Expression Analysis of Same-Patient Metachronous and Synchronous Upper Tract and Bladder Urothelial Carcinoma.

PURPOSE: We compared upper tract urothelial carcinoma (UTUC) and bladder urothelial carcinoma (BUC) in same-patient metachronous UTUC and synchronous UTUC and BUC using next-generation sequencing. MATERIALS AND METHODS: Consecutive untreated same-patient samples of UTUC and BUC were macrodissected from unstained formalin-fixed, paraffin-embedded slides after quality control. Samples were divided into 4 groups: 1) UTUC-metachronous BUC, 2) BUC-metachronous UTUC, 3) synchronous UTUC-BUC, 4) UTUC without BUC. Exclusions were inadequate clinical data or histological tumor purity <30%. Whole transcriptome RNA sequencing was performed. After quality assessment, gene expression clusters using unsupervised hierarchical consensus clustering and correlation with pertinent clinicopathologic variables, a prior RNASeq data set and other published data were performed. RESULTS: RNAseq was performed on 95 samples (UTUC=61, BUC=34) from 40 untreated patients. Unsupervised consensus clustering segregated the tumors into 2 clusters that were enriched with BASE47 basal-like or luminal-like gene expression. Almost two-thirds (61.9%) of Group 2 tumors were basal-like, while the majority of Groups 1, 3, 4 (80.6%, 70.0% and 69.6%, respectively) were luminal-like (p=0.017). Further analyses revealed that the differences in basal-like and luminal-like gene expression were associated with differential fibroblast and immune cell gene expression signatures. In all, 87.5% of metachronous tumors maintained subtype membership. CONCLUSIONS: Gene expression analysis of same-patient metachronous UTUC-BUC suggests that the majority of mUTUC developing after BUC appear more basal-like, while synchronous and initial UTUC tumors appear luminal-like. Metachronous tumors largely maintain molecular subtype membership of the initial tumor regardless of chronologic development or anatomical origin.

Case Report: Glucocorticoid Effect Observation in a Ureteral Urothelial Cancer Patient With ICI-Associated Myocarditis and Multiple Organ Injuries.

Immune checkpoint inhibitor (ICI)-associated immune-related adverse events (irAEs) are becoming important safety issues worthy of attention despite the exciting therapeutic prospects. The growing development of new ICIs also brings new cases of irAEs, raising more challenges to clinicians. Cardiac injury is rare but life-threatening among diverse organ injuries, and effective interventions are critical for patients. Here, we report a novel programmed cell death protein-1 (PD-1) inhibitor tislelizumab-associated severe myocarditis and myositis accompanied by liver and kidney damage in a ureteral urothelial cancer patient, who was firstly treated by cardiologists because of cardiac symptoms. Due to the lack of experience about ICI-associated irAEs, an initial low-dose (0.5 mg/kg/day) and short-term methylprednisolone therapy was used and found to be ineffective and risky to the patient; then, steroid therapy was modulated to a higher dose (1.5 mg/kg/day) with prolonged time course, and improvement of patient symptoms and laboratory markers were observed quickly and persistently. The patient did not show adverse events under this steroid dosage. This case reports a rare tislelizumab-related myocarditis and multiple organ injuries, which provides valuable experience to cardiologists like us. Early recognition of ICI-associated myocarditis and sufficient dosage and time course of glucocorticoid therapy are critical for severe cases. High-quality clinical evidence about the precise diagnosis and therapy in ICI-associated myocarditis and other organ injuries are necessary to guide our clinical works.

Immunosuppression and Malignant Neoplasms: Risk-Benefit Assessment in Patients with Inflammatory Bowel Disease.

