Bladder cancer is one of the most common malignancies of the urinary tract. Among its subtypes, muscle invasive bladder cancer (MIBC) is particularly aggressive and often associated with poor prognosis. The efficacy of platinum-based neoadjuvant chemotherapy and radical cystectomy remains unsatisfactory. In recent years, immune checkpoint inhibitors have shown promising therapeutic potential in MIBC. However, reliable biomarkers for predicting treatment response are still lacking. Moreover, traditional clinical parameters, such as TNM staging, often fail to accurately assess patient outcomes. Therefore, identifying novel biomarkers is crucial for improving prognosis and optimizing treatment strategies in MIBC. We first analyzed the expression pattern and prognostic significance of methylenetetrahydrofolate dehydrogenase (NADP + dependent) 1-like (MTHFD1L) across multiple cancer types. Subsequently, MIBC patient samples from three independent cohorts (TCGA-BLCA, GSE169455, and GSE48075) were used to evaluate the prognostic value of MTHFD1L through univariate and multivariate Cox regression analyses combined with Kaplan-Meier survival analysis. Gene Set Enrichment Analysis and Gene Set Variation Analysis were performed to explore the biological functions of MTHFD1L. Mutation characteristics and immunotherapy response-related features were further analyzed based on MTHFD1L expression levels. Finally, the results were further verified by immunohistochemistry and cell function experiments. MTHFD1L was upregulated in multiple cancer types, and its elevated expression was significantly associated with unfavorable outcomes. In all independent MIBC cohorts, high MTHFD1L expression served as an independent risk factor for poor prognosis. Gene Set Enrichment Analysis indicated that MTHFD1L may promote tumor progression by activating immune-related and proliferative pathways while suppressing metabolic processes. Mutation analysis revealed a higher frequency of TP53 mutations in the MTHFD1L high-expression group. Moreover, immune response prediction suggested that patients with low MTHFD1L expression were more likely to benefit from immune checkpoint inhibitors therapy. Immunohistochemistry confirmed the overexpression of MTHFD1L in MIBC tissues, and cell function experiments demonstrated that MTHFD1L knockdown markedly inhibited bladder cancer cell proliferation, colony formation, and migration. MTHFD1L represents a promising and reliable biomarker for predicting prognosis and immunotherapy response in MIBC, providing a new foundation for the development of precision and personalized therapeutic strategies.

The poor prognosis of bladder cancer (BCa) is primarily attributed to the acquisition of invasive and metastatic capabilities by tumor cells through epithelial-mesenchymal transition (EMT), yet the dynamic alterations in ferroptosis during EMT and their regulatory mechanisms remain unelucidated. This study is the first to reveal the mechanism by which the Smad3/CISD2 signaling axis regulates ferroptosis in mesenchymal-like bladder cancer cells. Clinical sample analysis demonstrated significantly reduced expression of E-cadherin and upregulation of N-cadherin and Vimentin in muscle-invasive bladder cancer tissues (MIBC), with EMT-related marker levels correlating with overall survival rates in BCa patients. In TGF-β1-induced mesenchymal-like bladder cancer cells, ferroptosis-related genes (GPX4, SLC7A11) were markedly elevated, alongside increased lipid peroxides (LPO) and glutathione (GSH) levels. However, mesenchymal-like bladder cancer cells exhibited heightened sensitivity to the ferroptosis inducer Erastin, showing more pronounced suppression of proliferation, elevated ROS, higher LPO and MDA levels, and reduced GSH, confirming their enhanced susceptibility to ferroptosis. RNA-seq revealed significant downregulation of Smad3 in ferroptosis inducer treated mesenchymal-like bladder cancer cells. Smad3 knockdown further exacerbated ferroptosis markers (elevated Fe²⁺, ROS, LPO; decreased GSH), inhibited migration, invasion, and proliferation—phenotypes reversible by ferroptosis inhibitors. Mechanistically, ChIP-seq combined with RNA-seq demonstrated that Smad3 regulates CISD2 expression by binding its promoter region, with clinical specimens confirming a positive correlation between Smad3 and CISD2 expression. Functional rescue experiments showed that CISD2 overexpression in Smad3-knockdown mesenchymal-like bladder cancer cells reversed abnormal increases in Fe²⁺, ROS, LPO, and MDA while restoring GSH levels, indicating that Smad3 modulates ferroptosis through a CISD2-dependent pathway. In vivo experiments further demonstrated that targeting the Smad3/CISD2 axis significantly suppressed xenograft tumor growth and activated ferroptosis. In conclusion, this study elucidates a novel mechanism by which the Smad3/CISD2 axis dynamically regulates ferroptosis through redox homeostasis reprogramming during EMT, providing a potential therapeutic strategy for targeting the progression of MIBC.

