Dynamic alteration and prognostic significance of tumor-associated CD68+ and CD68+ PD-L1- macrophages in muscle-invasive bladder cancer treated with neoadjuvant chemotherapy.

BACKGROUND: The current study aimed to investigate the dynamic alteration and prognostic significance of tumor-infiltrating lymphocytes (TILs), tumor-associated macrophages (TAMs), and PD-L1 status of immune cells in muscle-invasive bladder cancer (MIBC) treated with neoadjuvant chemotherapy (NAC). METHODS: Multiplex immunofluorescence staining was performed to examine CD68+ TAM, CD4+ T cell, CD8+ T cell, FOXP3+ Treg cell, and PD-L1 expression in paired MIBC tissues (n = 54) before and after NAC. Patients were then divided into definite responders (DR), (≤pT1) and incomplete responders (IR). RESULTS: There was no significant difference between DR and IR cohorts for the immune cell infiltration levels at the baseline status. Tobacco history was identified to be associated with worse NAC efficacy. CD68+ (stroma area: p = 0.025; tumor area: p = 0.028; total area: p = 0.013) and CD68+ PD-L1- (stroma area: p = 0.035; tumor area: p = 0.013 total area: p = 0.014) TAMs infiltration levels decreased significantly after NAC, while there was no significant difference of CD68+ PD-L1+ and TILs. The infiltration of CD68+ (p = 0.033), CD68+ PD-L1- (p = 0.033), and CD68+ PD-L1+ (p < 0.001) TAMs in stroma area were significantly associated with poorer disease-free survival rate (DFS) of MIBC patients. CONCLUSION: CD68+ and CD68+ PD-L1- TAMs infiltration levels decreased significantly after NAC and pre-treatment TAM infiltration levels were independent prognostic factors for MIBC patients. While there was no sufficient evidence demonstrating that pre-treatment TILs or TAMs could predict response to NAC in MIBC patients.

Co-activation of super-enhancer-driven CCAT1 by TP63 and SOX2 promotes squamous cancer progression.

Squamous cell carcinomas (SCCs) are aggressive malignancies. Previous report demonstrated that master transcription factors (TFs) TP63 and SOX2 exhibited overlapping genomic occupancy in SCCs. However, functional consequence of their frequent co-localization at super-enhancers remains incompletely understood. Here, epigenomic profilings of different types of SCCs reveal that TP63 and SOX2 cooperatively and lineage-specifically regulate long non-coding RNA (lncRNA) CCAT1 expression, through activation of its super-enhancers and promoter. Silencing of CCAT1 substantially reduces cellular growth both in vitro and in vivo, phenotyping the effect of inhibiting either TP63 or SOX2. ChIRP analysis shows that CCAT1 forms a complex with TP63 and SOX2, which regulates EGFR expression by binding to the super-enhancers of EGFR, thereby activating both MEK/ERK1/2 and PI3K/AKT signaling pathways. These results together identify a SCC-specific DNA/RNA/protein complex which activates TP63/SOX2-CCAT1-EGFR cascade and promotes SCC tumorigenesis, advancing our understanding of transcription dysregulation in cancer biology mediated by master TFs and super-enhancers.

Distinct subcellular localization of E-cadherin between epithelioid angiomyolipoma and triphasic angiomyolipoma: A preliminary case-control study.

Epithelioid angiomyolipoma (EAML) is a rare variant of angiomyolipoma (AML). Previous studies have demonstrated that epithelial (E-)cadherin is expressed in two subtypes of AML, EAML and triphasic AML; however, the expression pattern of E-cadherin remains unclear. In the present study, a preliminary case-control study was conducted to determine the expression pattern of E-cadherin between EAML and triphasic AML, the control, focusing on the subcellular localization and expression category of E-cadherin. No significant difference was identified in the age, sex, history of tuberous sclerosis, smoking and alcohol consumption between the two groups (P>0.05). In EAML, 9 patients were categorized as exhibiting a low risk of malignant behavior and the other two were categorized as exhibiting an intermediate or high risk of malignant behavior. The proportion of cases expressing E-cadherin, human melanoma black-45 (HMB45), melanoma antigen recognized by T cells 1 (Mart1/Melan A), smooth muscle actin and progesterone receptor were 95.5 (21/22), 95.5 (21/22), 86.4 (19/22), 77.3 (17/22) and 86.4% (19/22), respectively. E-cadherin was identified to be localized, using staining techniques, in the cell membrane and/or cytoplasm. The subcellular localization of E-cadherin was significantly different between EAML and triphasic AML; the majority of EAML cases revealed membranous and cytoplasmic staining, whereas triphasic AML cases demonstrated cytoplasmic staining (P=0.0093). The expression of E-cadherin may be positively associated with HMB45 (P=0.0044) and Mart1/Melan A (P=0.0049). The results of the present study identified that the subcellular localization of E-cadherin may be different between EAML and the control group of triphasic AML. Additionally, E-cadherin and melanocytic markers may be co-expressed in distinct subtypes of AML. A follow-up study with a large sample size to validate the results of the present study, followed by a mechanistic study based on cell lines to determine any significance, are warranted.

A suggestion for pathological grossing and reporting based on prognostic indicators of malignancies from a pooled analysis of renal epithelioid angiomyolipoma.