BACKGROUND The treatment of inflammatory bowel disease aims to induce and maintain disease remission, avoid complications, and restore quality of life. The treatments include the use of immunosuppressants and biological therapy. Despite the effectiveness of these treatments in controlling disease activity and in limiting complications, there remains an increased risk of developing malignancies. CASE REPORT A 70-year-old male patient with ulcerative colitis who had pancolitis was initially treated with mesalazine. In 2010, the medication was changed to azathioprine due to clinical disease activity. The patient demonstrated clinical and endoscopic response to the medication, but presented recurrent facial lesions identified as non-melanoma skin cancer in 2014, 2015, and 2016. Azathioprine was discontinued and anti-TNF therapy was started, but no satisfactory clinical or endoscopic response was observed. The patient developed hematuria and a ureter tumor was found with subsequent ureteronephrectomy. Moreover, the patient underwent total colectomy with ileostomy as a treatment for refractory ulcerative colitis. CONCLUSIONS Immunosuppressive therapy can facilitate the development of malignant neoplasms, accelerate tumor growth, and favor the onset of metastases. The types of tumors most associated with its use are lymphoproliferative tumors and non-melanoma skin cancer. The benefits of adequate control of inflammatory bowel disease are clear and the use of immunosuppressants should not be limited by these potential adverse outcomes; however, the risk-benefit profile of immunosuppression should always be assessed on a case-by-case basis.

Association of cancer progression with elevated expression of programmed cell death protein 1 ligand 1 by upper tract urothelial carcinoma and increased tumor-infiltrating lymphocyte density.

BACKGROUND: Increased expression of programmed cell death 1 ligand 1 (PD-L1) by tumor cells is thought to be a mechanism through which solid cancers promote immune tolerance. However, the association between PD-L1 expression and the prognosis of upper urinary tract urothelial carcinoma (UTUC) remains unknown. METHODS: We examined immunohistochemical PD-L1 expression and the tumor-infiltrating lymphocyte density (TILD) in 79 patients with UTUC who underwent nephroureterectomy. We classified the tumors into four types based on the combination of PD-L1 expression and TILD, and studied the clinicopathological characteristics of these four tumor types. RESULTS: Elevated expression of PD-L1 by tumor cells and a higher TILD were associated with a worse histological grade, higher pT stage, and higher peripheral blood neutrophil-to-lymphocyte ratio. Elevated expression of PD-L1 by tumor cells, a higher TILD, and type I, III, or IV tumors with elevated expression of either PD-L1 or TILD showed a positive correlation with poorer differentiation and local invasion. These three variables were associated with shorter progression-free survival and overall survival in univariate analysis, but only the latter was an independent determinant according to multivariate analysis. The patients who had type II tumors with lower PD-L1 expression and a lower TILD showed more favorable survival than the other three groups. CONCLUSIONS: These findings suggest that PD-L1 expression and TILs in the tumor microenvironment influence the progression of UTUC. Accordingly, it is important to understand the immunologic characteristics of the tumor microenvironment to develop more effective treatment strategies for this cancer.

Upper tract urothelial carcinoma has a luminal-papillary T-cell depleted contexture and activated FGFR3 signaling.

Upper tract urothelial carcinoma (UTUC) is characterized by a distinctly aggressive clinical phenotype. To define the biological features driving this phenotype, we performed an integrated analysis of whole-exome and RNA sequencing of UTUC. Here we report several key insights from our molecular dissection of this disease: 1) Most UTUCs are luminal-papillary; 2) UTUC has a T-cell depleted immune contexture; 3) High FGFR3 expression is enriched in UTUC and correlates with its T-cell depleted immune microenvironment; 4) Sporadic UTUC is characterized by a lower total mutational burden than urothelial carcinoma of the bladder. Our findings lay the foundation for a deeper understanding of UTUC biology and provide a rationale for the development of UTUC-specific treatment strategies.

Predictive value of preoperative lymphocyte-to-monocyte ratio for patients with upper tract urothelial carcinoma.

