Subtypes Of Urothelial Carcinoma Research Articles

CDCP1 expression is frequently increased in aggressive urothelial carcinoma and promotes urothelial tumor progression

The prognosis of patients with advanced urothelial carcinoma (UC) remains poor and improving treatment continues to be a major medical need. CUB domain containing protein 1 (CDCP1) is a known oncogene in various types of solid cancers and its overexpression is associated with impaired prognosis. However, its role in UC remains undetermined. Here we assessed the clinical relevance of CDCP1 in two cohorts of UC at different stages of the disease. Immunohistochemistry showed that CDCP1 is highly expressed in advanced UC, which significantly correlates with shorter overall survival. Importantly, the basal/squamous UC subtype showed significantly enriched CDCP1 at the mRNA and protein levels. The functional role of CDCP1 overexpression was assessed taking advantage of ex vivo organoids derived from the CDCP1pcLSL/+ transgenic mouse model. Furthermore, CDCP1 knockout UC cell lines were generated using CRISPR/Cas9 technology. Interestingly, CDCP1 overexpression significantly induced the activation of MAPK/ERK pathways in ex vivo organoids and increased their proliferation. Similarly, CDCP1 knockout in UC cell lines reduced their proliferation and migration, concomitant with MAPK/ERK pathway activity reduction. Our results highlight the relevance of CDCP1 in advanced UC and demonstrate its oncogenic role, suggesting that targeting CDCP1 could be a rational therapeutic strategy for the treatment of advanced UC.

Urinary Bladder Masses, Rare Subtypes, and Masslike Lesions: Radiologic-Pathologic Correlation

Urinary bladder masses are commonly encountered in clinical practice, with 95% arising from the epithelial layer and rarer tumors arising from the lamina propria, muscularis propria, serosa, and adventitia. The extent of neoplastic invasion into these bladder layers is assessed with multimodality imaging, and the MRI-based Vesical Imaging Reporting and Data System is increasingly used to aid tumor staging. Given the multiple layers and cell lineages, a diverse array of pathologic entities can arise from the urinary bladder, and distinguishing among benign, malignant, and nonneoplastic entities is not reliably feasible in most cases. Pathologic assessment remains the standard of care for classification of bladder masses. Although urothelial carcinoma accounts for most urinary bladder malignancies in the United States, several histopathologic entities exist, including squamous cell carcinoma, adenocarcinoma, melanoma, and neuroendocrine tumors. Furthermore, there are variant histopathologic subtypes of urothelial carcinoma (eg, the plasmacytoid variant), which are often aggressive. Atypical benign bladder masses are diverse and can have inflammatory or iatrogenic causes and mimic malignancy. © RSNA, 2022 Online supplemental material is available for this article.

Nested Subtype of Urothelial Carcinoma in the Upper Urinary Tract: A Series of 15 Cases.

Nested subtype of urothelial carcinoma (NVUC) of the upper urinary tract is rare. While this subtype has an aggressive behavior in the urinary bladder, little is known about this subtype in the renal pelvis and ureter. In addition, due to NVUC's bland morphology and resemblance to von Brunn nests proliferation, which can be florid in the renal pelvis and ureter, establishing a diagnosis of NVUC in these sites can be difficult, especially on small biopsies. Nineteen cases of NVUC of the renal pelvis and the ureter were identified. One was in-house and 18 cases were sent to the genitourinary pathology consult service. Four cases were excluded due to difficulty in obtaining the slides from other institutions. In all, 8/15 (53.33%) cases showed large nested morphology, 4/15 (26.66%) cases showed mixed large and small nested morphology, and only 3/15 (20%) cases showed predominantly small nested morphology. In all cases, the invasive component was pure nested subtype, and tumors were all unifocal. In 8/15 cases, there was the presence of an overlying low-grade papillary urothelial carcinoma. An associated desmoplastic stromal reaction was none/minimal in 12/15 (80%) cases, with a prominent reaction in the remaining 3/15 (20%) cases. Pathologic stages for the resections were pT1 (n=1), pT2 (n=3), pT3 (n=8), and pT4 (n=1). There was only 1 case with a lymph node metastasis at the time of resection who had pT4 disease. Follow-up was available in 10/15 cases, 7/10 cases showed no recurrences or metastases, and the remaining 3/10 cases later developed urothelial carcinoma of the bladder. In 5/15 cases, follow-up could not be obtained. Similar to NVUC in the urinary bladder, NVUC in the upper tract typically lacks an associated desmoplastic reaction and has by definition bland cytology. In contrast to NVUC of the urinary bladder, NVUC in the upper urothelial tract is: (1) more frequently associated with an overlying papillary urothelial carcinoma; (2) more commonly has a large nested pattern; and (3) appears to behave less aggressively.

