and breast tissue samples were collected at baseline and posttreatment. Sixty-nine women completed all study activities (37 tamoxifen and 32 placebo). The selected biomarkers focused on estradiol and IGFs in the blood; DNA methylation and cytology in random periareolar fine-needle aspirates; and tissue morphometry, proliferation, apoptosis, and gene expression (microarray and reverse transcriptase PCR) in the tissue core samples. Tamoxifen downregulated Ets oncogene transcription factor family members ETV4 and ETV5 and reduced breast epithelial cell proliferation independent of CYP2D6 genotypes or effects on estradiol, ESR1, or IGFs. Reduction in proliferation was correlated with downregulation of ETV4 and DNAJC12. Tamoxifen reduced the expression of ETV4- and ETV5-regulated genes implicated in epithelial-stromal interaction and tissue remodeling. Three months of tamoxifen did not affect breast tissue composition, cytologic atypia, preneoplasia, or apoptosis. A plausible mechanism for the chemopreventive effects of tamoxifen is restriction of lobular expansion into stroma through downregulation of ETV4 and ETV5. The human equivalent of murine multipotential progenitor cap cells of terminal end buds may be the primary target.
  Cancer Prevention Research 07/2011; 4(11):1852-62. · 4.91 Impact Factor
• Article: Association of tumor-associated trypsin inhibitor (TATI) expression with molecular markers, pathologic features and clinical outcomes of urothelial carcinoma of the urinary bladder.

  Oliver Patschan, Shahrokh F Shariat, Daher C Chade, Pierre I Karakiewicz, Raheela Ashfaq, Yair Lotan, Kristina Hotakainen, Ulf-Håkan Stenman, Anders Bjartell
  [show abstract] [hide abstract]
  ABSTRACT: PURPOSES: To describe the differential tissue expression of tumor-associated trypsin inhibitor (TATI) in normal bladder urothelium, primary urothelial carcinoma of the bladder (UCB) and metastatic UCB and to assess the association of TATI expression with molecular markers commonly altered in UCB and clinical outcomes after radical cystectomy. METHODS: Slides from eight cystectomy patients without cancer, 191 radical cystectomy patients, 20 lymph nodes without metastasis and 40 lymph nodes with UCB were stained. Tissue expression of TATI, cyclin E1, cyclin D1, p53, p21, p27, pRB, Ki-67, Bcl-2, Caspase-3, Survivin and Cyclooxigenase-2 was measured in a tissue microarray. Cancer-specific and recurrence-free survival after radical cystectomy was recorded. RESULTS: TATI was expressed in 100% of patients without cancer, while 71% of radical cystectomy specimens and 90% of lymph node metastases exhibited decreased or no TATI expression. In radical cystectomy specimens, TATI expression decreased with advancing pathologic stage (P < 0.001) and lymphovascular invasion (P = 0.055). In univariate analyses, but not in multivariable Cox proportional hazard regression analyses, decreased TATI expression was associated with increased probability of tumor recurrence and cancer-specific mortality. Decreased TATI expression was correlated with altered expression of Cyclooxigenase-2 (P = 0.005), p21 (P = 0.035) and Ki-67 (P = 0.004). CONCLUSIONS: We found that normal urothelium expresses TATI and that TATI expression decreases with advancing tumor stage. While there was no prognostic benefit to TATI when adjusted for standard clinicopathologic features, it seems to play an important biologic role in UCB pathogenesis and invasion. Its association with markers involved in the cell cycle, proliferation and inflammation serves as hypothesis for molecular interactions.
  World Journal of Urology 07/2011; · 2.41 Impact Factor
• Article: Microsatellite instability among individuals of Hispanic origin with colorectal cancer.

