Bladder Cells Research Articles

Epidermal bladder cells play a role in water retention in quinoa leaves

Epidermal bladder cells play a role in water retention in quinoa leaves

Mechanobiology studies of bladder tumor cells using laterally squeezing microfluidic flow cytometry

The deformability and uptake capability of cells are critical indicators of their biomechanical properties and functional behaviors, particularly in tumor heterogeneity and cancer research. Here, we introduce a microfluidic flow cytometry platform integrated with a laterally adjustable squeezing structure for the characterization of bladder tumor cells (including 5637 and EJ cell lines) and uroepithelial cells (SV-HUC-1 cell line). The deformability of these cell types under varying channel width conditions was clearly assessed using this platform. The results demonstrated that tumor cells exhibited higher deformability compared to uroepithelial cells, with the EJ cell line exhibiting the greatest difference. Furthermore, the relationship between the malignancy, deformability, and uptake capability of bladder cells was explored through co-cultivation experiments with 2 μm particles. As the malignancy increased, the cells became more deformable and exhibited stronger phagocytic capability with particles. Subsequently, the heterogeneity of tumor cells was investigated by analyzing the deformability of phagocytic and non-phagocytic subpopulations within EJ cells. The developed microfluidic platform offers a promising high-throughput method to assess the biomechanical and phagocytic characteristics of cells, providing valuable insights into tumor cell biology, and potentially improving clinical status of urinary cytology examinations for bladder cancer.

119 - Modulating cyclic nucleotides reduces stress-induced ATP release from isolated urothelial cells of the mouse bladder

119 - Modulating cyclic nucleotides reduces stress-induced ATP release from isolated urothelial cells of the mouse bladder

Combination screen in multi-cell type tumor spheroids reveals interaction between aryl hydrocarbon receptor antagonists and E1 ubiquitin-activating enzyme inhibitor

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates genes of drug transporters and metabolic enzymes to detoxify small molecule xenobiotics. It has a complex role in cancer biology, influencing both the progression and suppression of tumors by modulating malignant properties of tumor cells and anti-tumor immunity, depending on the specific tumor type and developmental stage. This has led to the discovery and development of selective AhR modulators, including BAY 2416964 which is currently in clinical trials. To identify small molecule anticancer agents that might be combined with AhR antagonists for cancer therapy, a high-throughput combination screen was performed using multi-cell type tumor spheroids grown from malignant cells, endothelial cells, and mesenchymal stem cells. The AhR selective antagonists BAY 2416964, GNF351, and CH-223191 were tested individually and in combination with twenty-five small molecule anticancer agents. As single agents, BAY 2416964 and CH-223191 showed minimal activity, whereas GNF351 reduced the viability of some spheroid models at concentrations greater than 1 µM. The activity of most combinations aligned well with the single agent activity of the combined agent, without apparent contributions from the AhR antagonist. All three AhR antagonists sensitized tumor spheroids to TAK-243, an E1 ubiquitin-activating enzyme inhibitor. These combinations were active in spheroids containing bladder, breast, ovary, kidney, pancreas, colon, and lung tumor cell lines. The AhR antagonists also potentiated pevonedistat, a selective inhibitor of the NEDD8-activating enzyme E1 regulatory subunit, in several tumor spheroid models. In contrast, the AhR antagonists did not enhance the cytotoxicity of the proteasome inhibitor bortezomib.

Concurrent Urothelial Carcinoma of the Bladder and Renal Cell Carcinoma of the Kidney: A Diagnostic and Therapeutic Challenge

Concurrent Urothelial Carcinoma of the Bladder and Renal Cell Carcinoma of the Kidney: A Diagnostic and Therapeutic Challenge

Genetic requirements for uropathogenic E. coli proliferation in the bladder cell infection cycle.

Urinary tract infections (UTIs) are one of the most frequent infections worldwide. Uropathogenic Escherichia coli (UPEC), which accounts for ~80% of UTIs, must rapidly adapt to highly variable host environments, such as the gut, bladder sub-surface, and urine. In this study, we searched for UPEC genes required for bacterial growth and survival throughout the cellular infection cycle. Genes required for de novo synthesis of biomolecules and cell envelope integrity appeared to be important, and other genes were also implicated in bacterial dispersal and recovery from infection of cultured bladder cells. With further studies of individual gene function, their potential as therapeutic targets may be realized. This study expands knowledge of the UTI cycle and establishes an approach to genome-wide functional analyses of stage-resolved microbial infections.

