Marie C. Hupe

Developing proteomic biomarkers for bladder cancer: towards clinical application

Clinical use of proteomic biomarkers has the potential to substantially improve the outcomes of patients with bladder cancer. An unmet clinical need evidently exists for noninvasive biomarkers, which might enable improvements in both the diagnosis and prognosis of patients with bladder cancer, as well as improved monitoring of patients for the presence of recurrence. Urine is considered the optimal noninvasive source of proteomic biomarkers in patients with bladder cancer. Currently, a number of single-protein biomarkers have been detected in urine and tissue using a variety of proteomic techniques, each having specific conceptual considerations and technical implications. Promising preclinical data are available for several of these proteins; however, the combination of single urinary proteins into multimarker panels might better encompass the molecular heterogeneity of bladder cancer within this patient population, and prove more effective in clinical use.

An Update on Triptorelin: Current Thinking on Androgen Deprivation Therapy for Prostate Cancer

Androgen deprivation therapy (ADT) is the mainstay palliative treatment for men with locally advanced and metastatic prostate cancer, and aims to reduce testosterone to levels obtained by surgical castration. Use of gonadotropin-releasing hormone (GnRH) agonists predominates among the ADT options. The GnRH agonist, triptorelin is a first-line hormonal therapy that has demonstrated efficacy and safety in clinical trials of patients with locally advanced non-metastatic or metastatic disease. Sustained-release 1-, 3- and 6-month formulations of triptorelin, administered intramuscularly or subcutaneously, have been developed to provide improved flexibility and convenience for the patient. Head-to-head studies of GnRH agonists are lacking in the field of prostate cancer. Despite the inevitable progression to castration-resistant prostate cancer (CRPC) in most patients receiving ADT, monitoring of testosterone levels needs to improve in routine practice and physicians should not overlook the benefits of continued ADT in their patients when introducing one of the various new treatment options for CRPC. For improved survival outcomes, there remains a need to tailor ADT treatment regimens, novel hormonal agents and chemotherapy according to the individual patient with advanced prostate cancer.

Antitumor activity of sulfated hyaluronic acid fragments in pre-clinical models of bladder cancer

Tumor cell-derived hyaluronidase HYAL-1 degrades hyaluronic acid (HA) into angiogenic fragments (AGF: 10-12 disaccharides). AGF support tumor growth and progression. Urine and tissue HAase/HYAL-1 levels are sensitive markers for high-grade bladder cancer (BCa) and its metastasis. In preclinical models of BCa, we evaluated whether o-sulfated AGF (sHA-F) inhibits HAase activity and has antitumor activity. At IC50 for HAase activity inhibition (5-20 μg/ml [0.4-1.7 μM]), sHA-F significantly inhibited proliferation, motility and invasion of HYAL-1 expressing BCa cells (253J-Lung, HT1376, UMUC-3), P<0.001. sHA-F did not affect the growth of HYAL-1 non-expressing BCa (5637, RT4, T24, TCCSUP) and normal urothelial (Urotsa, SV-HUC1) cells. sHA-F treatment induced apoptosis by death receptor pathway. sHA-F downregulated transcript and/or protein levels of HA receptors (CD44, RHAMM), p-AKT, β-catenin, pβ-Catenin(S552), Snail and Twist but increased levels of pβ-Catenin(T41/S45), pGSK-3α/β(S21/S9) and E-cadherin. sHA-F also inhibited CD44/Phosphoinositide 3-kinase (PI-3K) complex formation and PI-3K activity. AGF addition or myristoylated-AKT overexpression attenuated sHA-F effects. Contrarily, HYAL-1 expression sensitized RT4 cells to sHA-F treatment. In the 253J-L and HT1376 xenograft models, sHA-F treatment significantly inhibited tumor growth (P<0.001), plausibly by inhibiting angiogenesis and HA receptor-PI-3K/AKT signaling. This study delineates that sHA-F targets tumor-associated HA-HAase system and could be potentially useful in BCa treatment.