Muscle‐invasive bladder cancer (MIBC) poses a severe threat to patient survival due to its high invasiveness and metastatic potential. Although immunotherapy has revolutionized treatment strategies for MIBC, immune evasion remains a major obstacle limiting therapeutic efficacy. In this study, the mitochondrial calcium uniporter regulatory subunit (MCUB) is investigated for its role in immune evasion in MIBC. Bulk RNA‐seq, scRNA‐seq, and proteomic analyses revealed a progressive upregulation of MCUB from normal to MIBC tissues, and strong positive correlations are uncovered between MCUB expression and both PD‐L1/PD‐1 signaling and poor outcomes. Spatial transcriptomics and clinical tissue staining confirmed spatial co‐localization of MCUB and PD‐L1. Functional experiments demonstrated that MCUB stabilized PD‐L1 protein by reducing its lysosomal degradation through inhibition of PRKN‐dependent mitophagy. Mechanistically, MCUB suppressed mitochondrial calcium uptake to reduce PRKN activation and physically interacted with the PRKN‐Arg51 residue to inhibit its function. In vivo, MCUB knockdown led to reduced tumor growth, enhanced CD8⁺ T cell infiltration, and improved response to anti‐PD‐1 therapy. This study identified the MCUB‐PRKN‐PD‐L1 axis as a novel driver of immune evasion in MIBC and proposed that targeting the MCUB‐PRKN interaction may serve as a precise therapeutic strategy to overcome immune resistance with minimal toxicity to normal tissues.

Bladder cancer (BLCA) is the second most common urologic cancer in the United States and worldwide and mostly affects the aging population. Despite several ongoing clinical trials, treatment paradigms for BLCA have not changed significantly. Here, we investigated the expression of transient receptor potential vanilloid type 4 (TRPV4) in patients with BLCA and its role in calcium influx, cell proliferation, and migration using normal human urothelial cells and BLCA cells. Bioinformatic analysis of the University of Alabama at Birmingham Cancer Data Analysis Portal and cBioPortal databases revealed that TRPV4 expression is significantly higher in human BLCA tissues than in normal adjacent tissues. Furthermore, TRPV4 expression was markedly elevated in early-stage BLCA and upregulated in muscle-invasive bladder cancer tissues. TRPV4 is expressed in both normal urothelial (SV-HUC-1) and BLCA (T-24) cells, and functional assays demonstrated enhanced TRPV4-mediated calcium influx in T-24 compared with SV-HUC-1 cells. T-24 cells exhibited higher spreading on extracellular matrix gels with increasing stiffness (0.2, 8, and 50 kPa) and exhibited a migratory phenotype compared to SV-HUC-1 cells. Pharmacological inhibition of TRPV4 significantly reduced proliferation and migration in T-24 cells but had minimal effects on normal cells. Finally, treatment with cisplatin significantly reduced TRPV4 protein levels and TRPV4-mediated calcium influx in chemosensitive UM-UC-3 cells but remained unchanged in chemoresistant T-24 cells, suggesting a potential role of TRPV4 in chemoresistance. In conclusion, TRPV4 may contribute to BLCA progression by regulating cell proliferation and migration and may impart resistance to chemotherapy. Targeting TRPV4 could present a novel therapeutic approach for managing BLCA progression and overcoming chemoresistance.Significance StatementThis study identified transient receptor potential vanilloid type 4 (TRPV4) as a critical driver of bladder cancer (BLCA) progression. TRPV4 gene expression is elevated in both early-stage and muscle-invasive BLCA tissues. Importantly, TRPV4 inhibition selectively reduces BLCA growth and motility. Furthermore, TRPV4 is downregulated by cisplatin in chemosensitive but not chemoresistant BLCA cells, underscoring its key role in bladder cancer chemoresistance. These findings position TRPV4 as a therapeutic target for enhancing BLCA treatment and overcoming drug resistance.