1736MO Pure or mixed basal/squamous tumours present decreased outcomes after neoadjuvant chemotherapy in the GETUG-AFU V05 VESPER trial

Molecular subtypes of urothelial carcinoma (UC) may impact the outcomes after neoadjuvant chemotherapy in muscle invasive bladder cancer (MIBC). Previous studies reported contradictory results in this setting. Here, we assessed pathological response and 3-year (yr) progression free survival (PFS) according to the consensus molecular subtypes for patients in the French prospective Vesper clinical trial (NCT01812369).

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.

Immunohistochemistry subtyping of urothelial carcinoma is feasible in the daily practice

The preferred treatment of choice in muscle-invasive bladder cancer (MIBC) is usually transurethral resection followed by cystectomy, with neoadjuvant chemotherapy being a second option. As the treatment is associated with relevant side effects, a great effort is being made to improve the selection of patients, with molecular subtyping being one of the main strategies. Our aim was to develop an immunohistochemical algorithm for subtyping MIBCs. After a literature review, we have developed a simple algorithm to subtype MIBCs based on their morphology and three common antibodies: GATA3, CK5/6, and p16. We applied it to 113 muscle-invasive carcinomas. The positivity threshold for GATA3 and CK5/6 was 20% with at least moderate intensity, while p16 was 70% with moderate to intense nuclear and cytoplasmic staining. Cases GATA3 + CK5/6 - were considered luminal, while cases GATA3 - CK5/6 + were classified as nonluminal/basal squamous. Luminal p16 + cases were labeled as genomically unstable and luminal p16 - as Uro-like. Cases GATA3 + CK5/6 + with a predominantly basal pattern were labeled luminal, while diffuse cases were labeled nonluminal/basal squamous. All GATA3-CK5/6 - cases were considered nonluminal and were divided into mesenchymal-like or neuroendocrine, depending on the morphology. We were able to classify the 113 cases as: 82 (72.57%) were luminal, being 47 Uro-like (41.59%) and 35 (30.97%) genomically unstable; 31 (27.43%) were nonluminal, being 24 basal/squamous (21.24%), two (1.76%) mesenchymal-like, and five (4.42%) neuroendocrine like. We have achieved a feasible and cost-effective algorithm to subtype MIBCs from morphological features and the use of three common antibodies. Further studies in external cohorts are necessary to validate these results.

A classification system for urothelial carcinoma (UC) defined by genomic drivers and the tumor microenvironment (TME) is predictive of immunotherapy response.