Proteomics analysis of bladder cancer invasion: Targeting EIF3D for therapeutic intervention

Patients with advanced bladder cancer have poor outcomes, indicating a need for more efficient therapeutic approaches. This study characterizes proteomic changes underlying bladder cancer invasion aiming for the better understanding of disease pathophysiology and identification of drug targets. High resolution liquid chromatography coupled to tandem mass spectrometry analysis of tissue specimens from patients with non-muscle invasive (NMIBC, stage pTa) and muscle invasive bladder cancer (MIBC, stages pT2+) was conducted. Comparative analysis identified 144 differentially expressed proteins between analyzed groups. These included proteins previously associated with bladder cancer and also additional novel such as PGRMC1, FUCA1, BROX and PSMD12, which were further confirmed by immunohistochemistry. Pathway and interactome analysis predicted strong activation in muscle invasive bladder cancer of pathways associated with protein synthesis e.g. eIF2 and mTOR signaling. Knock-down of eukaryotic translation initiation factor 3 subunit D (EIF3D) (overexpressed in muscle invasive disease) in metastatic T24M bladder cancer cells inhibited cell proliferation, migration, and colony formation in vitro and decreased tumor growth in xenograft models. By contrast, knocking down GTP-binding protein Rheb (which is upstream of EIF3D) recapitulated the effects of EIF3D knockdown in vitro, but not in vivo. Collectively, this study represents a comprehensive analysis of NMIBC and MIBC providing a resource for future studies. The results highlight EIF3D as a potential therapeutic target.Patients with advanced bladder cancer have poor outcomes, indicating a need for more efficient therapeutic approaches. This study characterizes proteomic changes underlying bladder cancer invasion aiming for the better understanding of disease pathophysiology and identification of drug targets. High resolution liquid chromatography coupled to tandem mass spectrometry analysis of tissue specimens from patients with non-muscle invasive (NMIBC, stage pTa) and muscle invasive bladder cancer (MIBC, stages pT2+) was conducted. Comparative analysis identified 144 differentially expressed proteins between analyzed groups. These included proteins previously associated with bladder cancer and also additional novel such as PGRMC1, FUCA1, BROX and PSMD12, which were further confirmed by immunohistochemistry. Pathway and interactome analysis predicted strong activation in muscle invasive bladder cancer of pathways associated with protein synthesis e.g. eIF2 and mTOR signaling. Knock-down of eukaryotic translation initiation factor 3 subunit D (EIF3D) (overexpressed in muscle invasive disease) in metastatic T24M bladder cancer cells inhibited cell proliferation, migration, and colony formation in vitro and decreased tumor growth in xenograft models. By contrast, knocking down GTP-binding protein Rheb (which is upstream of EIF3D) recapitulated the effects of EIF3D knockdown in vitro, but not in vivo. Collectively, this study represents a comprehensive analysis of NMIBC and MIBC providing a resource for future studies. The results highlight EIF3D as a potential therapeutic target.

Minimally invasive approaches to adrenal tumors: an up-to-date summary including patient position and port placement of laparoscopic, retroperitoneoscopic, robot-assisted, and single-site adrenalectomy.

Purpose of review There are multiple minimal invasive approaches to remove the adrenal gland. The purpose of this review is to summarize the most up-to-date findings about laparoscopic, retroperitoneoscopic, robot-assisted, and single-site adrenalectomy, and to define the most common approaches to the adrenal gland. Recent findings Laparoscopic adrenalectomy is the gold standard to remove adrenal tumors. New approaches are being explored to outperform the advantages of laparoscopic adrenalectomy. Summary Retroperitoneoscopic adrenalectomy, when performed by skilled surgeons, offers an alternative to the conventional laparoscopic approach, with better outcome. The robot-assisted and single-site approaches still need further studies to fully identify their roles in adrenalectomy.

