Muscle Invasion Research Articles

Multi-Sequence and Multi-Regional MRI-Based Radiomics Nomogram for the Preoperative Assessment of Muscle Invasion in Bladder Cancer.

Whether bladder cancer (BCa) invades muscle is a determinant of management. However, the accuracy of preoperative diagnosis of muscle invasion is not satisfactory. To investigate the value of multi-sequence and multi-regional magnetic resonance imaging (MRI)-based radiomics nomogram for assessing muscle invasion of BCa. Retrospective. 342 BCa patients, divided into a training set (239 cases), a validation set (68 cases), and a test set (35 cases). 3.0 T/T2 -weighted image, diffusion-weighted imaging, and dynamic contrast-enhanced imaging. Patients were divided into muscle-invasive (79 cases) and non-muscle-invasive (263 cases). Two radiologists delineated the whole tumor, tumor body, and muscle layer of BCa, respectively, and extracted radiomic features. Recursive feature elimination, Pearson correlation coefficient, logistic regression, least absolute shrinkage and selection operator (Lasso) regression analysis, and 5-fold cross-validation were used to screen features and build a radiomics model. The clinical data were collected to construct a clinical model and a radiomics-clinical nomogram. 23,688 features were extracted. After screening, the radiomics scoring model was constructed using nine radiomics features with area under curve (AUC) values of 0.933, 0.913, and 0.931 in the training, validation, and test sets, respectively. The clinical model was constructed using five clinical independent risk factors; the AUC values in the training, validation, and test set were 0.876, 0.859, and 0.824, respectively. After logistic regression analysis, the AUC values of the radiomics-clinical nomogram were made up of four clinical independent risk factors and radiomics scores were 0.955, 0.922, and 0.935 for the training, validation, and test sets, respectively. The DeLong test between clinical model and radiomics-clinical nomogram shows P < 0.001. Multi-sequence and multi-regional MRI-based radiomics models could effectively assess the state of BCa muscular invasion. The radiomics-clinical nomogram is superior to clinical model for assessing BCa muscular invasion. 4 TECHNICAL EFFICACY: Stage 2.

High Pretreatment Serum PD-L1 Levels Are Associated with Muscle Invasion and Shorter Survival in Upper Tract Urothelial Carcinoma.

Simple SummaryThis study aimed to assess the prognostic relevance of soluble serum PD-L1 in upper tract urothelial carcinoma (UTUC) patients who underwent surgical or systemic (chemo- or immune checkpoint inhibitor) therapy. We found that high preoperative sPD-L1 levels were correlated with higher pathological tumor stage, grade and the presence of metastasis. In addition, higher pretreatment serum PD-L1 concentrations were associated with shorter survival in both surgically and chemotherapy-treated UTUC patients. We detected a characteristic increase in serum PD-L1 levels in UTUC patients after 3 months of anti-PD-L1 therapy. Based on these results sPD-L1 is a promising prognostic biomarker in UTUC.Programmed death ligand-1 (PD-L1) is an immune checkpoint molecule and a widely used therapeutic target in urothelial cancer. Its circulating, soluble levels (sPD-L1) were recently suggested to be associated with the presence and prognosis of various malignancies but have not yet been investigated in upper tract urothelial carcinoma (UTUC). In this study, we assessed sPD-L1 levels in 97 prospectively collected serum samples from 61 UTUC patients who underwent radical nephroureterectomy (RNU), chemotherapy (CTX), or immune checkpoint inhibitor (ICI) therapy. In addition to pretreatment samples, postoperative and on-treatment sPD-L1 levels were determined in some patients by using ELISA. In the RNU group, elevated preoperative sPD-L1 was associated with a higher tumor grade (p = 0.019), stage (p < 0.001) and the presence of metastasis (p = 0.002). High sPD-L1 levels were significantly associated with worse survival in both the RNU and CTX cohorts. sPD-L1 levels were significantly elevated in postoperative samples (p = 0.011), while they remained unchanged during CTX. Interestingly, ICI treatment caused a strong, 25-fold increase in sPD-L1 (p < 0.001). Our results suggest that elevated preoperative sPD-L1 level is a predictor of higher pathological tumor stage and worse survival in UTUC, which therefore may help to optimize therapeutic decision-making. The observed characteristic sPD-L1 flare during immune checkpoint inhibitor therapy may have clinical significance.