BackgroundHow to select muscle-invasive bladder cancer (MIBC) patients who are sensitive to immunotherapy is an unmet medical need. This study aimed to explore the role of immunoproteasome subunits as a novel signature for predicting efficacy of immunotherapy in MIBC.MethodsThe expression profile of immunoproteasome subunits of MIBC and normal tissues was evaluated from data of The Cancer Genome Atlas (TCGA) and of the Chongqing University Cancer Hospital (CQUCH) cohort. Survival analysis and response to immunotherapy was further explored and compared between immunoproteasome subunitshigh and immunoproteasome subunitslow MIBC patients in the TCGA, the CQUCH and the IMvigor210 cohort. The association of the expression of immunoproteasome subunits with immune checkpoint molecules and the tumor immune microenvironment was explored by immunohistochemistry staining and bioinformatic analysis in MIBC of these three cohorts.ResultsThe expression of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 was significantly upregulated in MIBC. MIBC patients with high expression of immunoproteasome subunits, especially high expression of PSMB9, showed a trend of prolonged overall and progression free survival, which was further significantly improved in response to immunotherapy. Bioinformatics and immunohistochemistry staining revealed a positive correlation of the expression of immunoproteasome subunits with the expression of immune checkpoint molecules, with T cell activation and with T cell-mediated cytotoxicity.ConclusionsImmunoproteasome subunits, in particular PSMB9, are immune microenvironment-related molecules of MIBC and are promising signatures for survival prediction in response to immunotherapy of MIBC.Graphical

Bladder cancer (BC) is a prevalent urinary malignancy and muscle-invasive bladder cancer (MIBC) is particularly aggressive and associated with poor prognosis. One of MIBC features is the nuclear atypia. However, the molecular mechanism underlying MIBC remains unclear. Here, we find that FKBP10 is significantly upregulated in MIBC tissues and correlated with metastasis and poor outcomes. FKBP10 promotes tumor cell invasion, migration, and metastasis, but not proliferation. Notably, FKBP10 enhances the nuclear atypia of BC cells. Mechanistically, FKBP10 interacts with prelamin A and hinder the nuclear entry of prelamin A, thereby leading to the decrease in the nuclear lamin A, a key factor involved in nuclear atypia. In human BC tissues, nuclear lamin A is downregulated and negatively correlated with FKBP10 expression. Overall, our findings demonstrate that the FKBP10/prelamin A/lamin A axis contributes to MIBC.

This study investigates the potential of purine nucleoside phosphorylase (PNP) as a biomarker and therapeutic target in muscle-invasive bladder cancer (MIBC). We aimed to explore PNP's expression, prognostic value, and role in metabolic pathways, along with its association with gene mutations. We conducted multi-omics analyses using data from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), and other public databases to evaluate PNP expression across MIBC samples and its prognostic impact through Kaplan-Meier and Cox regression analyses. Functional enrichment and gene set variation analysis (GSVA) were performed to identify PNP-related pathways. In addition, in vitro siRNA knockdown experiments were carried out to assess PNP's influence on MIBC cell proliferation. Our findings revealed that PNP is significantly overexpressed in MIBC tissues and serves as an independent prognostic factor, correlating with poor clinical outcomes across multiple cohorts (TCGA: hazard ratio [HR] > 1.3, P < .05; GSE48075: HR > 1.5, P = .07; GSE169455: HR > 2.8, P < .001). Functional enrichment analysis identified PNP's involvement in various metabolic pathways. Furthermore, we observed a high frequency of RB1 mutations in the PNP-high expression group. Based on this observation, we hypothesize that patients harboring RB1 mutations may benefit from PNP-targeted therapy. In vitro experiments demonstrated that PNP knockdown significantly reduces MIBC cell proliferation. This study underscores PNP's role as a promising biomarker and therapeutic target in MIBC.