PURPOSE: Epithelioid angiomyolipoma (EAML) is a rare entity of the kidney. The guideline for grossing and reporting of renal EAML has not been established for Chinese patients. We planned this study to provide some preliminary indicators for draft guidelines of pathological diagnosis among Chinese people. METHODS: The histopathological characteristics of 11 EAML cases from Cancer Hospital, Chinese Academy of Medical Sciences, were reviewed, and a pooled analysis based on our cases and cases from published articles was performed on the histopathological characteristics and prognosis of 56 Chinese patients with EAML. All the cases met the criteria of the 2004 World Health Organization classification of renal tumors. RESULTS: The ratio of female to male was 1.2:1 with the mean age of 43.4 in the 11 cases. All the 11 cases were sampled following the guideline of renal cell carcinoma. The mean tumor size was 6.5 cm. Four (36.4 %) cases showed necrosis. Six (54.5 %) cases showed invasive borders. Only one case showed metastases. In pooled analysis of the total 56 cases with EAML, 10 cases (17.9 %) showed adverse prognosis. Tumor size, necrosis and invasive edge showed significant difference between favorite and adverse prognostic groups (P < 0.05). CONCLUSION: The majority of EAML is benign, and true malignant EAML is rare. The sample of EAML should follow the sample guidelines of renal cell carcinoma with some modifications, emphasizing the presence of necrosis and invading edge. The information of tumor size, necrosis and invasive edge should be included in the diagnostic report of each EAML case.

[Detection of urothelial carcinoma of the urinary bladder by multicolor fluorescence in situ hybridization].

BACKGROUND & OBJECTIVE: Urothelial carcinoma of the urinary bladder is a common malignant neoplasm of the genitourinary system. This study was to analyze chromosome aberrations in urothelial carcinoma of the urinary bladder in Chinese, and evaluate the possibility and validity of multicolor fluorescence in situ hybridization (M-FISH) in detecting urothelial carcinoma of the urinary bladder. METHODS: The probes of chromosome 3, 7, 17 centromeres and 9p21 region were labeled by random primer method. M-FISH was performed on interphase nuclei of 57 samples of urothelial carcinoma to analyze the correlations of chromosome aberration to diagnosis and pathology of urothelial carcinoma. RESULTS: The rate of aneuploidy was 47.4% for chromosome 3, 50.9% for chromosome 7, 56.1% for chromosome 17, and 59.6% for chromosome 9p21 in urothelial carcinoma. All the aberrations had no correlations to tumor stage. The aberrations of chromosomes 3, 7, 17 were significantly correlated to pathologic grade (P<0.01). As using the 4 chromosome probes in combination, the sensitivity for detecting urothelial carcinoma of the urinary bladder was 54.4%. CONCLUSION: M-FISH may be helpful for detecting urothelial carcinoma of the urinary bladder.

Detection of chromosomal alterations in bladder transitional cell carcinomas from Northern China by comparative genomic hybridization.

To identify chromosome alterations in Chinese bladder cancer, forty-six transitional cell carcinomas of the bladder were analyzed by comparative genomic hybridization. Frequent gains of DNA copy number were observed on 1p (13/46), 1q (13/46), 5p (8/46), 6p (9/46), 7p (7/46), 8q (12/46), 11q (8/46), 17q (11/46), 19q (7/46), 20q (8/46) and Yq (8/46), with minimal overlapping regions at 1p32-pter (10/46), 1q21-q24 (12/46), 5p (8/46), 6p22-p23 (7/46), 7p11.2-p14 (7/46), 8q22-q24 (12/46), 11q13-q14 (8/46), 17q22-qter (11/46), 19q11-13.2 (7/46), 20q11-q13.2 (8/46) and Yq11 (8/46). Losses were predominantly found on 2q (16/46), 5q (8/46), 8p (7/46), 9p (8/46), 9q (13/46), 11p (7/46), 13q (7/46), 17p (12/46), 18q (7/46), Xp (18/46) and Xq (19/46), with smallest overlapping regions at 2q32-qter (16/46), 5q12-q31 (8/46), 8p12-pter (7/46), 9p21-pter (10/46), 9q (13/46), 11p (7/46), 13q13-q22 (7/46), 17p (12/46), 18q21-qter (7/46), Xp (18/46) and Xq (19/46). There were significantly higher frequencies of gains of 1q21-q24 and 17q22-qter in moderately differentiated tumors as compared with those in well-differentiated tumors, indicating a possible association of these two abnormalities with the dedifferentiation of tumor cells. Gains of 1p32-pter, 5p, 6p22-p23, 11q13-q14, 17q22-qter and losses of 2q32-qter, 9q, 17p were more frequent in pT1 as compared with those in pTa carcinomas. Gains at 1q21-q24, 7p11.2-p14, 8q22-q24, 19q, 20q11-q13.2 and losses at 5q12-q31, 8p12-pter, 9p21-pter, 11p, 13q13-q22 and 18q21-qter were unique to pT1 and higher stage tumors, suggesting that genes responsible for the invasion and progression of bladder cancer might be located at these chromosomal regions. In multiple tumors from the same patients, consistent alterations such as gains of 8q, 11q13-q14, 12q13-q15, 13q12, 20q and losses of 2q32-qter, 8p, 9, 11p, 11q21-qter, 13q13-qter, X were detected. These abnormalities were possibly earlier events, which might play a critical role during the genesis of the tumors. Further detailed studies to the recurrent aberration regions may lead to the identification of oncogenes and tumor suppressor genes involved in the development and progression of Chinese bladder cancer.