  Samir Gupta, Raheela Ashfaq, Payal Kapur, Bianca B Afonso, Thuy-Phuong T Nguyen, Faryal Ansari, C Richard Boland, Ajay Goel, Don C Rockey
  [show abstract] [hide abstract]
  ABSTRACT: Although the presence of microsatellite instability (MSI) in patients with colorectal cancer (CRC) may have implications for prognosis, therapy, and family counseling, to the authors' knowledge, the prevalence of MSI has not been well described among individuals of Hispanic origin with CRC residing in the United States. A retrospective cohort study using a hospital-based tumor registry to identify individuals of Hispanic origin who were diagnosed with CRC was conducted. Clinical data and tumor samples were retrieved. Molecular analyses included testing for MSI using a panel of 5 mononucleotide markers (BAT25, BAT26, NR21, NR24, and NR27) in a pentaplex polymerase chain reaction assay, as well as immunohistochemistry for the mismatch repair (MMR) proteins mutL homolog (MLH) 1, mutS homolog (MSH) 2, MSH6, and postmeiotic segregation increased 2 (PMS2) 2 on representative tissue. A total of 111 individuals of Hispanic origin with CRC were identified. Approximately 41.4% were women, and the median age was 57 years (interquartile range [IQR], 47.1-63.5 years). Eleven patients (9.9%; 95% confidence interval [95% CI], 4.2%-15.6%) had MSI CRC, whereas 14 patients (12.6%; 95% CI, 7.3%-21.8%) had CRC with ≥1 MMR protein abnormality. Ten of 11 individuals with MSI had clinical or molecular characteristics suspicious for Lynch syndrome such as abnormal expression of MSH2 and/or MSH6 (n=7) or age<50 years at the time of diagnosis (n=7). The prevalence of MSI CRC among Hispanic individuals may be similar to that of other races and ethnicities, but clinicopathological characteristics, including age at diagnosis and pattern of abnormal MMR protein expression, suggest that sporadic MSI CRC may be less common in individuals of Hispanic origin, and that much of the MSI observed in this situation may be attributable to Lynch syndrome. Further exploration of the causes of disparate presentations of CRC by ethnicity and race is warranted.
  Cancer 11/2010; 116(21):4965-72. · 4.77 Impact Factor
• Article: Loss of androgen receptor expression is not associated with pathological stage, grade, gender or outcome in bladder cancer: a large multi-institutional study.

  Carmen Mir, Shahrokh F Shariat, Theodorus H van der Kwast, Raheela Ashfaq, Yair Lotan, Andrew Evans, Sean Skeldon, Sally Hanna, Rati Vajpeyi, Cynthia Kuk, Sultan Alkhateeb, Juan Morote, Bas W G van Rhijn, Peter Bostrom, Jorge Yao, Hiroshi Miyamoto, Michael Jewett, Neil Fleshner, Ed Messing, Alexandre R Zlotta
  [show abstract] [hide abstract]
  ABSTRACT: • To investigate androgen receptor (AR) expression in a large series of patients with bladder cancer (BC) because data on a limited number of patients showed that loss of AR expression was associated with invasive BC. • A total of 472 patients with urothelial bladder carcinoma (UBC) from two institutional centres (Toronto and Dallas) were analysed. Tissue microarrays comprising both non-muscle-invasive UBC (n= 167) and muscle-invasive UBC (n= 305) were accrued and immunohistochemical staining for AR was performed. • We used bright-field microscopy imaging coupled with advanced colour detection software to detect, classify and count stained cellular objects and manual scoring. • Results obtained in Dallas were blindly reviewed and validated in Toronto and samples randomly chosen were further analysed in Rochester, NY, USA. • The AR were positively expressed in 61/472 (12.9%) bladder tumours. No statistically significant difference in AR expression between men and women was observed. • Only 9.0% of non-muscle-invasive BC expressed the AR compared with 15.1% of muscle-invasive tumours (P= 0.059). The highest percentage of AR positivity (28.9% of cases) was found in T2 tumours. • There was no statistically significant difference in death from BC, time to death, or time to recurrence between AR-positive and AR-negative cases. • In contrast to previous reports, based on our large BC series, we did not observe a decrease in AR protein expression in bladder tumours with increased pathological stage. Our data do not suggest that loss of AR expression is gender-related nor is it associated with invasive BC.
  BJU International 11/2010; 108(1):24-30. · 2.84 Impact Factor
• Article: Human epidermal growth factor receptor 2 expression status provides independent prognostic information in patients with urothelial carcinoma of the urinary bladder.