PurposeUrothelial carcinoma has various molecular subtypes, each with different tumor characteristics. Although it is known that molecular changes occur during tumor progression, little is known about the specifics of these changes. In this study, we performed transcriptional analysis to understand the molecular changes during tumor progression.Materials and MethodsFormalin-fixed, paraffin-embedded tumor tissues were obtained from 12 patients with muscle-invasive bladder cancer (MIBC). The invasive and non-invasive papillary areas were identified in papillary urothelial carcinoma specimens. Immunohistochemistry (IHC) and mRNA sequencing were performed for each tumor area.ResultsPatients with CK5/6-negative and CK20-positive non-invasive papillary areas were selected and classified into the IHC switch subgroup (CK5/6-positive and CK20-negative in the invasive area) and the IHC unchanged subgroup (CK5/6-negative and CK20-positive in the invasive area) according to the IHC results of the invasive area. We identified differences in the mRNA expression between the non-invasive papillary and invasive areas of the papillary MIBC tissue samples. In both the non-invasive papillary and invasive areas, the IHC switch subgroup showed basal subtype gene expression, while the IHC unchanged subgroup demonstrated luminal subtype gene expression.ConclusionsThe non-invasive papillary area showed a gene expression pattern similar to that of the invasive area. Therefore, even if the non-invasive papillary area exhibits a luminal phenotype on IHC, it can have a basal subtype gene expression depending on the invasive area.

Bladder cancer (BCa) is the most common malignancy of the urinary tract which can be divided into non-muscle-invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC), and their microbial differences are not fully understood. This study was conducted by performing 2bRAD sequencing for Microbiome (2bRAD-M) on NMIBC and MIBC tissue samples to investigate the microbiota differences between NMIBC and MIBC individuals. A total of 22 patients with BCa, including 7 NMIBC and 15 MIBC, were recruited. Tumor tissues were surgically removed as samples and DNA was extracted. Type IIB restriction endonucleases were used to enzymatically cleave the microbial genome for each microbe's tag and map it to species-specific 2bRAD markers to enable qualitative and quantitative studies of microbes between MIBC and NMIBC tissues. A total of 527 species were detected. The microbial diversity of NMIBC tissues was significantly higher than that of MIBC tissues. Microbial composition of the two tumor tissues was similar, where Ralstonia_sp000620465 was the most dominant species. 4 species (Acinetobacter_guillouiae, Anoxybacillus_A_rupiensis, Brevibacillus_agri and Staphylococcus_lugdunensis) were enriched in NMIBC, while Ralstonia_mannitolilytica, Ralstonia_pickettii, and Ralstonia_sp000620465 were overrepresented in MIBC. 252 discriminatory character taxa were also revealed by linear discriminant analysis effect sizea (LEfSe). Species importance point plots identified Ralstonia_sp000620465, Cutibacterium_acnes and Ralstonia_pickettii as the three most important species between the two groups. Meanwhile, functional annotation analysis showed 3011 different COGs and 344 related signaling pathways between MIBC and NMIBC microbiome. This first 2bRAD-M microbiome study on MIBC and NMIBC tissues revealed significant differences in the microbial environment between the two groups, which implies a potential association between tumor microbial dysbiosis and BCa, and provides a possible target and basis for subsequent studies on the mechanisms of BCa development and progression.

Muscle-invasive bladder cancer (MIBC) is characterized as bladder tumors that infiltrate into the muscle layer, along with multiple metastasis and poor prognosis. Numerous research studies have been performed to identify the underlying clinical and pathological alterations that occur. However, few studies have revealed the molecular mechanism of its progression based upon the immunotherapy response. Our present study was designed to identify biomarkers which may predict the immunotherapy response by investigating the tumor microenvironment (TME) in MIBC. The transcriptome and clinical data of MIBC patients were obtained and analyzed with R version 4.0.3 (POSIT Software, Boston, MA, USA) ESTIMATE package. Differentially expressed immune-related genes (DEIRGs) were identified and further analyzed via the protein-protein interaction network (PPI). Meanwhile, univariate Cox analysis was utilized to screen out the prognostic DEIRGs (PDEIRGs). Then, the PPI core gene was matched with PDEIRGs to obtain the target gene-fibronectin-1 (FN1). Human MIBC and control tissues were collected and FN1 was measured with Quantitative Reverse Transcription PCR (qRT-PCR) and Western-Blot. Finally, the relationship between FN1 expression level and MIBC was validated through survival, univariate Cox, multivariate Cox, Gene Set Enrichment Analysis (GSEA) and correlation analysis of tumor infiltrating immune cells. TME DEIRGs were identified and the target gene FN1 was obtained. The higher expression of FN1 was confirmed in MIBC tissues via bioinformatics analysis, qRT-PCR and Western-Blot. Moreover, higher FN1 expression correlated with reduced survival time and FN1 expression was further favorably correlated with clinic-pathological features (grade, TNM stage, invasion, lymphatic and distant metastasis). Additionally, the genes in the high FN1 expression group were mainly enriched in immune-related activities and macrophage M2, T cell CD4, T cell CD8 and T cell follicular helper cells were correlated with FN1. Finally, it was observed that FN1 was closely related to key immune checkpoints. FN1 was identified as a novel and independent prognostic factor for MIBC. Our data also suggests FN1 can predict MIBC patients' response to immune checkpoints inhibitors.