Prospective assessment of VI-RADS score in multiparametric MRI in bladder cancer: accuracy and the factors affecting the results.

PURPOSE We aimed to investigate the accuracy of Vesical Imaging - Reporting and Data System (VI-RADS) in the detection of muscle-invasive bladder cancer (MIBC) and to determine which factors affect the results of this scoring system. METHODS A prospective data analysis of 80 patients who were detected to have bladder tumor was performed between March 2019 and October 2020. VI-RADS scoring was used to determine the probability of muscle invasion. The scores were compared with pathological results to evaluate the accuracy of the VI-RADS scoring system. Interobserver agreement was assessed by VI-RADS scoring of 20 randomly chosen patients by a different experienced radiologist. RESULTS Using the VI-RADS scoring system, the sensitivity, specificity, positive predictive value, and negative predictive value of multiparametric magnetic resonance imaging (mpMRI) were 87.5%, 87.5%, 63.6%, and 96.6%, respectively. The interobserver agreement expressed as the interclass correlation coefficient (ICC) was 0.72 (95% CI: 0.44-0.84, P < .001). In addition, the flat appearance of the tumor was an important factor affecting the accuracy of the VI-RADS score (odds ratio: 5.3 [95% CI: 1.1-27.0] and relative risk: 1.87 [95% CI: 1.24-2.82]). CONCLUSION The mpMRI, used in conjunction with VI-RADS, has proven to be an effective imaging method for detecting muscle invasion in cases of bladder cancer. VI-RADS scoring system can distinguish whether there is a muscle-invasive and non-muscle invasive bladder cancer with acceptable accuracy. In addition, the flat appearance of the tumor is an important entity that can affect the accuracy of the VI-RADS scoring system.

Expression of Vitamin D Receptor (VDR) in Urinary Bladder Carcinoma: Immunohistochemical and Histopathological Study

Background: Bladder cancer is the most common malignancy of the urinary tract. Calcitriol [1,25 (OH)2vitamin D3] has anticancer effects mediated through binding to vitamin D receptor (VDR). The expression of VDR is present in many normal and cancer tissues. But, there is little information about its expression in urinary bladder carcinoma. This study aimed to analyze VDR immunohistochemical expression in 74 Egyptian patients with urinary bladder carcinoma and to evaluate its association with different clinicopathological parameters. Methods: Sections from formalin-fixed, paraffin-embedded tumor blocks were stained immunohistochemically using monoclonal anti-VDR antibody. VDR protein expression as well as its immunostaining patterns were recorded and scored separately in each case using semi-quantitative immunoreactive score. Results: VDR was consistently expressed in the included histologically normal urothelium while tumor cells showed variable degrees of expression. Cytoplasmic/membranous VDR expression was common among the studied cases especially those with urothelial morphology (p = 0.076). While, the mean nuclear VDR was significantly (p = 0.007) higher in non-urothelial tumors. Nuclear VDR was significantly associated with muscle invasion (p = 0.000) and tumor stage (p = 0.001) in urothelial carcinoma. It was also statistically related to tumor grade, stage and muscle invasion in non-urothelial tumors (p = 0.002, 0.003 and 0.012, respectively). Conclusion: there was a significant relation between nuclear VDR expression and prognostic markers suggesting its decrease as an indicator of a poorer prognosis. Vitamin D supplementation may represent a new treatment option for patients with bladder cancer.

Expression of GLUT4 and FAP in urothelial bladder carcinoma: correlation with angiogenesis and clinicopathological characteristics

