Branched evolution and intratumor heterogeneity of urothelial carcinoma of the bladder.

293 Background: Genomic characterization of urothelial carcinoma of the bladder (UCB) has begun to reveal significant intertumor heterogeneity when comparing samples from different subjects. As in other malignancies, intratumor heterogeneity, which may allow for tumor evolution and adaption, poses a significant challenge to personalized-medicine strategies. Methods: To examine UCB tumor evolution and heterogeneity, we performed next-generation targeted sequencing on multiple temporally and spatially separated bladder tumors obtained at time of transurethral resection (TUR) and radical cystectomy (RC). Specimens were analyzed using a next-generation, targeted sequencing assay designed to identify point mutations, indels, and copy number alterations in 300 cancer-associated genes. Results: Phylogenetic reconstruction revealed evidence of branched evolutionary growth. Evaluation of multiple tumors from individual subjects identified both shared and unique potential driver mutations. Evidence of convergent phenotypic evolution was detected through analysis of multiple distinct tumors from several subjects. For example, three separate tumors in one subject shared a common PIK3CA mutation (E453K) and had unique second mutations in PIK3CA (E542V, E545K, and E545Q, respectively). In another subject, distinct inactivating mutations of EP300 were identified in two temporally separated tumor samples. Macrodissection of single tumors into non-invasive and invasive components revealed significant intratumor heterogeneity; one case illustrates how analysis of a muscle-invasive TUR specimen could result in undersampling and thereby miss the tumor clone that persisted at time of RC. Conclusions: We demonstrate branched evolution of UCB through genomic analyses of multiple temporally and spatially distinct bladder tumors from individual subjects. Macrodissection of individual tumor samples identified significant intratumor heterogeneity. These concepts may present major challenges to personalized-medicine approaches that rely on sampling of a single tumor at a specific timepoint in the evolution of a patient’s UCB.

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Targeted sequencing of upper tract urothelial carcinoma.

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.4_suppl.309 ◽

2014 ◽

Vol 32 (4_suppl) ◽

pp. 309-309 ◽

Cited By ~ 2

Author(s):

John P. Sfakianos ◽

Philip H. Kim ◽

Gopa Iyer ◽

Eugene K. Cha ◽

Emily C. Zabor ◽

...

Keyword(s):

Urothelial Carcinoma ◽

Renal Pelvis ◽

Upper Tract Urothelial Carcinoma ◽

Targeted Sequencing ◽

High Grade ◽

Next Generation ◽

Bladder Tumors ◽

P Values ◽

Exact Test ◽

Upper Tract

309 Background: Regardless of the anatomic location (renal pelvis or ureter (upper tract), bladder or urethra) urothelial carcinoma is considered a single entity. Outcomes differ depending on site of initial diagnosis. Moreover, little is known about the comparative biology across primary sites. To define the genomic profile of urothelial carcinoma arising from the upper tract, we used a next generation sequencing technology to analyze 26 sporadic high-grade urothelial tumors of the renal pelvis. Methods: Frozen tumor samples and matched germline blood or normal kidney DNA from 26 nephroureterectomy specimens were obtained under an IRB-approved protocol. DNA was analyzed using a next generation, targeted sequencing assay. We compared these tumors to a set of 108 bladder tumors. Fisher’s exact test analyzed associations with pathological stage and Cox regression with log-rank p-values examined associations with bladder recurrence and disease-free survival. Fisher’s exact test was used to compare mutation frequencies between upper tract and bladder tumors. P-values were adjusted for multiple comparisons using the Benjamini-Hochberg method. Results: Surgical pathology was Ta, T1, T2 and T3 in 7 (26.9%), 5 (19.2%), 4 (15.4%) and 10 (38.5%) patients, respectively. High-grade disease was found in 24 (92.3%) patients with 6 (27.3%) patients having node positive disease. The most frequently altered genes included FGFR3 (42.3%), KDM6A (38.5%), MLL2 (26.9%), TSC1 (26.9%), CREBBP (23.1%), CDKN2A (19.2%) and HRAS (19.2%). No significant differences in tumor and patient characteristics were found between the bladder and upper tract cohorts. A significant difference in the frequency of alterations between the upper tract and bladder cohorts was found in HRAS (19.2% vs 2.8%, p=0.032), TP53 (7.7% vs 57.4%, p<0.001) and TSC1(26.9% vs 5.6%, p=0.024). Conclusions: We identified unique patterns of genomic alterations within upper tract urothelial carcinoma compared to bladder cancers. Upper tract tumors are characterized by a higher frequency of activating FGFR3 and HRAS mutations, TSC1 nonsense mutations, and a lower frequency of TP53 alterations. These findings may have future implications on the utility of targeted therapies in this disease subtype.