Methylene blue (MB) is a dye used for histology with clinical importance and intercalates into nucleic acids. After MB staining of formalin fixed paraffin embedded (FFPE) muscle invasive bladder cancer (MIBC) and normal urothelium, specific regions could be microdissected. It is not known if MB influences RNA used for gene expression studies. Therefore, we analyzed MIBC using five different RNA isolation methods comparing patient matched FFPE and fresh frozen (FF) tissues pre-stained with or without MB. We demonstrate a positive impact of MB on RNA integrity with FF tissues using real time PCR with no interference of its chemical properties. FFPE tissues showed no improvement of RNA integrity, which we propose is due to formalin induced nucleotide crosslinks. Using direct multiplex RNA hybridization the best genes for normalization of MIBC and control tissues were identified from 34 reference genes. In addition, 5SrRNA and 5.8SrRNA were distinctive reference genes detecting <200 bp fragments important for mRNA analyses. Using these normalized RNAs from MB stained MIBC and applying multiplex RNA hybridization and mRNA sequencing, a minimal gene expression panel precisely identified luminal and basal MIBC tumor subtypes, important for diagnosis, prognosis and chemotherapy response.

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). 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). 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. 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.

Bladder cancer (BCa) is a common cancer associated with high morbidity and mortality worldwide. Pre-B-cell leukemia transcription factor 1 (PBX1) has been reported to be involved in tumor progression. The aim of the study was to explore the specific role of PBX1 in BCa and its underlying mechanisms. The relative expressions of PBX1 in muscle-invasive BCa tissues and cell lines were analyzed through RT-qPCR and western blotting. Kaplan-Meier analysis was used to analyze the relationship between PBX1 levels and survival status. Co-immunoprecipitation (CO-IP) and chromatin immunoprecipitation (ChIP)-qPCR assays were adopted to verify the interaction between PBX1 and Estrogen receptors (ERs) and explore the estrogen receptors (ERs)-dependent genes transcription. PBX1 was upregulated in invasive BCa patients and BCa cells, positively associated with tumor size, lymph node metastasis, distant metastasis and poorer survival status. The overexpression of PBX1 promoted cell growth, invasion, epithelial-mesenchymal transition (EMT) process and cisplatin resistance in BCa cells, while the silence of PBX1 showed opposite effects. Furthermore, PBX1 interacted with ERs and was required for ER function. PBX1 overexpression aggravated the tumorpromoting effect of estrogen on BCa cells, while it partially suppressed the inhibitory effects of ER antagonist AZD9496 on BCa cells. This study revealed that PBX1 participated in estrogen mediated BCa progression and chemo-resistance through binding and activating estrogen receptors. Hence, PBX1 may serve as a potential prognostic and therapeutic target for BCa treatment.