BackgroundUrothelial carcinoma (UC) is the most common type of bladder cancer. Glucose transporter 4 (GLUT4) is one of glucose transporter proteins’ family which facilitates glucose transport inside the cells. It was found to be overexpressed in several malignant tumors. Cancer-associated fibroblasts (CAFs) are heterogeneous stromal cells located adjacent to cancer cells and are considered one of the most important tumor stromal cells. They have been associated with enhancing tumor growth and invasion. GLUT4 expression in malignant epithelial cells and fibroblast activation protein (FAP) expression in CAFs of UC in relation to angiogenesis and clinicopathological characteristics are studied in this work.Materials and methodsThe study was carried out on 72 paraffin blocks of UC (27 radical cystectomies and 45 transurethral resections). Immunohistochemical staining was performed with GLUT4, FAP, and CD34 antibodies. Expression of GLUT4 and FAP was classified according to the staining intensities and percentages into low and high groups. CD34-stained microvessels’ mean count in five microscopic fields (×200) was taken as the microvessel density (MVD).ResultsGLUT4 overexpression was detected in 32 UC. It was significantly associated with high-grade tumors, advanced primary tumor (pT) stage, lymphovascular invasion (LVI), and regional lymph node invasion. High FAP expression was appreciated in 27 UC and was significantly linked to LVI and advanced TNM staging. Intratumor MVD significantly increased in UC with muscle invasion, LVI, and regional lymph node and/or distant metastasis. A significant positive correlation between GLUT4, FAP expression, and MVD was found.ConclusionGLUT4 and FAP expression was significantly associated with increased intratumor MVD and adverse clinicopathological factors.

Clinical outcomes of gastroduodenal neuroendocrine tumors according to their WHO grade: A single-institutional retrospective analysis.

The management of gastroduodenal neuroendocrine tumor (NET) has been controversial between radical surgical resection and local excision including endoscopic resection. A gastroduodenal NET grade (G), measured by their mitotic rate and Ki67 proliferation index, is important to predict prognosis. In this study, we aimed to compare the clinical outcomes of gastroduodenal NET according to grades in order to identify poor prognostic factors of gastroduodenal NETs. Fifty-four gastroduodenal NETs diagnosed between December 2008 and December 2020 in a tertiary referral hospital were retrospectively reviewed. The clinical outcomes of gastroduodenal NETs, according to tumor grades and factors associated with NET G2-3, were analyzed. A total of 52 gastroduodenal NET patients was enrolled. The mean follow-up period was 56.2 ± 40.1 months. The mean size of gastric and duodenal NET was 7.9 ± 11.0 mm and 9.8 ± 7.6 mm, respectively. During the study period, 72.7% (16/22) of gastric NETs and 83.3% (25/30) of duodenal NETS were G1. All G1 gastroduodenal NETs showed no lymph node or distant metastasis during the study periods. All G3 gastroduodenal NETs showed metastasis (one lymph node metastasis and 3 hepatic metastases). Among metastatic NETs, the smallest tumor size was a 13 mm gastric G3 NET. Factors associated with G2-3 NETs were larger tumor size, mucosal ulceration, proper muscle or deeper invasion, and lymphovascular invasion. A small-sized gastroduodenal NET confined to submucosa without surface ulceration may be suitable for endoscopic resection. After local resection of a gastroduodenal NET (G1) without lymphovascular and muscle proper invasion, follow-up examination without radical surgical resection can be recommended. G3 NETs may be treated by radical surgical resection, regardless of tumor size.

Application of Vesical Imaging-Reporting and Data System in Evaluation of Urinary Bladder Cancer Using Multiparametric Magnetic Resonance Imaging: A Hospital-Based Cross-Sectional Study.

Background Multiparametric magnetic resonance imaging (mp-MRI) of urinary bladder (UB) is a novel imaging to predict detrusor muscle invasion in Bladder cancer (BC). The Vesical Imaging-Reporting and Data System (VI-RADS) was introduced in 2018 to standardize the reporting of BC with mp-MRI and to diagnose muscle invasion. This study was performed to evaluate the role of mp-MRI using VI-RADS to predict muscle invasive BC. Methods This prospective study was carried from June 2020 to May 2021 in a tertiary care institute. Thirty-six patients with untreated BC underwent mp-MRI followed by transuretheral resection of the tumor (TURBT). Mp-MRI findings were evaluated by two radiologists and BC was categorized according to VI-RADS scoring system. Resected tumors along with separate biopsy from the base were reported by two pathologists. Histopathological findings were compared with VI-RADS score and the performance of VI-RADS for determining detrusor muscle invasion was analyzed. Results VI-RADS scores of 4 and 5 were assigned to 9 (25%) and 15 (41.7%) cases, respectively, while 4 (13.3%) cases had VI-RADS score 3 on mp-MRI. VI-RADS 1 and 2 lesions were observed in six (16.7%) and two (5.5%) cases, respectively. On histopathology, 23 cases (63.9%) had muscle-invasive cancer and 13 cases (36.1%) had non-muscle-invasive cancer. The sensitivity and diagnostic accuracy of mp-MRI in predicting muscle invasive BC was 95.6 and 80.6%, respectively. Conclusion Mp-MRI has high sensitivity and diagnostic accuracy in predicting muscle invasive BC and should be advocated for evaluation of BC prior to surgery.