Upper Urinary Tract Cancer

Urothelial cancers are the fourth most common cancer of which more than 95 % originate from the urothelium. The majority of these tumors arise within the bladder (90–95 %) while only 5–10 % occur in the upper urinary tract, i.e., renal pelvis and ureter (upper urinary tract urothelial cell carcinomas (UUTUCCs)). Tumors of the pyelocaliceal system are more common than ureteral tumors. Two thirds (60 %) of the UUTUCCs are invasive when diagnosed and 8–13 % of all patients present with concomitant bladder cancer. UUTUCC more commonly affects patients at approximately 80 years of age with a male to female ration of 3:1 [1]. Patients with an invasive tumor (T2-4) have a significantly lower 5-year disease-specific survival rate than patients with a superficial tumor (Ta and T1), 16.8 and 70.8 %, respectively [2]. Risk factors for UUTUCC include smoking, analgesics, chronic urinary tract infection, stone disease, and chemotherapeutic agents such as cyclophosphamide. There is also an increased risk for UUTUCC with patients who have been diagnosed with primary bladder cancer (10 %). Prognostic factors include stage, grade, lymph node invasion, lymphovascular invasion, tumor necrosis, and tumor architecture (infiltrative) [3].

The BET-inhibitor PFI-1 diminishes AR/AR-V7 signaling in prostate cancer cells

ObjectiveThe bromodomain and extra-terminal (BET) family of proteins provides a scaffolding platform for the recruitment and tethering of transcription factors to acetylated chromatin, thereby modulating gene expression. In this study, we evaluated the efficacy of the BET-inhibitor PFI-1 to diminish AR/AR-V7 signaling and proliferation in castration-resistant prostate cancer cells.MethodsProstate-specific antigen and androgen receptor (AR) protein were quantified by means of two commercial ELISAs. Transactivation of the AR, AR-V7 and Q641X was determined by reporter gene assays. Cell proliferation was measured using a colorimetric MTT-assay.ResultsPFI-1 dose-dependently inhibited transactivation of full-length AR (non- mutated, i.e., wild-type or point-mutated/promiscuous forms) without affecting their cellular protein levels. Moreover, PFI-1 was active against C-terminally truncated constitutively active ARs like AR-V7 and Q641X. Prostate cancer cells exhibiting a transcriptionally active AR-signaling complex (LNCaP, 22Rv1) were more susceptible to the growth-inhibitory effects than the AR-negative PC-3 cells.ConclusionThe quinazolinone PFI-1 is a highly efficient inhibitor of AR-signaling-competent prostate cancer cells in vitro. PFI-1 could serve as a lead compound for the development of new therapeutics able to block AR/AR-V7 signaling in advanced prostate cancer.

Diarrhea and flatulence are major bowel disorders after radical cystectomy: Results from a cross-sectional study in bladder cancer patients

OBJECTIVES We had previously demonstrated changes in defecation after radical cystectomy (RC). Reports addressing long-term bowel disorders following RC are rare. This cross-sectional study evaluates long-term bowel issues in a large cohort. MATERIAL AND METHODS A questionnaire assessing changes in bowel function (diarrhea, constipation, urge to defecate, sensation of incomplete defecation, and flatulence) and its effect on quality of life was developed based on the gastrointestinal quality of life index and distributed in collaboration with the German bladder cancer support group. There were 431 evaluable questionnaires. For the analyses, we focused on patients that had the RC>1 year ago (n = 324). RESULTS Current bowel problems were reported by 42.6% of patients. The most frequent bowel problems were flatulence (48.8%), diarrhea (29.6%), and sensation of incomplete defecation (22.5%). In cases of bowel problems, 39.7% and 59.8% of the patients indicated life restriction and dissatisfaction, respectively. Prevalence of diarrhea and flatulence were significantly higher>12 (vs. ≤12) months following RC. Both symptoms significantly correlated with younger age at RC, life restriction, lower quality of life, lower health state, and lower energy level. Additionally, diarrhea significantly correlated with pouches as urinary diversion (vs. ileal conduit or ureterocutaneostomy) and higher dissatisfaction level. CONCLUSIONS To our knowledge this is the largest cohort evaluating long-term bowel symptoms after RC. Diarrhea is a prominent symptom after RC with a high impact on daily life that leads to dissatisfaction. A better understanding of long-term bowel symptoms could be translated into optimized surgical procedures, postoperative medication/nutrition, and patient education.