Chemotherapy is a standard therapy for muscle-invasive bladder cancer (MIBC). However, genomic alterations associated with chemotherapy sensitivity in MIBC have not been fully explored. This study aimed to investigate the genomic landscape of MIBC in association with the response to chemotherapy and to explore the biological role of genomic alterations. Genomic alterations in MIBC were sequenced by targeted exome sequencing of 409 genes. Gene expression in MIBC tissues was analyzed by western blotting, immunohistochemistry, and RNA microarray. Cellular sensitivity to gemcitabine and gemcitabine metabolite was examined in bladder cancer cells after modulation of candidate gene. Targeted exome sequencing in 20 cases with MIBC revealed various genomic alterations including pathogenic missense mutation of DPYD gene encoding dihydropyrimidine dehydrogenase (DPD). Conversely, high DPYD and DPD expression were associated with poor response to gemcitabine-containing chemotherapy among patients with MIBC, as well as gemcitabine resistance in bladder cancer cells. DPD suppression rendered cells sensitive to gemcitabine, while DPD overexpression made cells gemcitabine-resistant through reduced activity of the cytotoxic gemcitabine metabolite difluorodeoxycytidine diphosphate. This study revealed the novel role of DPD in gemcitabine metabolism. It has been suggested that DPYD genomic alterations and DPD expression are potential predictive biomarkers in gemcitabine treatment.

Muscle-invasive bladder cancer (MIBC) leads to a large societal burden. Recently, tRNA-derived small RNAs (tsRNAs), a novel type of noncoding RNA (ncRNAs), have been identified. However, the expression patterns and functions of tsRNAs in MIBC have not yet been identified. Here, RNA sequencing, bioinformatics, and quantitative reverse transcription- polymerase chain reaction (qRT-PCR) were used to screen the expression profiles and predict the potential roles of tsRNAs in MIBC. Of 406 tsRNAs differentially expressed in MIBC tissues, 91 tsRNAs were significantly differentially expressed. Then, four candidate tsRNAs, tiRNA-1:34-Val-CAC-2, tiRNA-1:33-Gly-GCC-1, tRF-1:32-Gly-GCC-1, and tRF-+1:T20-Ser-TGA-1, were selected. Next, a bioinformatics analysis showed the potential target genes and tsRNA–mRNA network. The most significant and meaningful terms of gene ontology were the positive regulation of the phosphate metabolic process, lamellipodium, and protein-cysteine S-acyltransferase activity in the biological process, cellular component, and molecular function, respectively. In addition, the top four pathways were predicted by the Kyoto Encyclopedia of Genes and Genomes database (KEGG). Finally, qRT-PCR demonstrated a similar expression pattern compared to sequencing data for the candidate tsRNAs. In short, we find differential expression profiles and predict that tiRNA-1:33-Gly-GCC-1, tRF-1:32-Gly-GCC-1, and tRF-+1:T20-Ser-TGA-1 are very likely to engage in the pathophysiological process of MIBC via regulating the target genes in the key pathways.

Simple SummaryAssessing the tumor response to chemotherapy is a paramount predictive step to improve patient care. Infrared spectroscopy probes the chemical composition of samples, and in combination with statistical multivariate processing, presents the capacity to highlight subtle molecular alterations associated with malignancy characteristics. Microscopic infrared imaging of tissue samples reveals spectral heterogeneity within histological structures, providing a new approach to characterize tumoral heterogeneity. We have taken advantage of the analytical capabilities of mid-infrared spectral imaging to implement a classification model to predict the response of a tumor to chemotherapy. Our development was demonstrated in muscle-invasive bladder cancer (MIBC) by comparing samples from responders and non-responders to neoadjuvant chemotherapy.Background: Neoadjuvant chemotherapy (NAC) improves survival in responder patients. However, for non-responders, the treatment represents an ineffective exposure to chemotherapy and its potential adverse events. Predicting the response to treatment is a major issue in the therapeutic management of patients, particularly for patients with muscle-invasive bladder cancer. Methods: Tissue samples of trans-urethral resection of bladder tumor collected at the diagnosis time, were analyzed by mid-infrared imaging. A sequence of spectral data processing was implemented for automatic recognition of informative pixels and scoring each pixel according to a continuous scale (from 0 to 10) associated with the response to NAC. The ground truth status of the responder or non-responder was based on histopathological examination of the samples. Results: Although the TMA spots of tumors appeared histologically homogeneous, the infrared approach highlighted spectral heterogeneity. Both the quantification of this heterogeneity and the scoring of the NAC response at the pixel level were used to construct sensitivity and specificity maps from which decision criteria can be extracted to classify cancerous samples. Conclusions: This proof-of-concept appears as the first to evaluate the potential of the mid-infrared approach for the prediction of response to neoadjuvant chemotherapy in MIBC tissues.