Endoscopic Management of Malignant Tumors of Paranasal Sinus and Nasal Cavity: An Institutional Experience.

Malignant tumors of sinonasal region are rare and affect less than 1 in 100,000 people per year. They are histologically diverse group and potentially pose significant management problems due to their proximity to the orbit and intracranial cavity. Although squamous cell carcinoma (SCC) is most common malignant tumor of paranasal cavity, tumors like adenocarcinoma, olfactory neuroblastoma, malignant melanoma, adenoid cystic carcinoma, sarcomas, haemoproliferative tumors, e.g. lymphoma may also occur. Retrospectively study was done in a tertiary care institute from January 2008 to December 2018 in India. Inclusion criteria-all biopsy proven PNS malignancy patients operated by endoscopic approach, irrespective of age and gender. Exclusion criteria- patients diagnosed with nasopharyngeal carcinoma, skin involvement, gross orbital involvement (muscle invasion), metastasis, operated by open approaches. 46 patients who underwent endoscopic tumor removal were reviewed. 36 (78.2%) were males and 10 (4.6%) females. Most common tumor in our study was adenoid cystic carcinoma. Recurrence was seen in 6 patients. Palliative therapy was given to all patients with recurrence. Management of malignant PNS tumor involving anterior skull base is multidisciplinary. R0 resection should be main goal in all malignant PNS malignancy. Tumors invading unresectable areas like cavernous sinus, brain parenchyma, carotids can be left in these places followed by palliation except in cases of squamous cell carcinoma. If R0 cannot be achieved surgically in SCC than patient should be considered inoperable and send for palliation. T1-T3 PNS malignant tumors can be managed by endoscopic approach followed by adjuvant therapy after a period of 6weeks. Endoscopic excision should be converted to endoscopic assisted open approach in case of T4 tumors. We have tried to give a management protocol for management of malignant PNS tumors. Level of evidence: II.

Gasdermin D: A potential mediator and prognostic marker of bladder cancer.

Background: Bladder cancer is considered one of the commonest widespread cancers, its presentation ranges from non-muscle invasive form to being muscle-invasive. The gasdermin family of proteins consists of six proteins. Members of gasdermin family are involved in pyroptosis; which is considered as type of inflammatory apoptosis via participation of gasdermin D and inflammatory caspases. Purpose: The goal of this research was to look into the potential involvement of gasdermin D in pathogenesis of bladder cancer, In addition, to investigate its potential role as a prognostic marker of bladder cancer. Methods: Gasdermin D gene and protein expression was examined in fresh frozen 80 bladder cancer specimens (30 NMIBC, and 50 MIBC) and the matching 80 control tissue samples utilizing real-time polymerase chain reaction and western blotting. Furthermore, the immunoreactivity of gasdermin D protein was also detected by immunohistochemistry. Results: Gasdermin D gene and protein expression showed a highly significant difference between the control and the two bladder cancer groups (p < 0.001), as demonstrated by real-time PCR, western blotting and immunohistochemistry. Cox proportional hazards regression models showed that lower gasdermin D gene expression in cancer patients (≤1.58-fold), and younger age (≤53 years) were linked with a higher risk of local tumor recurrence. Moreover, higher gasdermin D gene expression (>2.18-fold), and lymph nodes’ involvement were associated with an increased mortality. Conclusion: Gasdermin D is involved in the pathogenesis of bladder cancer and muscle invasion, in addition, tissue gasdermin D expression may be used as useful tool to predict local tumor recurrence.

Role of multiparametric magnetic resonance imaging in the diagnosis and staging of urinary bladder cancer.