Extended Techniques of Minimally Invasive Surgery in the Retroperitoneum: Practice Pattern in German Urology Departments

Introduction: Laparoendoscopic single-site surgery (LESS), robot-assisted (RA), and retroperitoneoscopic (R) surgery expand the armamentarium of minimally invasive surgery (MIS). As information on the use of these surgical approaches in daily routine is limited, we conducted a survey among German urologists. Materials and Methods: In 2017, all urology departments in Germany received a questionnaire evaluating practice patterns of MIS in the retroperitoneum. Chi-Square test was performed for statistical analyses. The response rate was 51.1% (162/311) including 23 universities. Results: R adrenalectomy and (partial) nephrectomy are performed by 32.7–40.1% of all departments. Transperitoneal LESS adrenalectomy and nephrectomy are performed by 8.6–11.7%. Retroperitoneal RA adrenalectomy and (partial) nephrectomy are performed by 6.2–13.0%. There was no difference in the R and LESS approach between (non)-university departments. Retroperitoneal RA access is more frequently used in university hospitals (all p < 0.01). If performed, mean counts within the last 12 months were < 5 for R, LESS, and RA adrenalectomy; and < 20 for R, LESS, and RA (partial) nephrectomy. Conclusion: Our survey provides a detailed insight into MIS in the retroperitoneum in German urology departments. Numbers of adrenalectomies as well as R, transperitoneal LESS and retroperitoneal RA procedures are low. Retroperitoneal RA surgery is more common in universities. Comprehensively, our survey proves that these approaches are not standard approaches yet.

MP39-15 SDCT2 AS A FUNCTIONAL BIOMARKER OF RENAL CELL CARCINOMA

patients were stained for iron and TfR1 protein using Prussian Blue and immunohistochemistry, respectively. 178 tissue cores from 14 different body sites of non-cancer patients were included as controls. Staining was scored by a clinically blinded genitourinary pathologist based on the product of intensity and tissue percentage (Z-score), and tested for association with clinicopathologic features and survival using a MannWhitney U test, Kruskal Wallis test and Cox regression model. RESULTS: Renal epithelium from non-cancer patients had low iron content (mean Z-score,MZS1⁄4 0.1), but by far the highest TfR1 levels of any tissue site in the body (MZS1⁄4 153). Compared to non-cancer patients, iron content in RCC patients increased mildly (4-fold) in benign renal epithelium (MZS1⁄4 0.6) and dramatically (>100 fold) in primary tumors (MZS1⁄4 21, p< 0.001). Higher tumor iron content was accompanied by moderate TfR1 downregulation (MZS1⁄4 21, p< 0.001) and associated with clear cell and papillary histologies, male gender and tobacco usage (p< 0.05 each). Opposite to changes observed with tumorigenesis, iron and TfR1 levels decreased and increased, respectively, with progression in tumor size, grade, pT stage andmetastatic stage (all p< 0.05). Iron loss andTfR1upregulationweremost apparent inmetastatic lesions (MZS1⁄45 and 111, respectively) and each associated with patient anemia and worse RCC-specific survival (all p< 0.05). CONCLUSIONS: Benign renal epithelium has uniquely high levels of the iron import protein, TfR1, potentially priming these cells for dysregulated iron uptake and large intracellular iron increases (>100 fold) during tumorigenesis. Reduction in iron content during RCC progression to metastasis, despite TfR1 increases, may reflect lower systemically available iron in advanced RCC patients and raises the possibility that these cancers might have increased susceptibility to iron deprivation as a novel therapeutic strategy.