4591 Background: UC is associated with high recurrence rates, progression, and resistance to platinum-based therapy. Checkpoint inhibitors (CPIs) are often used for treating UC, but predictive biomarkers that characterize response are lacking in the majority of patients. Defining the TME is essential to understanding patient response to CPIs. Employing a transcriptome-based classification platform, we sought to identify predictive and prognostic subtypes of UC using malignant cell and TME features. Methods: We collected a metacohort of 2,418 UC samples from 14 publicly available datasets, one of which had atezolizumab response data (IMvigor210). Using the methodology described in Bagaev et al. 2021, we selected 28 signatures composed of specific gene sets reflecting distinct cellular processes. Analysis of signature expression and unsupervised clustering was performed. Results: We identified 7 recurring novel UC subtypes (Table 1) with unique genomic and molecular characteristics. The subtypes and key findings include: an immune desert (D) subtype characterized by genomic instability high HER2 expression; an immune desert, FGFR-altered (D-FGFR) subtype with FGFR alterations; an immune enriched (IE) subtype with an enriched TME and high CPI response; a fibrotic (F) subtype with a mesenchymal TME, and strong TGFβ signaling; an immune enriched, fibrotic (IE-F) subtype with a mixed TME and a high CPI response rate; a fibrotic, basal (F-B) subtype with a mesenchymal TME and minimal genomic targets, and a neuroendocrine-like (NE-L) subtype with a high CPI response rate. Conclusions: UC can be classified into 7 subtypes with distinct prognoses, CPI response rates, and druggable targets using malignant cell and TME profiling. Patients with IE, IE-F, and NE-L UCs may be good candidates for CPIs. D UCs may benefit from HER2-i and PARP-i, while FGFR-i might be more suitable for D-FGFR UCs. TGFβi and PARPi may be effective for F UCs, but F-B UCs have no targetable findings. These findings warrant additional investigation for clinical translation. [Table: see text]

Tumor-immune microenvironment revealed by Imaging Mass Cytometry in a metastatic sarcomatoid urothelial carcinoma with a prolonged response to pembrolizumab.

Sarcomatoid urothelial carcinoma (SUC) is a rare subtype of urothelial carcinoma (UC) that typically presents at an advanced stage compared to more common variants of UC. Locally advanced and metastatic UC have a poor long-term survival following progression on first-line platinum-based chemotherapy. Antibodies directed against the programmed cell death 1 protein (PD-1) or its ligand (PD-L1) are now approved to be used in these scenarios. The need for reliable biomarkers for treatment stratification is still under research. Here, we present a novel case report of the first Imaging Mass Cytometry (IMC) analysis done in SUC to investigate the immune cell repertoire and PD-L1 expression in a patient who presented with metastatic SUC and experienced a prolonged response to the anti-PD1 immune checkpoint inhibitor pembrolizumab after progression on first-line chemotherapy. This case report provides an important platform for translating these findings to a larger cohort of UC and UC variants.

Molecular pathology of the non-luminal Ba/Sq-like and Sc/NE-like classes of urothelial tumours: An integrated immunohistochemical analysis

Several groups have during past years produced molecular classification schemes for bladder cancer. Even though no consensus on how to define a subtype exists, one approach has been to base definitions on how tumours cluster according to their mRNA expression profiles. In many cases, obtained profiles, and thus class defining features, are affected by signals from non-tumour cells within the biopsy. To overcome this issue, we combined gene expression analyses with analyses of the actual tumour cells by extensive immunohistochemistry (IHC). By this approach we were able to define tumour cell phenotypes i.e., subtypes defined by features of the tumour cells only, and adjust mRNA-based algorithms accordingly. In the present investigation we address the non-luminal Basal/Squamous-like (Ba/Sq) and Small cell/Neuroendocrine-like (Sc/NE) categories of tumours defined by mRNA-based classification. We make use of IHC data for 15 proteins, all known to be instrumental for defining molecular subtypes of urothelial carcinoma. We show that the UroB type of tumours, frequently grouped together with Ba/Sq, are different from the Ba/Sq entity at several essential features and is a derivative of Urothelial-like tumours (Uro). We show that the Sc/NE tumours are similar to but represents extreme versions of Genomically Unstable (GU) tumours. We apply clustering to 423 cases representing all subtypes using IHC data for 14 proteins and show that the obtained grouping conforms well with the mRNA-based classification. This work describes in detail the molecular pathology of non-luminal RNA-based bladder cancer subtypes and highlight similarities/dissimilarities suggestive of origin.

Current Advances in Immune Checkpoint Inhibition and Clinical Genomics in Upper Tract Urothelial Carcinoma: State of the Art.