ObjectivesTo assess the role of multiparametric magnetic resonance imaging (mp-MRI) in the diagnosis and staging of urinary bladder cancer (BC).Materials and methodsFifty patients diagnosed with bladder masses underwent mp-MRI study. The results of 3 image sets were analyzed and compared with the histopathological results as a reference standard: T2-weighted image (T2WI) plus dynamic contrast-enhanced (DCE), T2WI plus diffusion-weighted images (DWI), and mp-MRI, including T2WI plus DWI and DCE. The diagnostic accuracy of mp-MRI was evaluated using receiver operating characteristic curve analysis.ResultsThe accuracy of T2WI plus DCE for detecting muscle invasion of BC was 79.5% with a fair agreement with histopathological examination (κ = 0.59); this percentage increased up to 88.6% using T2WI plus DWI, with good agreement with histopathological examination (κ = 0.74), whereas mp-MRI had the highest overall accuracy (95.4%) and excellent agreement with histopathological data (κ = 0.83). Multiparametric MRI can differentiate between low- and high-grade bladder tumors with a high sensitivity and specificity of 93.3% and 98.3%, respectively.ConclusionsMultiparametric MRI is an acceptable method for the preoperative detection and accurate staging of BC, with reasonable accuracy in differentiating between low- and high-grade BC.

The role of radiomics with machine learning in the prediction of muscle-invasive bladder cancer: A mini review.

Bladder cancer is a common malignant tumor in the urinary system. Depending on whether bladder cancer invades muscle tissue, it is classified into non-muscle-invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC). It is crucial to accurately diagnose the muscle invasion of bladder cancer for its clinical management. Although imaging modalities such as CT and multiparametric MRI play an important role in this regard, radiomics has shown great potential with the development and innovation of precision medicine. It features outstanding advantages such as non-invasive and high efficiency, and takes on important significance in tumor assessment and laor liberation. In this article, we provide an overview of radiomics in the prediction of muscle-invasive bladder cancer and reflect on its future trends and challenges.

Diagnostic Value of the Vesical Imaging-Reporting and Data System in Bladder Urothelial Carcinoma with Variant Histology

The value of the Vesicle Imaging-Reporting and Data System (VI-RADS) in the diagnosis of muscle-invasive bladder cancer (MIBC) for urothelial carcinoma with variant histology (VUC) remains unknown. We retrospectively evaluated 360 consecutive patients with bladder cancer (255 pure urothelial carcinoma [PUC] and 69 VUC) who underwent multiparametric magnetic resonance imaging between 2011 and 2019. VI-RADS scores assigned by four readers were significantly higher for the VUC group than for the PUC group (p < 0.05). In the cohort of 122 pair-matched patients, there was no significant difference in VI-RADS score distribution between the PUC and VUC groups for all readers (p > 0.05). The area under the receiver operating characteristic curve for MIBC diagnosis via overall VI-RADS score was 0.93–0.94 for PUC and 0.89–0.92 for VUC, with no significant difference between the PUC and VUC groups (p = 0.32–0.60). These data suggests that VI-RADS scores achieved high diagnostic performance for detection of muscle invasion in both PUC and VUC. Patient summaryThe Vesical Imaging-Reporting and Data System (VI-RADS) is a standardized system for reporting on detection of muscle-invasive bladder cancer via magnetic resonance imaging (MRI) scans. Our study shows that VI-RADS is also highly accurate for diagnosis for different variants of muscle-invasive bladder cancer, with good inter-reader agreement.

Proposal for a New Vesical Imaging-Reporting and Data System (VI-RADS)-Based Algorithm for the Management of Bladder Cancer: A Paradigm Shift From the Current Transurethral Resection of Bladder Tumor (TURBT)-Dependent Practice.

Transurethral resection of bladder tumor (TURBT) is the essential first step in the current algorithm for the management of bladder cancer (BC). However, despite its necessity and significance, TURBT has several limitations, including cost, hospitalization, anesthesia, potential complications such as bladder perforation, and delay to radical cystectomy. The Vesical Imaging Reporting and Data System (VI-RADS) was developed to standardize the reporting of multiparametric magnetic resonance imaging for BC, and its diagnostic accuracy to predict muscle invasion has been validated. Given the high sensitivity of VI-RADS ≥ 3 and high specificity of VI-RADS ≥ 4 as clinically relevant cutoff values, we herein propose a new VI-RADS-based algorithm for the management of BC. Using this algorithm, patients with VI-RADS ≤ 2 may not need to undergo sampling of the detrusor muscle nor second TURBT even if there is no muscle in the initial TURBT specimen, whereas patients with VI-RADS ≥ 4 may skip conventional TURBT aimed at pathologic confirmation of muscle invasion and immediately undergo radical cystectomy. Our newly proposed algorithm enables the avoidance of unnecessary deep resection or second TURBT as well as delay to radical cystectomy. The VI-RADS-based algorithm enables a paradigm shift from the current TURBT-dependent practice in the management of BC.