Pharmacological Inhibition of Endogenous Hydrogen Sulfide Production Slows Bladder Cancer Progression in an Intravesical Murine Model.

Present bladder cancer therapies have relatively limited therapeutic impact and account for one of the highest lifetime treatment costs per patient. Therefore, there is an urgent need to explore novel and optimized treatment strategies. The present study investigated the effects of inhibiting endogenous hydrogen sulfide (H2S) production on bladder cell viability and in vivo tumor progression. We targeted the H2S-producing enzyme, cystathionine γ-lyase, in 5637 cells using propargylglycine (H2S inhibitor) and performed cytofluorimetric analysis to evaluate cell viability. We then tested the efficacy of propargylglycine alone or in combination with gemcitabine (conventional chemotherapy) in an intravesical murine model of bladder cancer. Magnetic resonance imaging and immunohistochemical staining for cell proliferation, apoptosis, immune-cell infiltration, and neovascularization were performed to evaluate tumor response. Compared to control conditions or cohorts, propargylglycine administration significantly attenuated bladder cancer cell viability in vitro (p < 0.0001) and tumor growth (p < 0.002) and invasion in vivo. Furthermore, propargylglycine enhanced the anti-cancer effects of gemcitabine, resulting in tumor regression (p < 0.0001). Moreover, propargylglycine induced cleaved PARP-1-activated apoptosis (p < 0.05), as well as intratumoral CD8+ T cell (p < 0.05) and F4/80+ macrophage (p < 0.002) infiltration. Propargylglycine also reduced intratumoral neovascularization (p < 0.0001) and cell proliferation (p < 0.0002). Importantly, the pro-apoptotic and anti-neovascularization effects of gemcitabine were enhanced by propargylglycine co-administration. Our findings suggest that inhibition of endogenous H2S production can be protective against bladder cancer by enhancing the chemotherapeutic action of gemcitabine and may be a novel pharmacological target and approach for improved bladder cancer diagnosis and treatments in the future.

Contribution of transient receptor potential vanilloid 1 (TRPV1) channel to cholinergic contraction of rat bladder smooth muscle.

Transient receptor potential vanilloid 1 (TRPV1) is a calcium-permeable ion channel that is gated by the pungent constituent of red chili pepper, capsaicin, and by related chemicals from the group of vanilloids, in addition to noxious heat. It is expressed mostly in sensory neurons to act as a detector of painful stimuli produced by pungent chemicals and high temperatures. Although TRPV1 is also found outside the sensory nervous system, its expression and function in the bladder detrusor smooth muscle (DSM) remain controversial. Here, by using Ca2+ imaging and patch clamp on isolated rat DSM cells, in addition to tensiometry on multicellular DSM strips, we show that TRPV1 is expressed functionally in only a fraction of DSM cells, in which it acts as an endoplasmic reticulum Ca2+-release channel responsible for the capsaicin-activated [Ca2+]i rise. Carbachol-stimulated contractions of multicellular DSM strips contain a TRPV1-dependent component, which is negligible in the circular DSM but reaches ≤50% in the longitudinal DSM. Activation of TRPV1 in rat DSM during muscarinic cholinergic stimulation is ensured by phospholipase A2-catalysed derivation of arachidonic acid and its conversion by lipoxygenases to eicosanoids, which act as endogenous TRPV1 agonists. Immunofluorescence detection of TRPV1 protein in bladder sections and isolated DSM cells confirmed both its preferential expression in the longitudinal DSM sublayer and its targeting to the endoplasmic reticulum. We conclude that TRPV1 is an essential contributor to the cholinergic contraction of bladder longitudinal DSM, which might be important for producing spatial and/or temporal anisotropy of bladder wall deformation in different regions during parasympathetic stimulation. KEY POINTS: The transient receptor potential vanilloid 1 (TRPV1) heat/capsaicin receptor/channel is localized in the endoplasmic reticulum membrane of detrusor smooth muscle (DSM) cells of the rat bladder, operating as a calcium-release channel. Isolated DSM cells are separated into two nearly equal groups, within which the cells either show or do not show TRPV1-dependent [Ca2+]i rise. Carbachol-stimulated, muscarinic ACh receptor-mediated contractions of multicellular DSM strips contain a TRPV1-dependent component. This component is negligible in the circular DSM but reaches ≤50% in longitudinal DSM. Activation of TRPV1 in rat DSM during cholinergic stimulation involves phospholipase A2-catalysed derivation of arachidonic acid and its conversion by lipoxygenases to eicosanoids, which act as endogenous TRPV1 agonists.