BACKGROUND: To determine the predictive value of preoperative lymphocyte-to-monocyte ratio (LMR) for pathological and survival outcomes in upper tract urothelial carcinoma (UTUC) after radical nephroureterectomy (RNU). METHODS: This retrospective study included 704 UTUC patients between 2008 and 2017. We used a cutoff LMR of 3.6 to evaluate its relationship with oncological outcomes after RNU, using the Kaplan-Meier method and logistic regression models. RESULTS: During median follow-up of 39 months, decreased preoperative LMR was an independent predictor of poor pathological outcomes. Kaplan-Meier analysis revealed that patients with low LMR (<3.6) had poor cancer-specific survival (CSS), recurrence-free survival (RFS), and overall survival (OS); this prognostic value was ascertained for patients with high pathological grade and more advanced stage UTUC, but not for patients with low pathological features. Multivariate Cox proportional hazards model revealed that low LMR was an independent predictor of poor CSS, RFS and OS [hazard ratio (HR) = 1.42, P = 0.02; HR = 1.39, P = 0.008; HR = 1.38, P = 0.017, respectively]. CONCLUSION: Preoperative low LMR was an independent predictor of poor pathological and oncological outcomes of UTUC after RNU. Subgroup analysis confirmed that the prognostic value of LMR was confined to patients with a high pathological grade and more advanced stage tumor.

The role of macrophages in the differentiation process of ureteral polyps.

Objective To evaluate the role of macrophage infiltration in the differentiation process of ureteral polyps and cancers. Methods This retrospective immunohistochemical study analysed archival samples of pathologically-confirmed specimens of low- and high-grade ureteral cancer, ureteral papilloma and ureteral polyps. The samples were immunohistochemically stained for cluster of differentiation (CD)4, CD8, CD16, CD25, CD56 and CD68 using immunofluorescence in order to identify different T-lymphocyte populations and macrophages. Results A total of 70 specimens were included in the analysis: 21 specimens of ureteral cancer, 17 specimens of ureteral papilloma, and 32 specimens of ureteral polyps. The largest proportion of CD4+CD25+ regulatory T cells was observed in the low-grade ureteral cancer group and almost none were observed in ureteral papillomas. The largest proportion of CD8+ cytotoxic T-lymphocytes was observed in the ureteral polyps. The largest proportion of CD56+ natural killer cells was detected in the ureteral polyps, with very low levels observed in the other three groups. The largest proportion of CD16+CD68+ macrophages was observed in the high-grade ureteral cancer group, which was significantly higher than that observed in the ureteral papillomas. Conclusions This study revealed that CD16+CD68+ macrophages appear to participate in ureteral neoplastic transformation.

Unique expression pattern of viral proteins in human herpesvirus 8-positive plasmablastic lymphoma: a case report.

BACKGROUND: Human herpesvirus 8 (HHV8)-positive plasmablastic lymphoma is a disease which correlates with acquired immunodeficiency syndrome (AIDS). Little is known about the pathogenesis of the disease due to its rarity. We report an autopsy case about AIDS related HHV-8-positive plasmablastic lymphoma and presents an examination about HHV8 related proteins for the disease by using immunohistochemical techniques. CASE PRESENTATION: Two kinds of tumors complicated the male AIDS patient: one was HHV-8-positive plasmablastic lymphoma and the other was Kaposi's sarcoma (KS). Immunohistochemically, the lymphoma cells were positive for HHV8-associated lytic early proteins as well as HHV8 latency-associated nuclear antigen 1 (LANA-1), and, on the other hand, the lymphoma cells were negative for lytic immediately early proteins. KS was positive for only LANA-1. CONCLUSION: These findings indicate that the lymphoma cells acquired an ability to proliferate without de novo HHV8 replication. Moreover, the onset mechanisms of HHV-8-positive plasmablastic lymphoma may be different from those of KS.

Validation of pretreatment neutrophil-lymphocyte ratio as a prognostic factor in a European cohort of patients with upper tract urothelial carcinoma.