Upper tract urothelial carcinoma (UTUC) is a rare and challenging-to-treat malignancy. In most patients it is a sporadic tumor entity, less commonly it falls on the spectrum of Lynch syndrome, an autosomal dominant familial tumor syndrome. Localized UTUC with high-risk features as well as the metastatic disease scenario might require systemic therapy. Platinum-based combination chemotherapy is currently the recommended management option. However, the introduction of immune checkpoint inhibitors into the therapeutic armamentarium has led to a paradigm shift in treatment standards. Immunotherapy has been shown to be safe and effective in treating at least metastatic UTUC, although UTUC-specific high-level evidence is still lacking. Recent technological advances and noteworthy research efforts have greatly improved the general understanding of the biological landscape of UTUC. According to the main findings, UTUC represent a particular subtype of urothelial carcinoma frequently associated with activated FGFR3 signaling, a luminal–papillary phenotype and a T-cell-depleted microenvironment. This improved knowledge promises precision oncology approaches that match treatment decision strategies and genomic profile to ultimately result in better clinical outcomes. The aim of this review was to summarize the main currently available evidence on immune checkpoint inhibition and clinical genomics in UTUC.

Molecular determinants of recurrences of the human urothelial tumor

Background. Urothelial carcinoma (UTC) is an aggressive disease with a known propensity for frequent recurrence. It is difficult to predict the velocity of the development of UTC recur using modern means of clinical diagnostics. Therefore, the development of the capabilities of histo-morphological study of tumor tissues is of particular relevance.Materials and methods. The materials of publications (PubMed, CrossRef) for 1990-2021, devoted to the choice of biomarkers for the diagnosis of UTC, the analysis of molecular pathways, progression and metastasis, were studied. The search was carried out for the key phrases "urothelial carcinoma", "recurrent UTK", "stem cells", "biomarkers of bladder cancer", "genetic changes in urothelium", "circulating tumor DNA".Results. Cancer stem cells serve as a source of UTC recurrence after removal from the primary focus, localizing in any areas of the urothelium, as well as outside the main tumor focus and are characterized by a common genotype, but different phenotypic manifestations. To predict the recurrence of the tumour is advisable to use gene expression signatures, since the subtypes of UTC are characterized by clear gene expression profiles. A larger sample and independent dataset is needed to confirm the clinical significance of the findings. Combined biomarkers predict UTC behavior, and FGFR3 and TP53 mutations can be components for a panel for predicting UTC recurrence. The use of the liquid biopsy method with the determination of the level of circulating tumor DNA is a promising diagnostic method that needs to evaluate the results of an initiated randomized trial.Conclusion. The accumulation of knowledge base about the molecular patterns of UTC will help bridge the gap between the results of molecular genetic and clinical diagnostics. Molecular changes in the transitional cell UTC demonstrates a high potential for determining the timing of tumor recurrence, assessing disease-free survival of patients and for planning the resource base of the healthcare system.

The Genitourinary Pathology Society Update on Classification of Variant Histologies, T1 Substaging, Molecular Taxonomy, and Immunotherapy and PD-L1 Testing Implications of Urothelial Cancers.

The Genitourinary Pathology Society (GUPS) undertook a critical review of the recent advances in bladder cancer focusing on important topics of high interest for the practicing surgical pathologist and urologist. This review represents the second of 2 manuscripts ensuing from this effort. Herein, we address the effective reporting of bladder cancer, focusing particularly on newly published data since the last 2016 World Health Organization (WHO) classification. In addition, this review focuses on the importance of reporting bladder cancer with divergent differentiation and variant (subtypes of urothelial carcinoma) histologies and the potential impact on patient care. We provide new recommendations for reporting pT1 staging in diagnostic pathology. Furthermore, we explore molecular evolution and classification, emphasizing aspects that impact the understanding of important concepts relevant to reporting and management of patients.