ATF4 promotes brain vascular smooth muscle cells proliferation, invasion and migration by targeting miR-552-SKI axis

BackgroundStudies have indicated vascular smooth muscle cells (VSMCs) played a crucial role in atherosclerosis and microRNAs (miRNAs) played key roles in biological functions of VSMCs. Whereas, the potential function and mechanism of miR-552 in VSMCs remains unclear. Our aim was to explore the role of miR-552 on VSMCs and underlying mechanism.Material/MethodsMTT assay and transwell assay were used to measure the proliferation, invasion, and migration of human brain VSMCs (HBVSMCs) and mice VSMCs (mVSMCs), respectively. Bioinformatics tools and luciferase assay were adopted to verify the association between miR-552 and SKI. Rescue experiments were employed to assess the interaction of miR-552 and SKI in modulating biological functions in HBVSMCs and mVSMCs. The expression level of transcription factors (TFs)was measured via qRT-PCR assay. The effect of ATF4 on miR-552 and SKI expression was tested by qRT-PCR or western blot assay. Finally, chromatin immunoprecipitation (ChIP) assay and JASPAR databases were used to analyze the regulatory linkage between ATF4 and miR-552.ResultsWe found that miR-552 was upregulated in HBVSMCs treated with PDGF-bb and miR-552 overexpression could promote proliferation, invasion, and migration of HBVSMCs and mVSMCs, whereas, miR-552 knockdown had the opposite impact. In addition, we also found that SKI was a direct target of miR-552, which reversed miR-552-mediated proliferation, invasion, and migration in HBVSMCs and mVSMCs. Furthermore, we also discovered that miR-552 overexpression promoted the effects of ATF4 elevation on proliferation, migration and invasion of HBVSMCs and mVSMCs, but, miR-552 decline had the opposite impact.ConclusionsATF4-miR-552-SKI axis played critical roles in the proliferation and migration of HBVSMCs and mVSMCs, which were closely involved in atherosclerosis (AS). Therefore, our findings might offer a novel therapeutic target for AS.

The use of MRI in urothelial carcinoma.

To conduct a nonsystematic review of the existing literature to investigate the role of Magnetic Resonance Imaging (MRI) in urothelial carcinomas (UCs). Imaging is becoming progressively more crucial in local and distant staging of UCs, especially in cases of bladder cancer (BCa). The primary objective of imaging in patients with BCa is to detect lesions and distinguish T1 from T2 stages, since the treatment varies significantly. The applicability of MRI in the management of UCs has been investigated, particularly focusing on the new evidence on multiparametric MRI (mpMRI) of the bladder and Vesical Imaging-Reporting And Data System score for the description of BCa and discussing the possible utility of MRI for upper tract urothelial carcinomas . Imaging modalities, in particular CT and MRI, are essential tools for the local and distant staging of UCs. MpMRI of the bladder and VI-RADS score accurately define the risk of muscle invasiveness, promoting tailored therapeutic planning. Moreover, mpMRI has also been included in patients' follow-up and in the assessment of response to systematic therapy. MRI utility and possible application in upper tract urothelial carcinomas cases are yet to be discovered.

The Origin and Evolution of Bladder Cancer Stem Cells.

Bladder cancer is the most common malignant tumor of the urinary system. Bladder cancer stem cells (BCSCs) play key roles in tumor initiation, metastasis, relapse and drug-resistance. Investigation of BCSCs is of great value. On the basis of a review of normal bladder stem cells and universal cancer stem cells (CSCs), we summarize the origin of BCSCs, isolation and identification of CSCs from bladder cancer, signaling pathway of BCSCs, BCSCs targeted therapy, and relationship of BCSCs with non-muscle invasiveness and muscle invasiveness. This review aims to provide better elucidation about BCSCs, and provide constructive data for classification, prognosis, treatment and early intervention of bladder cancer.