OBJECTIVE: To investigate the potential prognostic significance of the neutrophil-lymphocyte ratio (NLR) in a large European cohort of patients with upper urinary tract urothelial cell carcinoma (UUT-UCC). PATIENTS AND METHODS: We retrospectively evaluated data from 202 consecutive patients with non-metastatic upper urinary tract urothelial cell carcinoma (UUT-UCC), who underwent surgery between 1990 and 2012 at a single tertiary academic centre. Patients' cancer-specific survival (CSS) and overall survival (OS) were assessed using the Kaplan-Meier method. To evaluate the independent prognostic significance of the NLR, multivariate proportional Cox regression models were applied for both endpoints. RESULTS: A higher NLR was significantly associated with shorter CSS (P = 0.002, log-rank test), as well as with shorter OS (P < 0.001, log-rank test). Multivariate analysis identified a high NLR as an independent prognostic factor for patients' CSS (hazard ratio 2.72, 95% CI 1.25-5.93, P = 0.012), and OS (hazard ratio 2.48, 95% CI 1.31-4.70, P = 0.005). CONCLUSIONS: In the present cohort, patients with a high preoperative NLR had higher cancer-specific and overall mortality after radical surgery for UUT-UCC, compared with those with a low preoperative NLR. This easily identifiable laboratory measure should be considered as an additional prognostic factor in UUT-UCC in future.

Phase I clinical study of anti-apoptosis protein survivin-derived peptide vaccination for patients with advanced or recurrent urothelial cancer.

Survivin, a member of the inhibitor of apoptosis protein family, is expressed in many malignant tumors including urothelial cancer but is hardly detectable in normal, differentiated adult tissues. Previously we reported CD8-positive cytotoxic T-lymphocytes (CTLs) were successfully induced by stimulation with survivin-2B80-88 peptide in vitro. We started a phase I clinical study of survivin-2B80-88 peptide vaccination for advanced urothelial cancer patients to assess the safety and efficacy of this vaccination. Nine patients were received vaccination and were evaluated for immunological evaluation, adverse events, and clinical responses. A total of 46 vaccinations were carried out. There was no severe adverse event. HLA-A24/survivin-2B80-88 peptide tetramer analysis revealed a significant increase in the peptide-specific CTL frequency after the vaccination in five patients. Slight reduction of the tumor volume was observed in one patient. Survivin-2B80-88 peptide-based vaccination is safe and should be further considered for potential immune and clinical efficacy in urothelial cancer patients.

Identification of immunohistochemical factors that predict the synchronous or metachronous development of bladder tumors in patients with upper urinary tract tumors.

OBJECTIVE: To identify markers that predict the synchronous or metachronous development of bladder cancer in patients with upper urinary tract (UUT) tumors. MATERIALS AND METHODS: Between March 2001 and December 2005, we identified 38 consecutive patients who had been histologically diagnosed as having transitional cell carcinoma in the renal pelvis and ureter. These patients were divided into 2 groups (n = 19 per group): group 1 patients with metachronous or synchronous bladder cancer, and group 2 patients with UUT tumors only. We analyzed the differences between the 2 groups with respect to the expression of various biomarkers (p53, Rb, Ki-67, PTEN, and bcl-2) and in terms of clinical parameters. RESULTS: The 2 groups differed significantly in terms of multiplicity (p = 0.029), papillary configuration (p = 0.001), the presence of lymphovascular emboli (p = 0.019), and Ki-67 overexpression (p = 0.029) in UUT tumors. Multivariate analysis revealed that Ki-67 overexpression in UUT tumor tissues significantly predicts bladder cancer development (HR 6.440; 95% CI 1.121-37.014; log rank p = 0.037). CONCLUSION: Ki-67 overexpression in UUT tumor tissues was found to be an independent predictor of the development of bladder cancer in UUT tumor patients.

Establishment and characterization of human urothelial cancer xenografts in severe combined immunodeficient mice.