  Christian Bolenz, Shahrokh F Shariat, Pierre I Karakiewicz, Raheela Ashfaq, Richard Ho, Arthur I Sagalowsky, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: to test whether the expression of human epidermal growth factor receptor 2 (HER-2) is of prognostic value in a contemporary cohort of patients with urothelial carcinoma of the urinary bladder (UCB). tissue microarrays of 198 patients were constructed and immunohistochemical stainings were performed on the primary tumours and on lymphatic nodal metastases. All patients were treated with radical cystectomy (RC) and regional lymphadenectomy for UCB. HER-2 expression was assessed using continuous HER-2 expression scores (ranging from 0.1 to 3.9) generated using an automated cellular imaging system. Scores of ≥ 1.0 in at least 10% of tumour cells were regarded as HER-2 positive. We correlated HER-2 scores with pathological and clinical variables, including disease recurrence and cancer-specific mortality. of 198 patients undergoing RC with lymphadenectomy, there was HER-2 positivity in 55 primary tumours (27.8%) compared with 44.2% of the evaluable positive lymph nodes (P < 0.001). HER-2 positivity was significantly associated with the presence of lymphovascular invasion (LVI; P= 0.026). With a median (range) follow-up of 35.4 (1.3-176.1) months, 101 patients (51.0%) had UCB recurrence and 82 patients (41.4%) died from the disease. In multivariable analyses that adjusted for the effects of pathological tumour stage, grade, LVI, lymph node metastasis and adjuvant chemotherapy, HER-2 positive patients were at increased risk for both UCB recurrence (hazard ratio [HR] 1.955, P= 0.003) and UCB-specific mortality (HR 2.066, P= 0.004) compared with patients with negative HER-2 expression. a positive HER-2 status is associated with aggressive UCB and provides independent prognostic information for UCB recurrence and mortality. Assessment of HER-2 status can be used to identify patients at high risk of disease progression who may benefit from adjuvant HER-2-targeted mono- or combined therapy after RC.
  BJU International 10/2010; 106(8):1216-22. · 2.84 Impact Factor
• Article: Role of fluorescence in situ hybridization in bladder cancer surveillance of patients with negative cytology.

  Ramy F Youssef, Bruce J Schlomer, Richard Ho, Arthur I Sagalowsky, Raheela Ashfaq, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: The clinical utility of urine markers in urothelial cancer (UC) surveillance is not established. We previously evaluated the use of fluorescence in situ hybridization (FISH) in managing patients with atypical cytology at risk for UC. This study evaluates its role in patients with negative cytology with a history of UC. Between June 2007 and January 2009, every patient with a history of UC who underwent cystoscopy and cytology with UroVysion test were identified. A comprehensive chart review was performed on each patient with negative cytology. The population comprised 142 patients undergoing cancer surveillance; 111 patients with negative cystoscopy, 19 with equivocal cystoscopy, and 12 with positive cystoscopy. In patients with negative cystoscopy, there was cancer in only 1 of 111 patients. UroVysion could detect the only patient with UC with sensitivity of 100% and had a negative predictive value (NPV) of 100%. In patients with equivocal cystoscopy, it detected 2 tumors that would be missed by cytology. There were 4 false negative results (sensitivity 33.3% and NPV 66.7%). In patients with obvious lesion on cystoscopy, there were 9 false negative results (sensitivity 10% and NPV 18.2%). Few patients with negative cystoscopy and negative cytology have cancer. Patients with equivocal and positive cystoscopy and negative cytology frequently have cancer and the UroVysion FISH assay was not helpful in these cases. The cost-effectiveness of the FISH assay needs to be assessed prior to widespread use in patients with negative cytology.
  Urologic Oncology 05/2010; 30(3):273-7. · 3.22 Impact Factor
• Article: Association of angiogenesis related markers with bladder cancer outcomes and other molecular markers.