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MP21-19 BRANCHED EVOLUTION AND INTRATUMOR HETEROGENEITY OF UROTHELIAL CARCINOMA OF THE BLADDER

The Journal of Urology ◽

10.1016/j.juro.2014.02.847 ◽

2014 ◽

Vol 191 (4S) ◽

Author(s):

Eugene Cha ◽

John Sfakianos ◽

Hikmat Al-Ahmadie ◽

Sasinya Scott ◽

Philip Kim ◽

...

Keyword(s):

Urothelial Carcinoma ◽

Intratumor Heterogeneity ◽

Carcinoma Of The Bladder ◽

Branched Evolution

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Detection of PD-L1 in the urine of patients with urothelial carcinoma of the bladder

Scientific Reports ◽

10.1038/s41598-021-93754-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Georgi Tosev ◽

Wasilijiang Wahafu ◽

Philipp Reimold ◽

Ivan Damgov ◽

Constantin Schwab ◽

...

Keyword(s):

Bladder Cancer ◽

Urothelial Carcinoma ◽

Locally Advanced ◽

Invasive Bladder Cancer ◽

Therapeutic Monitoring ◽

Tumor Evolution ◽

Muscle Invasive Bladder Cancer ◽

Programmed Death ◽

Carcinoma Of The Bladder ◽

Muscle Invasive

AbstractThere are currently five programmed death-1 (PD-1)/programmed death ligand-1 (PD-L1) inhibitors approved for the treatment of locally advanced or metastatic urothelial carcinoma (UC) of the bladder. For platinum-ineligible patients, testing of tumor specimens for PD-L1 expression is required. However, scoring of PD-L1 immunohistochemistry is complex due to different antibodies used, the requirement to score expression in different cellular compartments and intratumoral heterogeneity. It can also be difficult to obtain and test longitudinal tumor samples, which would be desirable to monitor treatment responses and tumor evolution under treatment-induced selective pressure. In the present proof-of concept study, we provide evidence that PD-L1 can be detected in the urine of patients with non-muscle invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC). Urine PD-L1 levels were significantly higher in NMIBC and MIBC patients when compared to patients with various non-malignant urological diseases. Further prospective and independent studies are required to assess the value of PD-L1 in the urine as a novel biomarker with potential for the early detection, prediction and therapeutic monitoring of patients with UC of the bladder.

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Tumor evolution and intratumor heterogeneity of an epithelial ovarian cancer investigated using next-generation sequencing

BMC Cancer ◽

10.1186/s12885-015-1077-4 ◽

2015 ◽

Vol 15 (1) ◽

Cited By ~ 49

Author(s):

Jung-Yun Lee ◽

Jung-Ki Yoon ◽

Boyun Kim ◽

Soochi Kim ◽

Min A Kim ◽

...

Keyword(s):

Ovarian Cancer ◽

Next Generation Sequencing ◽

Epithelial Ovarian Cancer ◽

Intratumor Heterogeneity ◽

Tumor Evolution ◽

Next Generation ◽

Generation Sequencing

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Differentiating Nonfunctional Paraganglioma of the Bladder from Urothelial Carcinoma of the Bladder: Pitfalls and Breakthroughs

BioMed Research International ◽

10.1155/2019/1097149 ◽

2019 ◽

Vol 2019 ◽

pp. 1-7

Author(s):

Musa Male ◽

Tao Ye ◽

Jin Tao ◽

Zhi-qiang Chen ◽

Ejun Peng

Keyword(s):