Receptor Activator of NF Kappa B (RANK) Expression Indicates Favorable Prognosis in Patients with Muscle-invasive Bladder Cancer

IntroductionBladder cancer ranks the first in the morbidity of urogenital malignancies in China. Bladder cancers are pathologically classified into 2 sub-types, including non-muscle invasion bladder cancer (NMIBC) and muscle invasive bladder cancer (MIBC). MIBC is a highly lethal tumor and targeted therapies showed promising prospect for the treatment of MIBC. Novel therapeutic targets are still badly needed to combat this disease. The kinesin family member 14 (KIF14) is an engaging molecular motor and involved in multiple cellular processes such as cell division. Additionally, KIF14 is highly expressed in multiple tumor tissues and participates in the progression of several cancers such as gastric cancer and hepatocellular carcinoma. However, its possible role in the development of bladder cancer remains unclear.Material and methodsHerein, 107 cases of MIBC tissue specimens were collected and detected by immunohistochemistry assays, and we analyzed the relationship between AKIF14 expression and clinical features. Then we used the cell line T24 and 5637 of bladder cancer into the experimental group transfected shKIF14 plasmid.KIF14, additionally, fascinated tumor growth of MIBC in mice.ResultsWe demonstrated the high expression of KIF14 in tumor tissues from patients who underwent MIBC. Furthermore, KIF14 was statistically correlated with clinical feature, such as tumor stage (P = 0.001). Our results further confirmed the impairment of proliferation capacity after KIF14 ablation in vitro and in vivo.ConclusionsIn summary, we revealed KIF14 could serve as a promising therapeutic target for the treatment of MIBC.

Simple SummaryMuscle-invasive bladder cancer (MIBC) accounts for the majority of bladder cancer mortality worldwide. Clinical assessment of MIBC mainly relies on the TNM staging system to provide guidance for both prognosis and therapy planning. Based on standardized quantification of tumour-immune features across whole slide images, and in conjunction with clinical information, we construct an ensemble machine learning model that correctly classifies 71.4% of the patients who succumb to MIBC, significantly higher than the 28.6% of TNM staging system. Post-hoc analysis of our model reveals clinically relevant, immunological features for MIBC prognosis, thereby further supporting their adoption into the clinic.The clinical staging and prognosis of muscle-invasive bladder cancer (MIBC) routinely includes the assessment of patient tissue samples by a pathologist. Recent studies corroborate the importance of image analysis in identifying and quantifying immunological markers from tissue samples that can provide further insight into patient prognosis. In this paper, we apply multiplex immunofluorescence to MIBC tissue sections to capture whole-slide images and quantify potential prognostic markers related to lymphocytes, macrophages, tumour buds, and PD-L1. We propose a machine-learning-based approach for the prediction of 5 year prognosis with different combinations of image, clinical, and spatial features. An ensemble model comprising several functionally different models successfully stratifies MIBC patients into two risk groups with high statistical significance (p value < ). Critical to improving MIBC survival rates, our method correctly classifies of the patients who succumb to MIBC, which is significantly more than the of the current clinical gold standard, the TNM staging system.

Muscle-invasive bladder cancer (MIBC) is characterized by a highly complex immune environment, which is not well understood. Interleukin-6 (IL-6) is generated and secreted by multifarious types of cells, including tumor cells. This study was aimed at demonstrating that the levels of IL-6 and the number of myeloid-derived suppressor cells (MDSCs), with a positive correlation between them, increased in MIBC tissues, promoting MIBC cell proliferation, especially in patients with recurrence. In coculture analysis, MDSCs, with the stimulation of IL-6, could significantly lower the proliferation ability of CD4+ or CD8+ T lymphocytes. Further, this study demonstrated that IL-6 could upregulate the mitogen-activated protein kinase (MAPK) signaling pathway in MDSCs. The MAPK signaling inhibitor, aloesin, partially reversed the effects of IL-6 on MDSCs. These data suggested that IL-6 promoted MIBC progression by not only accelerating proliferation but also improving the immune suppression ability of MDSCs through activating the MAPK signaling pathway.