  Shahrokh F Shariat, Ramy F Youssef, Amit Gupta, Daher C Chade, Pierre I Karakiewicz, Hendrik Isbarn, Claudio Jeldres, Arthur I Sagalowsky, Raheela Ashfaq, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: We tested whether the altered immunohistochemical expression of angiogenesis related markers is associated with outcomes of patients with urothelial carcinoma of the bladder, and assessed the correlation of angiogenesis related markers with molecular markers commonly altered in urothelial bladder carcinoma. Vascular endothelial growth factor, basic fibroblast growth factor and thrombospondin 1 expression data were collected, as were microvessel density data. Immunohistochemical staining was performed on specimens from 204 patients treated with radical cystectomy for urothelial carcinoma of the bladder. We also stained serial sections of the specimens for cyclin E1, cyclin D1, p53, p21, p27, pRB, Ki-67, Bcl-2, caspase-3, survivin and cyclooxygenase-2. We measured time to disease recurrence and cancer specific mortality, as well as the association with clinical and pathological features and other molecular markers. The altered expression of vascular endothelial growth factor (over expression), basic fibroblast growth factor (over expression) and thrombospondin 1 (decreased expression) was 86%, 79% and 63%, respectively. Median microvessel density was 20. All 4 markers were associated with established clinicopathological features of aggressive urothelial carcinoma of the bladder (such as stage, lymphovascular invasion and lymph node metastasis) and other molecular markers. On multivariable analyses that adjusted for standard pathological features basic fibroblast growth factor and thrombospondin 1 were independent predictors of disease recurrence (HR 3.6, p = 0.002 and HR 2.2, p = 0.001, respectively) and cancer specific mortality (HR 2.8, p = 0.02 and HR 2.3, p = 0.003, respectively). When all 4 markers were included in 1 model basic fibroblast growth factor and thrombospondin 1 retained their independent association with disease recurrence (HR 2.9, p = 0.014 and HR 1.8, p = 0.022, respectively) and only thrombospondin 1 was independently associated with cancer specific mortality (HR 1.9, p = 0.031). Angiogenesis related molecular markers are commonly altered in urothelial carcinoma of the bladder, making them a target for therapy. Down-regulation of thrombospondin 1 and up-regulation of basic fibroblast growth factor are independent predictors of clinical outcomes of patients with urothelial carcinoma of the bladder.
  The Journal of urology 03/2010; 183(5):1744-50. · 4.02 Impact Factor
• Source

  Available from: Frank A Policht

  Article: Analysis of genetic copy number changes in cervical disease progression.

  Frank A Policht, Minghao Song, Svetlana Sitailo, Anna O'Hare, Raheela Ashfaq, Carolyn Y Muller, Larry E Morrison, Walter King, Irina A Sokolova
  [show abstract] [hide abstract]
  ABSTRACT: Cervical dysplasia and tumorigenesis have been linked with numerous chromosomal aberrations. The goal of this study was to evaluate 35 genomic regions associated with cervical disease and to select those which were found to have the highest frequency of aberration for use as probes in fluorescent in-situ hybridization. The frequency of gains and losses using fluorescence in-situ hybridization were assessed in these 35 regions on 30 paraffin-embedded cervical biopsy specimens. Based on this assessment, 6 candidate fluorescently labeled probes (8q24, Xp22, 20q13, 3p14, 3q26, CEP15) were selected for additional testing on a set of 106 cervical biopsy specimens diagnosed as Normal, CIN1, CIN2, CIN3, and SCC. The data were analyzed on the basis of signal mean, % change of signal mean between histological categories, and % positivity. The study revealed that the chromosomal regions with the highest frequency of copy number gains and highest combined sensitivity and specificity in high-grade cervical disease were 8q24 and 3q26. The cytological application of these two probes was then evaluated on 118 ThinPrep samples diagnosed as Normal, ASCUS, LSIL, HSIL and Cancer to determine utility as a tool for less invasive screening. Using gains of either 8q24 or 3q26 as a positivity criterion yielded specificity (Normal +LSIL+ASCUS) of 81.0% and sensitivity (HSIL+Cancer) of 92.3% based on a threshold of 4 positive cells. The application of a FISH assay comprised of chromosomal probes 8q24 and 3q26 to cervical cytology specimens confirms the positive correlation between increasing dysplasia and copy gains and shows promise as a marker in cervical disease progression.
  BMC Cancer 01/2010; 10:432. · 3.01 Impact Factor
• Article: Survivin as a prognostic marker for urothelial carcinoma of the bladder: a multicenter external validation study.

  Shahrokh F Shariat, Pierre I Karakiewicz, Guilherme Godoy, Jose A Karam, Raheela Ashfaq, Yves Fradet, Hendrik Isbarn, Francesco Montorsi, Claudio Jeldres, Patrick J Bastian, Matthew E Nielsen, Stefan C Müller, Arthur I Sagalowsky, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: The aim of the current study was to externally validate the value of survivin as a prognostic marker for bladder cancer in a large multi-institutional cohort of patients treated with radical cystectomy. Methods: The study comprised 726 patients treated with radical cystectomy and bilateral pelvic lymphadenectomy. Survivin staining and scoring were done with automated systems coupled with advanced color detection software. Specimens showing at least 10% reactivity were considered altered. Predictive accuracy was quantified using the concordance index and 200-bootstrap resamples were used to reduce overfit bias. Survivin was an independent predictor of disease recurrence and cancer-specific survival in multivariable analyses that controlled for the effects of standard clinicopathologic features (hazard ratios, approximately 1.6; P values < or = 0.002). In all patients (n = 726), addition of survivin to a model including standard clinicopathologic variables did not improve its predictive accuracy (P = 0.67 for disease recurrence and P = 0.27 for cancer-specific survival). In the subgroup of patients with pT(1-3)N(0)M(0) disease (n = 398), addition of survivin improved the accuracy of standard clinicopathologic features for prediction of disease recurrence and cancer-specific survival (1.3%, P < 0.001 and 1.2%, P < 0.001, respectively). Survivin expression improves our accuracy for prediction of cancer recurrence and survival in pT(1-3)N(0)M(0) patients by a small but statistically significant margin. Our findings support the need for further evaluation of survivin and its signaling pathways as well as survivin-targeted therapies in bladder cancer.
  Clinical Cancer Research 11/2009; 15(22):7012-9. · 7.74 Impact Factor
• Article: Combination of multiple molecular markers can improve prognostication in patients with locally advanced and lymph node positive bladder cancer.