Cytokeratin 5/6 Phenotypic Expression in Urothelial Bladder Carcinoma: Relation to Clinicopathological Parameters, FAP And CD147 Immunohistochemical Expression

Background: Urothelial bladder carcinoma is a heterogenous disease with variable prognosis. The basal cytokeratin (CK5/6) is associated with basal molecular subtype of urothelial carcinoma and is speculated to be associated with aggressive tumor behavior. Aim: This study aimed to to evaluate immunohistochemical expression of CK5/6, fibroblast activation protein (FAP) and CD147 in urothelial bladder carcinoma in relation to clinicopathological parameters, and to determine clinicopathological characteristics of CK5/6 positive urothelial carcinoma. Material and Methods: the current work included 60 cases of urothelial bladder carcinoma, which were subjected to immunohistochemical staining using CK5/6, FAP and CD147. The association between these markers’ expression and clinicopathological parameters was assessed. Results: Positive expression for CK5/6 was detected in 48.3% of cases. CK5/6 immunoreactivity was significantly related to tumor grade (p-value = 0.002*), muscle invasion (p-value = 0.021*), squamous differentiation (p-value = 0.020*), FAP expression (p-value < 0.001*), and CD147 expression (p-value = 0.001*). No significant association was detected as regard patients' age, sex, or lymphovascular invasion. In turn, FAP expression in tumor microenvironment was significantly related to tumor grade and lymphovascular invasion, but not to muscle invasion or squamous differentiation. In addition, the expression of CD147 showed significant association with tumor grade and muscle invasion. Conclusions: Cytokeratin 5/6 expression in urothelial bladder carcinoma is associated with features of aggressive tumor behavior (in terms of high tumor grade, advanced stage, and squamous differentiation), FAP and CD147 immunoreactivity. Such markers can be considered as progression markers for urothelial carcinoma.

Plasmacytoid Variant of Urothelial Carcinoma: Poor Prognostic Variant with High Expression of CDH1 Mutation

Plasmacytoid variant of urothelial carcinoma is a rare subtype of urothelial carcinoma that has poor prognosis. We describe two cases of patients with the plasmacytoid variant of urothelial carcinoma (PVUC) who had initial response to neoadjuvant chemotherapy followed by radical cystoprostatectomy and lymph node dissection but presented with early relapse and disease progression manifesting with intestinal obstruction and peritoneal carcinomatosis. Tumor genomic sequencing revealed mutations and alterations in ARID1A, CDH1, PIK3CA, RB1 loss, and TERT promoter, as well as tumor mutational burden of 10 Muts/Mb treated with pembrolizumab with a minimal response. A further review of the literature regarding this rare variant is discussed here.

TNM staging towards a personalized approach in metastatic urothelial carcinoma: what will the future be like?-a narrative review.

The American Joint Committee of Cancer (AJCC) tumor-node-metastasis (TNM) classification, with its periodical updates and modifications, has represented and still represents the basis of cancer staging. The historical, long-standing limitations of anatomic-based TNM staging have been recently “threatened” by the impressive amount of data derived from molecular analyses, which have led to an unprecedented level of understanding of cancer genomics. In fact, current era of personalized oncology has witnessed important efforts towards the integration between clinical, anatomical and molecular features; however, despite the promises, personalized oncology faces many obstacles, due to the complex relationship between tumor biomarkers, previously unknown cancer subtypes and clinical and anatomical characteristics. With regard to urothelial carcinoma (UC), the characterization of tumors in large cohorts of patients has provided important information concerning genetic alterations, revealing the presence of biologically relevant subtypes of UC. In the current review, we will provide an overview regarding this recent “translation” from the anatomic-based TNM to a novel horizon, aiming at further “tailoring” personalized oncology, especially focusing on recently published data about the molecular landscape of UC with its therapeutic and prognostic implications.

Role of HER2 and Claudins in Subtypes of Urothelial Carcinoma Identified Bygata3 and Cytokeratin5\6 Immunohistochemical Study