Urothelial Carcinoma ◽

Preoperative Diagnosis ◽

Clinical Characteristics ◽

Diagnostic Value ◽

Tumor Diameter ◽

Bladder Tumors ◽

Significant Difference ◽

Maximum Tumor Diameter ◽

Carcinoma Of The Bladder ◽

New Onset

Background. Although both nonfunctional paraganglioma of the bladder (NPB) and urothelial carcinoma of the bladder (UCB) are subtypes of bladder tumors, they are entirely different entities with distinct tissue origins and anatomical locations. However, NPB is frequently misdiagnosed as UCB chiefly due to the similarities in their clinical characteristics and cystoscopic features. This study aimed to compare the differences in their clinical characteristics and cystoscopic features. Patients and Methods. Between April 2007 and September 2017, 14 patients with NPB (NPB group) were retrieved from 2 centers, and 42 patients with new-onset UCB (UCB group) were randomly retrieved. Demographic, symptomatic, imaging, and cystoscopic data of patients in both groups were collected and compared. Results. NPB group comprised 7 males and 7 females, with a mean age of 43.1 ± 13.6 years. Compared with the UCB group, patients in the NPB group were significantly younger (p<0.001), less likely to be male (p<0.05), and to present with hematuria (p<0.01). However, no significant difference in maximum tumor diameter was observed between the 2 groups (p=0.609). Compared with the UCB group, cystoscopically, patients in the NPB group were significantly more likely to present with hypervascularization but less likely to present with hemorrhage, necrosis, calcification, pedunculation, and multilesion (p<0.05). No patients with NPB were clinically diagnosed correctly before cystoscopy. Of the 5 patients who underwent both cystoscopy and biopsy, 4 were diagnosed with NPB, while 1 remained undiagnosed. Of the remaining 9 patients who underwent cystoscopy only, 5 were diagnosed with nonepithelial tumor, and 4 were misdiagnosed with UCB. Conclusions. Age, sex, and hematuria may provide clues to differentiating NPB from UCB. Differences in cystoscopic features between NPB and UCB are of high diagnostic value. Cystoscopic biopsy should be considered in the preoperative diagnosis of NPB.

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Patient derived xenografts of upper tract urothelial carcinoma: A potential tool for personalized medicine.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.6_suppl.344 ◽

2017 ◽

Vol 35 (6_suppl) ◽

pp. 344-344

Author(s):

Kwanghee Kim ◽

Katie S Murray ◽

Aditya Bagrodia ◽

Francois Audenet ◽

Sylvia Jebiwott ◽

...

Keyword(s):

Personalized Medicine ◽

Urothelial Carcinoma ◽

Cell Lines ◽

Genetic Fidelity ◽

Genetic Characteristics ◽

Upper Tract ◽

Nsg Mice ◽

Exon Capture ◽

Carcinoma Of The Bladder ◽

Pdx Models

344 Background: Historically, upper tract urothelial carcinomas (UTUC) are treated similarly to urothelial carcinoma of the bladder (UCB). However, compared to UCB, UTUC demonstrates a more aggressive clinical course which may be explained by significant differences in mutational frequencies between UTUC and UCB that were reported using a customized exon capture sequencing assay (the MSK-IMPACT assay). A major limitation in the advancement of the UTUC field is the lack of appropriate models specific to the disease. The objective of this study was to develop and evaluate preclinical models that would recapitulate treatment responses observed in patients. Methods: 35 surgical specimens from nephroureterectomy of patients with UTUC were implanted into immunocompromised NOD-SCID IL2Rg−/− (NSG) mice. The histological and the genomic characterization of patient tumors and PDXs were examined by a board certified pathologist and MSK-IMPACT assay, respectively. Cell lines were also established to assess histologic and genetic fidelity. Chemosensitivity of PDX models was assessed using a 4 week cycle of gemcitabine/cisplatin (or carboplatin) administration and analysis of tumor growth was performed using a Two-way ANOVA test. Results: 12 patient-derived xenograft (PDX) models were established with a success rate of 34% (12/35) and a 14% (3/21) success in developing cell lines. Both models were highly reflective of their original tumors in terms of histology and genomic characteristics in mutation status and copy number alterations. For a representative PDX, chemosensitivity experiments identified gemcitabine/carboplatin as a potentially effective combination, which was also used in the clinical scenario with a therapeutic response. Chemosensitivity of additional PDXs is continuing to be investigated to potentially guide post-operative decision making for treatment of the patient’s clinically. Conclusions: We developed a cohort of stable PDX models and cell lines for UTUC that maintains the genetic characteristics of the patient’s initial tumor. The continued development of these models may facilitate personalized medicine strategies in the treatment of UTUC.

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