  Shahrokh F Shariat, Daher C Chade, Pierre I Karakiewicz, Raheela Ashfaq, Hendrik Isbarn, Yves Fradet, Patrick J Bastian, Matthew E Nielsen, Umberto Capitanio, Claudio Jeldres, Francesco Montorsi, Seth P Lerner, Arthur I Sagalowsky, Richard J Cote, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: We tested whether the combination of 4 established cell cycle regulators (p53, pRB, p21 and p27) could improve the ability to predict clinical outcomes in a large multi-institutional collaboration of patients with pT3-4N0 or pTany Npositive urothelial carcinoma of the bladder. We also assessed whether the combination of molecular markers is superior to any individual biomarker. The study comprised 692 patients with pT3-4N0 or pTany Npositive urothelial carcinoma of the bladder treated with radical cystectomy and bilateral lymphadenectomy (median followup 5.3 years). Scoring was performed using advanced cell imaging and color detection software. The base model incorporated patient age, gender, stage, grade, lymphovascular invasion, number of lymph nodes removed, number of positive lymph nodes, concomitant carcinoma in situ and adjuvant chemotherapy. Individual molecular markers did not improve the predictive accuracy for disease recurrence and cancer specific mortality. Combination of all 4 molecular markers into number of altered molecular markers resulted in significantly higher predictive accuracy than any single biomarker (p <0.001). Moreover addition of number of altered molecular markers to the base model significantly improved the predictive accuracy for disease recurrence (3.9%, p <0.001) and cancer specific mortality (4.3%, p <0.001). Addition of number of altered molecular markers retained statistical significance for improving the prediction of clinical outcomes in the subgroup of patients with pT3N0 (280), pT4N0 (83) and pTany Npositive (329) disease (p <0.001). While the status of individual molecular markers does not add sufficient value to outcome prediction in patients with advanced urothelial carcinoma of the bladder, combinations of molecular markers may improve molecular staging, prognostication and possibly prediction of response to therapy.
  The Journal of urology 11/2009; 183(1):68-75. · 4.02 Impact Factor
• Article: Prospective validation of the clinical usefulness of reflex fluorescence in situ hybridization assay in patients with atypical cytology for the detection of urothelial carcinoma of the bladder.

  Bruce J Schlomer, Richard Ho, Arthur Sagalowsky, Raheela Ashfaq, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: We previously prospectively evaluated the clinical usefulness of a fluorescence in situ hybridization assay for treating patients with atypical cytology results at risk for bladder cancer. This study is a prospective validation the usefulness of fluorescence in situ hybridization in this setting. Between June 2007 and January 2009 every patient who underwent cystoscopy and cytology with atypical cytology underwent a reflex UroVysion test. A comprehensive review was then performed to evaluate clinical and pathological data on each patient. The study population comprised 108 patients with no history of cancer and 108 who underwent cystoscopy for cancer surveillance. In patients with cystoscopically visualized lesions UroVysion had a positive predictive value of 100% but there were false-negative results. In patients with equivocal cystoscopy and a history of cancer all 4 high grade tumors were detected and there were no false-negative findings. In patients with equivocal cystoscopy and no prior cancer the positive predictive value was 100% and there were no false-negative results. In patients with negative cystoscopy the UroVysion test detected all cancers but the positive predictive value was 10% and 29% in patients with and without a history of cancer, respectively. This prospective study of a reflex fluorescence in situ hybridization assay in patients with atypical cytology validates our previous findings. In patients with atypical cytology and obvious tumor on cystoscopy the assay was unnecessary but it was beneficial in those with equivocal or negative cystoscopy results. The fluorescence in situ hybridization assay identified all urothelial carcinoma tumors in patients with equivocal or negative cystoscopy. In patients with equivocal or negative cystoscopy and atypical cytology, a reflex fluorescence in situ hybridization assay may help avoid unnecessary evaluation while identifying those who would need further evaluation.
  The Journal of urology 11/2009; 183(1):62-7. · 4.02 Impact Factor
• Article: p53 predictive value for pT1-2 N0 disease at radical cystectomy.