AIM: To establish and characterize a murine xenograft model of human urothelial cancer in severe combined immunodeficient (SCID) mice for therapeutic simulation. METHODS: Pieces of 30 freshly resected urothelial tumors (24 obtained from bladder and 6 from ureter or pelvis) were implanted subcutaneously into SCID mice, and xenograft tumors were passed in tumorigenic cases. At each passage, histopathology, TP53 mutational status assessed by yeast p53 functional assay, and the Ki-67 labeling index (LI) were examined to evaluate the preservation of original features. A growth delay assay after single-dose irradiation was performed in four representative xenografts. RESULTS: Tumor growth was observed in 18 mice (60%, 18/30). Histologically, 15 of the 18 were epithelial carcinomas similar to the original tumors, whereas the other 3 were Epstein-Barr virus-associated lymphoproliferative disease, resulting in a 50% (15/30) take rate. No correlation was found between the tumor take rate and the clinicopathologic features, TP53 mutational status, or Ki-67 LI of the patients' tumors. Of these 15 xenografts, 11 xenografts were passed from 3 to 10 generations. TP53 mutational status remained stable during the passages, and the Ki-67 LI of eight xenografts was within a range of 50% of the LI of the original tumors, although the other three xenografts increased by over 50%. Specific growth delay after irradiation, independent of the original tumor growth speed and Ki-67 LI, was observed in four xenografts. CONCLUSIONS: SCID mice are useful recipients for investigations of human urothelial cancer with a wide biological range. This easy-to-handle xenograft system can help to develop a better in vivo preclinical evaluation system for therapeutic agents as well as the investigation of tumor pathophysiology.

[The prognostic value of KI-67 proliferation index in transitional cell carcinoma of renal pelvis and ureter].

The investigation deals with Ki-67 immunoreactivity assay in upper urinary tract transitional cell carcinoma (TCC) with respect to grade, stage and survival after radical surgery. In a retrospective study (5yrs) of 37 patients with TCC of the renal pelvis and ureter, who had undergone radical nephroureterectomy and bladder resection, pT1-pT4 lesions and G1-G3 tumors were identified. Ki-67 expression was evaluated by immunohistological staining (1:100; MIB-1; Immunotech. Inc., Westbrook, USA). By using fifteen x600 visual fields, Ki-67 labeling index (number of positive cells per 100 tumor cells) was found (mean +SD--29.7 +/- 9.22). There was a correlation between the index and tumor stage (p < 0.001) and grade (p = 0.002). The Ki-67 values in excess of 27 corresponded to high risk of bladder recurrence (p < 0.001) and short duration of recurrence development (p = 0.067) whereas, for the index of under 22, five-year progression-free survival was more frequent (p < 0.001). Having been tested in that study, discriminative modeling yielded the following parameters: sensitivity and specificity for bladder recurrence was 93% and 79% while for 5-year progression-free survival- 89% and 100%.

Papillary adenocarcinoma of the renal pelvis and ureter producing carcinoembryonic antigen, carbohydrate antigen 19-9 and carbohydrate antigen 125.

We report a case of advanced renal pelvis and ureter adenocarcinoma producing carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9) and carbohydrate antigen 125 (CA125). A 72-year-old woman was diagnosed with right renal pelvic and ureter tumor with para-aortic lymph node swelling. Biopsy of the ureteral mass revealed papillary adenocarcinoma. Serum levels of CEA, CA19-9 and CA125 were extremely elevated. The patient was successfully treated with paclitaxel/carboplatin chemotherapy followed by surgery.

[A case of CA19-9 producing transitional cell carcinoma of the ureter].

We report a case of CA19-9 producing urothelial carcinoma of the right ureter. A 61-year-old male patient who had an extremely high value of serum CA19-9 (1,185 U/ml) with right hydronephrosis was referred to us. Magnetic resonance urography and retrograde ureterography revealed a long irregular filling defect in the right distal ureter. Under the diagnosis of right ureteral tumor, we performed right total nephroureterectomy and pelvic lymphadenectomy. The tumor was histologically diagnosed as grade 1 transitional cell carcinoma and pelvic lymphnodes were positive (pT1N2M0). The tumor cells showed positive immunostaining for CA19-9. The serum CA19-9 level was normalized after the operation and successive adjuvant chemotherapy (M-VAC 2 course). No recurrence was found for 15 months after operation. In this case, the serum CA19-9 level was useful as a tumor marker.