Identification and Validation of Prognostic Model for Tumor Microenvironment-Associated Genes in Bladder Cancer Based on Single-Cell RNA Sequencing Data Sets.

The purpose of this study was to elucidate the relationship between the tumor microenvironment (TME) and cellular diversity in bladder cancer (BLCA) progression, leveraging single-cell RNA sequencing (scRNA-seq) data to identify potential prognostic biomarkers and construct a prognostic model for BLCA. We analyzed scRNA-seq data of normal and tumor bladder cells from the Gene Expression Omnibus (GEO) database to uncover crucial markers within the bladder TME. The study compared gene expression in normal versus tumor bladder cells, identifying differentially expressed genes. These genes were subsequently assessed for their prognostic significance using patient follow-up data from The Cancer Genome Atlas. Prognostic models were constructed using Least Absolute Shrinkage and Selection Operator and multivariate Cox regression analyses, focusing on eight genes of interest. The predictive performance of the model was also tested against additional GEO data sets (GSE31684, GSE13507, and GSE32894). The prognostic model demonstrated reliable prediction of patient outcomes. Validation through gene set enrichment analysis and immune cell infiltration assessment supported the model's efficacy. The results from both the univariate and multivariate analyses suggest that the risk score is an independent prognostic factor with a hazard ratio of 2.97 (95% CI, 2.28 to 3.9, P < .001). In the validation cohort, the AUC at 1, 2, and 3 years is 0.74, 0.74, and 0.72, respectively. Our findings proposed biomarkers with prognostic potential, laying the groundwork for future in vitro validation and therapeutic exploration. This contributes to a deeper understanding of the genes associated with bladder TME and may improve prognostic precision in BLCA management.

ROLE OF HAEMATOLOGY BIOMARKERS IN URINARY BLADDER AND RENAL CELL CANCER PATIENTS TREATED WITH IMMUNE-ONCOLOGICAL DRUGS

Background: Urinary bladder (UB) and Renal Cell Cancer (RCC) are common in men than women with poor outcome. The novel immunotherapy drugs like Immune Check Point Inhibitors (ICIs) are effective but expensive. However, a cost-effective and reliable biomarker to predict response and clinical outcomes is lacking. Objective: To study the association of pre-treatment haematological parameters with clinical outcome in urinary bladder and renal cell cancer patients treated with ICIs. Method: In a retrospective study, we included 52 patients with urinary bladder and RCC treated with ICIs from Jan 2008 to Dec 2019. Clinical evaluation and laboratory investigations were performed as a part of standard protocol. CBC parameters such as WBC, TLC, DLC, Hb, Platelet, Neutrophil:Lymplocyte Ratio(NLR), Platelet:Lymphocyte Ratio (PLR), Lymphocyte:Monocyte Ratio(LMR) were studied at the time of TMC enrolment(first) and before the start (pre-treatment) of the ICIs therapy(io). Results: Amongst the CBC parameters, WBC count, when categorized based on cut off from ROC at the time of TMC enrolment(first) and the pre-treatment of the ICIs therapy(io) were found to be significantly associated with progression of disease with p- value 0.012 with Hazard Ratio (HR) - 2.52 (95% C.I. - 1.2-5.31) &amp; p- value 0.060 with Hazard Ratio (HR) – 1.99 (95% C.I. – 0.96-4.12) respectively. The ROC - based cut off for WBC at the time of TMC enrolment(first) and the pre-treatment of the ICIs therapy(io) were found to be 7.16 X10e3/µL (AUC of 72%) with 71% sensitivity and 72% specificity and 6.48 X10e3/µL (AUC of 74%) with 67% sensitivity and 72% specificity respectively to predict progressed cases with respect to non-progressed cases. Along with WBC, the pre-treatment Absolute Neutrophil Count (ANC) (&gt;3.67)&amp; Absolute Monocyte count (AMC)(&gt;0.42) pre-treatment of the ICIs therapy(io)) showed a significance with Hazard Ratio (HR) – 3.09(95% C.I.– 1.26-7.6) with p-value 0.010, Hazard Ratio (HR) – 3.48(95% C.I. - 1.6-7.57)and p-value &lt;0.001 respectively. The ROC- based cut off for ANC at the pre-treatment of the ICIs therapy(io) were found to be 3.67 X10e3/µL (AUC of 68.4%) with 82.4% sensitivity and 50% specificity. Similarly The ROC- based cut off for AEC at the time of TMC enrolment(first) and AMC at the pre- treatment of the ICIs therapy(io) were found to be 0.22 X10e3/µL (AUC of 69.6%) with 55.9% sensitivity and 82.3% specificity and 0.42 X10e3/µL (AUC of 74.7%) with 72.7% sensitivity and 72.2% specificity respectively to predict progressed cases compared to non-progressed cases. Of 52 patients, 34 (65.4%) had progression. The median (m) PFS was 8.67 months (95% CI 2.26 - 15.09).The one year PFS rate was 46.5 (95% CI 31.2 -60.5). The mPFS in WBC &lt;=7.16 was 26.48 months (95% CI 0 - 53.08) higher than the mPFS of 5.75 (95% CI 0.21 - 11.29) in WBC &gt;7.16 with P value 0.012 at the time of TMC enrolment. Similarly, the mPFS in WBC pre- treatment of the ICIs therapy(io))&lt;=6.48 was13.11 months (95% CI 0 – 28.95) higher than the mPFS of 6.18 (95% CI 2.67-9.68) in WBC &gt;6.48 with P value 0.060.Along with WBC, the mPFS in ANC at the pre- treatment of the ICIs therapy(io)&lt;=3.67 was 27.4(95% CI 0-55.21) higher than the mPFS of 6.18(95% CI 1.47-10.88)in ANC &gt;3.67 with p value 0.010. The mPFS in AMC at the pre-treatment of the ICIs therapy(io)&lt; =0.415 was 27.4 (95% CI 9.41-45.39) higher than the mPFS of 3.91 (95% CI 0.06-7.76) in AMC &gt;0.415with p- value &lt;0.001.We observed mPFS in immunotherapy cycles &gt;=6 to be 15.9 (95% CI 6.98-24.82) was higher than the mPFS of 2.23 (95% CI 0.99-3.481) in immunotherapy cycles&lt;6 with p value 0.025. The patients were followed for a period of 40.35 months (95% CI 26.98 - 53.71). Of 52 patients, 32(61.5%) patients deceased with median OS of 18.46 months (95% CI 10.12 - 26.80) and the one year OS rate was 57.6 (95% CI 41.6 - 70.6). Conclusion: Simple, routinely available and cost effective WBC (TLC), Absolute Neutrophil Count (ANC) ,Absolute Monocyte Count (AMC) and Absolute Eosinophil Count (AEC) from CBC test has a great potential to be used as a biomarker to predict response and clinical outcomes in patients with urinary bladder cancer and RCC patients treated with ICIs.