  Shahrokh F Shariat, Yair Lotan, Pierre I Karakiewicz, Raheela Ashfaq, Hendrik Isbarn, Yves Fradet, Patrick J Bastian, Matthew E Nielsen, Umberto Capitanio, Claudio Jeldres, Francesco Montorsi, Stefan C Müller, Jose A Karam, Lukas C Heukamp, George Netto, Seth P Lerner, Arthur I Sagalowsky, Richard J Cote
  [show abstract] [hide abstract]
  ABSTRACT: Approximately 15% to 30% of patients with pT1-2N0M0 urothelial carcinoma of the bladder experience disease progression despite radical cystectomy with curative intent. We determined whether p53 expression would improve the prediction of disease progression after radical cystectomy for pT1-2N0M0 UCB. In a multi-institutional retrospective cohort we identified 324 patients with pT1-2N0M0 urothelial carcinoma of the bladder who underwent radical cystectomy. Analysis focused on a testing cohort of 272 patients and an external validation of 52. Competing risks regression models were used to test the association of variables with cancer specific mortality after accounting for nonbladder cancer caused mortality. In the testing cohort 91 patients (33.5%) had altered p53 expression (p53alt). On multivariate competing risks regression analysis altered p53 achieved independent status for predicting disease recurrence and cancer specific mortality (each p <0.001). Adding p53 increased the accuracy of multivariate competing risks regression models predicting recurrence and cancer specific mortality by 5.7% (62.0% vs 67.7%) and 5.4% (61.6% vs 67.0%), respectively. Alterations in p53 represent a highly promising marker of disease recurrence and cancer specific mortality after radical cystectomy for urothelial carcinoma of the bladder. Analysis confirmed previous findings and showed that considering p53 can result in substantial accuracy gains relative to the use of standard predictors. The value and the level of the current evidence clearly exceed previous proof of the independent predictor status of p53 for predicting recurrence and cancer specific mortality.
  The Journal of urology 08/2009; 182(3):907-13. · 4.02 Impact Factor
• Article: Estrogen and progesterone hormonal receptor expression in urothelial carcinoma of the bladder.

  Christian Bolenz, Yair Lotan, Raheela Ashfaq, Shahrokh F Shariat
  European urology 08/2009; 56(6):1093-5. · 7.67 Impact Factor
• Article: p53 expression in patients with advanced urothelial cancer of the urinary bladder.

  Shahrokh F Shariat, Christian Bolenz, Pierre I Karakiewicz, Yves Fradet, Raheela Ashfaq, Patrick J Bastian, Matthew E Nielsen, Umberto Capitanio, Claudio Jeldres, Jérôme Rigaud, Stefan C Müller, Seth P Lerner, Francesco Montorsi, Arthur I Sagalowsky, Richard J Cote, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: To test whether assessing p53 expression could improve the ability to predict disease recurrence and disease-specific survival in a multi-institutional cohort of patients with advanced urothelial carcinoma of the urinary bladder (UCB). The study comprised 692 patients with pT3-4 N0 or pTany N+ UCB treated with radical cystectomy and lymphadenectomy. The predictive accuracy (PA) was quantified using the 200 bootstrap-corrected concordance index. The base model comprised age, gender, stage, grade, lymphovascular invasion, number of lymph nodes removed, number of lymph nodes positive, concomitant carcinoma in situ, and adjuvant chemotherapy. p53 expression was altered in 341 (49.3%) patients. In multivariable analyses, p53 expression was independently associated with disease recurrence (hazard ratio, 1.66; P < 0.001) and cancer-specific mortality (hazard ratio 1.65, P < 0.001). Overall, adding p53 did not significantly improve the PA of the base model (recurrence +0.7%, P = 0.085, and cancer-specific mortality +1.2%, P = 0.050). In the subgroups of pT3N0 (280) and pT4N0 (83) patients, p53 slightly improved the PA of the base model by a statistically significant degree (recurrence +1.7% and +3.6%, respectively; cancer-specific mortality +1.9% and +3.5%, respectively; all P < 0.001). In 329 patients with pTany N+ disease p53 status did not improve the PA of the base model. While assessing p53 expression has limited utility in patients with lymph node-positive UCB, it marginally improves prognostication in patients with advanced non-metastatic UCB. Integration of p53 into a panel of biomarkers might be necessary to capture a more accurate picture of the biological potential of advanced UCB.
  BJU International 07/2009; 105(4):489-95. · 2.84 Impact Factor
• Source