Background: Bladder cancer is globally the ninth, most common malignancy, and the thirteenth most common cause, of cancer death, associated with high morbidity and mortality, if not treated optimally. Bladder cancers can be divided into molecular subtypes, referred to luminal and basal with distinct clinical behaviors. HER2 is one of the established therapeutic targets in many cancers. Claudins are tight junction proteins, known to modulate therapy resistance in cancer cells. Aim: This study aimed to assess Claudins and HER2 status in the context of tumor molecular subtypes, identified by GATA3 and CK5/6 expression, that may help to select urothelialcarcinoma patients, most likely to respond to immunotherapy. Materials and Methods: This retrospective study was done upon 50 cases of conventional urothelialcarcinoma. GATA3, CK5/6, HER2, Claudins1&4 and P53immunostaining were done and correlated with clinico-immuno-pathological parameters. Results: Bladder cancers could be assigned to main intrinsic molecular subtypes, referred to luminal, basal and double negative. Basal & double negative bladder cancers were more aggressive, when compared to luminal cancers. Positive significant statistical correlation was found between HER2, claudin1 and P53 and clinic-immuno-pathologicalparametersas tumor size, grade, TNM stage, LVI, tumor budding and aggressive molecular subtypes (P-value< 0.05). Negative significant statistical correlation was found between claudin4 and fore mentioned clinico-immuno-pathologicalparameters (P-value< 0.05). Conclusions: The molecular subtypes of bladder cancers, HER2, claudin1&4 and P53 can be used for prognostic and therapeutic stratification of bladder cancers patients, and may affect patient outcome.

Machine Learning Based Classification from Whole-Slide Histopathological Images Enables Reliable and Interpretable Diagnosis of Inverted Urothelial Papilloma

Inverted urothelial papilloma (IUP) is a benign neoplasm of the urinary tract that accounts for less than 1% of urothelial tumors. It is diagnostically challenging for pathologists to distinguish histological features of IUP from other subtypes of non-invasive urothelial carcinoma, such as inverted Ta urothelial carcinoma (UCInv) and low-grade Ta urothelial carcinoma (UCLG). Using a machine learning approach, we analyzed the H&E-stained whole-slide histopathological images of 64 IUP (the largest cohort to date), 69 UCInv, and 92 UCLG samples, and propose a reliable, reproducible, and interpretable machine learning pipeline to classify IUP from other non-invasive urothelial carcinomas. The results showed that our method could achieve area under the ROC of 0.913 and 0.920 for classifying IUP from UCInv and UCLG, respectively, which is superior to competing methods, including deep learning-based methods. Testing of the classification models on an external validation dataset confirmed that our model can effectively identify IUP with high accuracy. Our results suggest that the proposed machine learning pipeline can robustly and accurately capture histopathological differences between IUP and other urothelial carcinoma subtypes, which can be extended to identify other rare cancer subtypes with limited samples and has the potential to expand the clinician’s armamentarium for accurate diagnosis. Funding: This work was supported in part by Indiana Precision Health Initiative to Kun Huang and Jie Zhang. Declaration of Interest: None to declare. Ethical Approval: This research was approved by the Institutional Review Boards of participating institutions.

Stratification of urinary bladder carcinoma based on immunohistochemical expression of CK5, CK14 and CK20

ABSTRACT Molecular subtyping of urothelial carcinoma (UC) is similar to that of breast cancer and is based on the developmental biology approach. The aim of the present study is to assess the prognostic impact of CK5, CK14, and CK20 expression in urinary bladder cancer (UBC) with the potential to stratify them into different subtypes. The current study examined the immunohistochemical expression of CK5, CK14, and CK20 in 90 specimens of UBC. CK5 was expressed in 81.1% of the cases and was significantly associated with old age, muscle invasion, presence of bilharziasis, and tendency for poor overall survival. CK20 was expressed in 47.8% of the cases and was associated with nonmuscle invasion and pure UC while 50% of the cases expressed CK14 that were associated with muscle invasion and perineural invasion. Most squamous cell carcinoma and those associated with bilharziasis were belonged to Ck5+/CK20- subgroup while pure UC and those lacked bilharziasis were located in the Ck5+/CK20+ subgroup. The basal group (Ck5+/CK14+/CK20-) showed high proliferative features compared to the intermediate group (Ck5+/CK14-/CK20-). Generally, presence of CK5 is associated with adverse features especially in the group lacking CK20; however, basal and intermediate subgroups share CK5 expression but they show different proliferative capacities, so their distinction by CK14 is helpful.