Fibroblast activation protein (FAP) expression in CK5/6 expressed (Basal subtype) & CK20 expressed (Luminal subtype) urothelial bladder carcinoma: an immunohistochemical study

ABSTRACT Urinary bladder cancer incidence varies all over the world. Egypt displays high incidence rates. Molecular subtyping helps risk stratification and personalized treatment. Cancer-associated fibroblasts (CAFs) in the tumor microenvironment may provoke tumor-promotion or tumor suppression. Fibroblast activation protein (FAP) is a marker of CAFs, suggested to accelerate tumor progression in various cancers. In urothelial carcinoma, investigations regarding impact of FAP expression on prognosis are needed. This work aims to study impact of FAP expression in urothelial carcinoma and find its relation to CK 5/6 (basal) expressed and CK 20 (luminal) expressed immunohistochemical markers. This retrospective study included 70 urothelial carcinoma specimens. Immunohistochemistry was performed and results were analyzed. FAP was expressed in 67.1% of cases and showed significant association with advanced tumor stage, muscle invasion, mitoses in tumor cells and stratified groups; as 73.9% of FAP positive cases were of Ck5/6+/Ck20- (basal subtype). All studied parameters did not show significant association with patient’s overall survival. In conclusion, FAP could have a role in modulating tumor microenvironment and promoting tumor invasion. FAP is correlated with basal subtype of urothelial carcinoma, which may be an indicator of tumor aggressiveness. FAP antagonists may be helpful in preventing tumor progression.

Intratumoral Switch of Molecular Phenotype and Overall Survival in Muscle Invasive Bladder Cancer.

Simple SummaryThe Cancer Genome Atlas (TCGA) and more recent genome profiling recently revealed major intrinsic molecular subtypes in urothelial carcinoma (UC). Here we propose a fast and standardized immunophenotypical classification score (Piescore) that may discriminate between luminal, basal, or neu-like UC as a surrogate of molecular profile, and we describe, for the first time, an intratumoral phenotypical switch in tissue protein expression, from non-muscle to muscle-invasive progression. Our data show that a change from a luminal to a neu-like phenotype could worsen overall survival compared with a transition to a basal phenotype.In recent years, immunohistochemical protein expression was studied as a surrogate to the molecular classification of bladder cancer, although no tissue biomarkers are available for clinical use to predict survival or the response to neoadjuvant chemotherapy (CT) in UC, as the literature produced conflicting results. This retrospective study included TURB specimens harboring foci of HG pT2 muscle-invasive bladder carcinoma (MIBC) from 251 patients who subsequently underwent radical cystectomy. We performed immunohistochemical analysis on tumor samples, for relevant gene-expression-based markers for basal type (CD44, CK5/6) and luminal type (CK20 and pPARγ). Piescore, investigated in both non-muscle-invasive (NMI) and muscle-invasive (MI) components of the tumor, divided basal and luminal UC-types when at least three of the four markers were consistent with a specific phenotype, mixed types if one/two luminal and basal markers were present simultaneously, and neu-like types when all four markers investigated were negative. Eighteen selected cases were also investigated with RT-PCR to validate, and to increase the specificity of, the immunohistochemical results. We observe an immunophenotypical difference in the NMI and MI components in 96/251 UC patients (38.25%): half of tumors (44/96 cases) have a transition to basal, 36.46% (35/96 cases) to neu-like, 12.5% (12/96 cases) to mixed, and 5.2% (5/96 cases) to luminal phenotypes. Mixed tumors in the NMI component are more likely to change phenotype than other groups, particularly compared with basal tumors, which demonstrate greater stability (only 8/96 cases, p < 0.00001). The transition of luminal tumors to basal display a better OS compared with the transition toward neu-like tumors (p = 0.027). Overall, the phenotypical switch does not affect lymphovascular invasion, pT, DFS, or OS compared with non-switched cases. In the MI component, the presence of CD44 expression, irrespective of score-related phenotype, shows a protective effect in papillary-type UC (OS p = 0.008, HR 0.453, PFS p = 0.07, HR 0.599), and in UC naïve for CT (p = 0.0479). Piescore immunophenotyping reveals an intratumoral phenotypical transition between the NMI and MI components of the same tumor. The molecular change is a common event in the mixed and luminal categories, but not in basal tumors, which show better phenotypical stability. This phenomenon could partially explain the sensitivity of a subset of luminal UC to chemotherapy: good responders could be “non-real” luminal UC, which acquire nasal markers, such as CD44.