  Available from: Claudio Jeldres Lanzarotti

  Article: Predictive value of combined immunohistochemical markers in patients with pT1 urothelial carcinoma at radical cystectomy.

  Shahrokh F Shariat, Christian Bolenz, Guilherme Godoy, Yves Fradet, Raheela Ashfaq, Pierre I Karakiewicz, Hendrik Isbarn, Claudio Jeldres, Jérôme Rigaud, Arthur I Sagalowsky, Yair Lotan
  [show abstract] [hide abstract]
  ABSTRACT: pT1 urothelial carcinoma of the bladder is a potentially aggressive cancer diathesis with heterogeneous clinical behaviors. We tested whether the combination of immunohistochemical markers could risk stratify cases of pT1 urothelial carcinoma of the bladder at radical cystectomy. p53, p21, pRB, p27, survivin and Ki-67 immunohistochemical staining was performed on representative urothelial carcinoma of the bladder specimens of 80 patients with pT1 urothelial carcinoma of the bladder treated with radical cystectomy and bilateral pelvic lymphadenectomy (median followup 61.6 months). p53 expression was altered in 25% of patients, p21 in 46%, pRB in 39%, p27 in 35%, survivin in 49% and Ki-67 in 34%. On multivariable analyses p53, p27 and Ki-67 were independently associated with urothelial carcinoma of the bladder recurrence (HR 3.66, p = 0.033; HR 3.76, p = 0.048 and HR 3.96, p = 0.021, respectively) and disease specific mortality (HR 5.25, p = 0.016; HR 3.68, p = 0.043 and HR 6.23, p = 0.009, respectively). The combination of these 3 biomarkers stratified cases into statistically significantly different risk groups for disease recurrence (p <0.001) and disease specific mortality (p <0.001). The addition of the number of altered markers increased the concordance indices of the base model that included grade, lymph node status, lymphovascular invasion and concomitant carcinoma in situ for disease recurrence and disease specific survival from 54.7% to 71.7% and from 64.3% to 77.5%, respectively. Assessment of p53, p27 and Ki-67 in urothelial carcinoma of the bladder specimens improves the prediction of recurrence-free and urothelial carcinoma of the bladder specific survival in patients with pT1 disease at radical cystectomy. These markers may help stratify the heterogeneous population of patients with pT1 disease into risk groups that can be used to guide clinical decision making regarding observation vs adjuvant therapy.
  The Journal of urology 06/2009; 182(1):78-84; discussion 84. · 4.02 Impact Factor
• Source

  Available from: Lukas C Heukamp

  Article: Multi-institutional validation of the predictive value of Ki-67 labeling index in patients with urinary bladder cancer.

  Vitaly Margulis, Yair Lotan, Pierre I Karakiewicz, Yves Fradet, Raheela Ashfaq, Umberto Capitanio, Francesco Montorsi, Patrick J Bastian, Matthew E Nielsen, Stefan C Müller, Jérôme Rigaud, Lukas C Heukamp, George Netto, Seth P Lerner, Arthur I Sagalowsky, Shahrokh F Shariat
  [show abstract] [hide abstract]
  ABSTRACT: Several small single-center studies have reported a prognostic role for Ki-67 labeling index in advanced urothelial carcinoma of the urinary bladder. To investigate whether Ki-67 was a useful biomarker of oncological outcome after radical cystectomy for urothelial carcinoma, we assessed its expression in tumor tissue from 713 patients treated with radical cystectomy and bilateral lymphadenectomy at six centers. A high Ki-67 labeling index was independently associated with established features of aggressive urothelial carcinoma, disease recurrence, and cancer-specific survival. Addition of Ki-67 labeling index improved the accuracy of standard multivariate outcome prediction models, as measured by Harrell concordance index, by 2.9% for disease recurrence and 2.4% for bladder cancer-specific survival (P < .001, two-sided Mantel-Haenszel) -- a statistically and potentially clinically significant margin. In conclusion, routine assessment of Ki-67 expression status along with assessment of other established predictors of urothelial carcinoma outcome has the potential to improve identification of patients who are at increased risk for disease progression after radical cystectomy and thus may benefit from perioperative systemic chemotherapy.
  CancerSpectrum Knowledge Environment 01/2009; 101(2):114-9. · 14.07 Impact Factor
• Article: Copy number alterations in pancreatic cancer identify recurrent PAK4 amplification.