Review Article: Prognostic Value of CK5/6 Expression in Urinary Bladder Carcinoma

Abstract Background: Bladder cancer in Egypt is the most prevalent malignancy among Egyptian males (16%), producing >7900 deaths annually, which is strikingly higher than most other parts of the world. The genesis and progression of urothelial bladder carcinoma (UBC) is known to involve alterations in several molecular pathways. Subtyping of bladder carcinoma patients based on the molecular alterations in their primary tumors may permit risk stratification and administration of more personalized therapies. Aim of Study: The aim of this study is to review the studies investigating the frequency of CK 5/6 expression and its value in the stratification of urothelial bladder carcinoma in English literature. Material and Methods: The data were retrieved from databases (PubMed, Medscape and Google scholar) and also material available on the Internet from 2001 till 2019. A structured systematic review was performed with the results tabulated. Results: The initial search presented 8 articles (2001- 2019). The frequency of CK 5/6 expression in UBC cases ranged between 19.7% and 62%. Positivity of CK 5/6 showed significant association with female gender, advanced tumor stage and pure SCC and UC with squamous differentiation. Positivity for CK 5/6 showed significant association with shorter overall survival in UC cases with squamous differen-tiation. CK5/6 expression alone was not associated with clinical outcome in any of the articles but showed significant association when combined with other markers. Conclusions: CK 5/6 as a basal marker could identify aggressive subtypes of urothelial carcinoma as those associated with squamous differentiation and advanced stage, however, it is not related to grading.

Abstract 859: Deep learning predicts expression-based molecular subtypes and immune status of urothelial cancer using digital pathology slides

Abstract RNA-sequencing (RNA-seq) has revealed intrinsic subtypes of urothelial carcinoma (UC). We have recently developed a molecular classifier to separate UC tumors to ‘hot' and ‘cold' based on the expression of immune-related genes. Both molecular subtype and immune-status correlate with response to different treatments, however, RNA-seq data is not readily available for all tumors thus limiting our ability to predict treatment response. We hypothesized that a deep learning approach can predict urothelial cancer molecular subtype and classify immune cell levels from pathology tissue slides. To test this hypothesis, we developed a deep learning model to classify urothelial carcinoma subtypes from hematoxylin and eosin (H&amp;E) stained tissue sections using BASE47 luminal-basal classification and our immune-status classifiers based on RNA-seq data from patients in the TCGA dataset as ground truth. Patches of 2048x2048 pixels were generated from 446 whole slide images (WSIs) resulting in a total of 79,053 images. 70% of the patients were used for training, and 15% are being used as a validation set. The remaining 15% of the data will be a blind test set. We used the Resnet-18 architecture, which performed best based on empirical testing (high accuracy while minimizing overfitting). We performed a color normalization step based on the Vahadane algorithm (IEEE Trans Med Imaging. 2016;35(8):1962 71) to analyze the predictive performance and generalizability of the model compared to non-normalized slides. We also used gradient-weighted class activation mapping to highlight the regions of the tissue that may be important for classification. After 20 epochs of training, we achieved a 74% patch prediction accuracy on the validation set with a ROC-AUC of 0.80 for non-normalized immune cell classification. For normalized images, we achieved a 70% patch prediction accuracy on the validation set with a ROC-AUC of 0.75. For the luminal-basal subtype classification task, we achieved a 68% patch wise accuracy on the validation set with a ROC-AUC of 0.76 for normalized data and 68% patch-wise accuracy with a ROC-AUC of 0.74 for non-normalized data. We are currently testing the model on an internal urothelial cancer cohort at Weill Cornell Medicine. Our data demonstrates the utility of deep learning methods in predicting UC molecular subtype and immune status from digital pathology slides. This data has important implications for predicting prognosis and treatment response in UC patients. Citation Format: Matthew Bryan Brendel, Pegah Khosravi, Emily Tse, Lilly Gu, Kyrillus Shohdy, Rohan Bareja, Bhavneet Bhinder, Olivier Elemento, Bishoy Faltas, Iman Hajirasouliha. Deep learning predicts expression-based molecular subtypes and immune status of urothelial cancer using digital pathology slides [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 859.