Ny-ESO-1 expression and immunogenicity associated with transitional cell carcinoma: correlation with tumor grade.

NY-ESO-1 mRNA expression in transitional cell carcinoma was investigated by reverse transcription-PCR and immunohistochemistry. NY-ESO-1 mRNA was detected in 20 of 62 (32%) tumor specimens. There was a correlation between NY-ESO-1 expression and tumor grade: 0 of 4 (0%) grade 1 (G1), 6 of 26 (23%) grade 2 (G2), and 14 of 32 (44%) grade 3 (G3) tumors were NY-ESO-1 mRNA positive. Immunohistochemical analysis using NY-ESO-1-specific monoclonal antibody ES121 showed that 2 of 14 NY-ESO-1 mRNA-expressing G3 tumors were positive for NY-ESO-1. No NY-ESO-1 staining was observed in the panel of 30 G1 or G2 tumor specimens, including 6 NY-ESO-1 mRNA-positive cases. Sera from an expanded panel of 124 patients with transitional cell carcinoma were tested for the presence of NY-ESO-1 antibody. Seropositivity was observed in 9 of 72 (12.5%) patients with G3 tumors, whereas none of 52 patients with G1 or G2 tumors produced antibody against NY-ESO-1. In the 9 positive patients with NY-ESO-1 antibody, 4 had muscular invasive tumors, and 5 had carcinoma in situ.

Organ-specific (localized) synthesis of Ig light chain amyloid.

Ig amyloidosis is usually a systemic disease with multisystem involvement. However, in a significant number of cases amyloid deposition is limited to one specific organ. It has not been determined if the Ig light chain (LC) amyloid precursor protein in localized amyloidosis is synthesized by circulating plasma cells with targeting of the amyloid fibril-forming process to one specific organ, or whether the synthesis of Ig LC and fibril formation occurs entirely as a localized process. In the present study local synthesis of an amyloid fibril precursor LC was investigated. Amyloid fibrils were isolated from a ureter that was obstructed by extensive infiltration of the wall with amyloid. Amino acid sequence analysis of the isolated fibril subunit protein proved it to be derived from a lambdaII Ig LC. Plasma cells within the lesion stained positively with labeled anti-lambda Ab and by in situ hybridization using an oligonucleotide probe specific for lambda-LC mRNA. RT-PCR of mRNA extracted from the tumor and direct DNA sequencing gave the nucleotide sequence coding specifically for the lambdaII amyloid subunit protein, thus confirming local synthesis of the LC protein.

Targeted diagnosis of bladder and ureteral carcinoma using radiolabelled BDI-1.

The aim of this study is to investigate the possibility of radioimmunoimaging (RII) by radiolabelled anti-bladder carcinoma monoclonal antibody BDI-1 applied to diagnosis of bladder cancer and ureteral cancer. BDI-1 was labelled with 131I and 99mTc. The immunoreactivity, pharmacokinetics and biodistribution in mice were studied. RII was performed in 46 patients. The results showed that 131I, 99mTc-BDI-1 have satisfactory immunoreactivity and excellent tumor-locating properties. The blood clearance half-life T1/2alpha and T1/2beta were 35 h in the first phase and 151 h in the second phase, respectively. Thirty-nine patients were studied by an intravesical administration method; the sensitivity was 90.5%. Seven patients were studied by an intravenous administration method. The RII results of three cases with primary or recurrent bladder cancer and three cases with ureteral cancer were confirmed histologically. RII was negative in one patient with suspected lung metastasis that was shown on radiography. The investigation revealed that RII can be used as an auxiliary method for the detection of bladder cancer and may be valuable for the diagnosis of ureteral cancer.