  Shuaili Chen, Theresa Auletta, Ostap Dovirak, Christina Hutter, Karen Kuntz, Samira El-ftesi, Jude Kendall, Haiyong Han, Daniel D Von Hoff, Raheela Ashfaq, Anirban Maitra, Christine A Iacobuzio-Donahue, Ralph H Hruban, Robert Lucito
  [show abstract] [hide abstract]
  ABSTRACT: Pancreatic cancer is one of the most lethal of all cancers. The median survival is six months and less than 5% of those diagnosed survive five years. Recurrent genetic deletions and amplifications in 72 pancreatic adenocarcinomas, the largest sample set analyzed to date for pancreatic cancer, were defined using comparative genomic hybridization The recurrent genetic alterations identified target a number of previously well-characterized genes, as well as regions that contain possible new oncogenes and tumor suppressor genes. We have focused on chromosome 19q13, a region frequently found amplified in pancreatic cancer and demonstrate how boundaries of common regions of mutation can be mapped and how a gene, in this case PAK4 amplified on chromosome19q13, can be functionally validated. We show that although the PAK4 gene is not activated by mutation in cell lines with gene amplification, an oncogenic form of the KRAS2 gene is present in these cells and oncogenic KRAS2 can activate PAK4. In fact in the three samples we identified with PAK4 gene amplification, the KRAS2 gene was activated and genomically amplified. The kinase activity of the PAK4 protein is significantly higher in cells with genomic amplification as compared to cells without amplification. Our study demonstrates the utility of analyzing copy number data in a large set of neoplasms to identify genes involved in cancer. We have generated a useful dataset which will be particularly useful for the pancreatic cancer community as efforts are undertaken to sequence the pancreatic cancer genome.
  Cancer biology & therapy 12/2008; 7(11):1793-802. · 2.64 Impact Factor
• Article: Spa: Comparacion de la prueba del virus del papiloma humano con la espectroscopia en combinacion con la citologia de cervix para la deteccion de neoplasia cervical de alto grado

  Claudia L. Werner, William F. Griffith, Raheela Ashfaq, Diana Gossett, Edward Wilkinson, Stephen Raab, Shabbir Bambot, David Mongin, Mark Faupel
  [show abstract] [hide abstract]
  ABSTRACT: Objetivo. En este estudio se comparo la eficacia de la citologia cervical combinada con la prueba del virus del papiloma humano (citologia + VPH) o con la espectroscopia de cervix (citologia + EC) para identificar casos de neoplasia cervical de alto grado en una poblacion de alto riesgo de mujeres remitidas para la realizacion de una colposcopia. Materiales y metodos. A cada una de las 113 pacientes se les practico una espectroscopia, citologia en capa fina, prueba del VPH, colposcopia, biopsia cuando estaba indicada su realizacion y/o legrado endocervical. Se recopilaron datos evaluables para el analisis de 102 de las pacientes. Se calculo la sensibilidad y la especificidad de ambas estrategias. Resultados. La citologia + VPH y la citologia + EC alcanzaron unas sensibilidades equivalentes (95%) para lesiones de alto grado, y con ambas estrategias se detectaron 17 de 18 lesiones de neoplasia cervical intraepitelial (CIN) de grado 2 o superior confirmadas histologicamente. La especificidad de la citologia + VPH fue solo del 27,4% en comparacion con el 65,5% de la citologia + EC (p < 0,0001). Conclusiones. El analisis espectroscopico del cervix tiene la misma sensibilidad y el doble de especificidad que la prueba del VPH cuando se realiza en combinacion con la citologia de cervix para identificar neoplasia cervical de alto grado. [black small square] (C)2008The American Society for Colposcopy and Cervical Pathology
  Journal of Lower Genital Tract Disease 10/2008; 1(3). · 1.07 Impact Factor