The carbonic anhydrase inhibitor acetazolamide inhibits urinary bladder cancers via suppression of β-catenin signaling.

Carbonic anhydrases (CAs) play an important role in maintaining pH homeostasis. We previously demonstrated that overexpression of CA2 was associated with invasion and progression of urothelial carcinoma (UC) in humans. The purpose of the present study was to evaluate the effects of the CA inhibitor acetazolamide (Ace) on N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine (BBN)‐induced bladder carcinogenesis in mice and explore the function of CA2 in muscle invasion by UC. Male mice were treated with 0.025% (experiment 1) or 0.05% BBN (experiment 2) in their drinking water for 10 weeks, then treated with cisplatin (Cis), Ace, or Cis plus Ace for 12 weeks. In experiment 1, the overall incidence of BBN‐induced UCs was significantly decreased in the BBN→Ace and BBN→Cis+Ace groups. In experiment 2, the overall incidence of BBN‐induced UCs was significantly decreased in the BBN→Cis+Ace group, and the incidence of muscle invasive UC was significantly decreased in both the BBN→Ace and the BBN→Cis+Ace groups. We also show that overexpression of CA2 by human UC cells T24 and UMUC3 significantly increased their migration and invasion capabilities, and that Ace significantly inhibited migration and invasion by CA2‐overexpressing T24 and UMUC3 cells. These data demonstrate a functional association of CA2 with UC development and progression, confirming the association of CA2 with UC that we had shown previously by immunohistochemical analysis of human UC specimens and proteome analysis of BBN‐induced UC in rats. Our finding that inhibition of CA2 inhibits UC development and muscle invasion also directly confirms that CA2 is a potential therapeutic target for bladder cancers.

Implications for pelvic lymph node irradiation in definitive chemoradiotherapy of node negative muscle invasive bladder cancer based on predictive factors of clinicopathologic discrepancy.

To identify pre-surgical imaging predictive value and factors associated with the clinicopathologic discrepancy for implication of definitive pelvic radiotherapy in clinically node-negative bladder cancer. The documented data of bladder cancer patients who underwent radical cystectomy plus pelvic lymphadenectomy were collected retrospectively. Patients' characteristics, last imaging, pathology reports, disease-specific survival and overall survival were retrieved. From 142 patients, pre-surgical imaging had a sensitivity of 76.4%, specificity of 73.7%, positive predictive value (PPV) of 94.9%, and negative predictive value (NPV) of 32.6% (p value < 0.0001) for detection of muscle invasion. Also, for detection of positive lymph nodes, imaging had a sensitivity of 31.8%, specificity of 85.7%, PPV of 50%, and NPV of 73.7% (p value: 0.022). 44.4% of study population were upstaged after surgery (24.6% associated with N-upstaging) and 18.3% were downstaged (12% associated with N-downstaging). Receipt of neoadjuvant chemotherapy and T-stage were not correlated with N-upstaging. On multivariate analysis, lymphovascular invasion (LVI) maintained its significance for independent prediction of upstaging (OR 3.3, 95% CI 1.5-7.5, p value: 0.004) and inversely with downstaging (OR 0.34, 95% CI 0.12-0.96, p value: 0.04). Older age (OR 1.03, 95% CI 1.0-1.05, p value 0.047), positive margins (OR 2.1, 95% CI 1.2-3.8, p value 0.011), presence of LVI (OR 2.5, 95% CI 1.4-4.7, p value 0.003), perineural invasion (OR 2.0, 95% CI 1.2-3.4, p value 0.013), and lymph node ratio (OR 1.011, 95% CI 1.001-1.021, p value 0.03) were associated with worse survival. Also, N-upstaging independently predicted a worse survival after controlling for surgical pathology stage (OR 2.3, 95% CI 1.2-4.5, p value 0.011). The optimal target volume in definitive chemoradiotherapy of node-negative bladder cancer patients remains to be established. Since then, customizing the treatment is considered especially for positive LVI in TURBT specimen.