Polo-like kinase 1 inhibition modulates urinary tract smooth muscle contraction and bladder cell transcriptional programs.

The serine/threonine kinase polo-like kinase 1 (PLK1) is a master regulator of cell proliferation and contraction, but its physiological role in the lower urinary tract is unknown. We utilized transcriptomic programs of human bladder smooth muscle cells (hBSMCs), 3D bladder spheroid viability assays, and human ureterovesical junction contractility measurements to elucidate the impacts of PLK1 inhibition. This work reveals PLK1 reduction with the selective inhibitor TAK-960 (500 nM) suppresses high K+-evoked contractions of human urinary smooth muscle ex vivo while decreasing urothelial cell viability. Transcriptomic analysis of hBSMCs treated with TAK-960 shows modulation of cell cycle and contraction pathways, specifically through altered expression of Cys2/His2-type zinc finger transcription factors. In bladder spheroids, PLK1 inhibition also suppresses smooth muscle contraction protein filamin. Taken together, these findings establish PLK1 is a critical governor of urinary smooth muscle contraction and urothelial proliferation with implications for lower urinary tract disorders. Targeting PLK1 pharmacologically may therefore offer therapeutic potential to ameliorate hypercontractility and aberrant growth. Further elucidation of PLK1 signaling networks promises new insights into pathogenesis and much needed treatment advances for debilitating urinary symptoms.

A New Strategy to Authenticate Prognosis and Tumor Immunity Model Based on Cancer-Associated Fibroblasts lncRNA in Bladder Cancer

There is growing evidence indicating that long noncoding RNAs (lncRNAs) play a pivotal role in regulating both cancer immunity and the cancer microenvironment. Specifically, cancer-associated fibroblasts (CAFs) have been found to have a significant impact on a variety of tumor types. However, the specific lncRNAs associated with CAFs have not yet been thoroughly studied in the context of bladder cancer. This study examined specific lncRNAs derived from TCGA data for the bladder tumor immune microenvironment (TME). Through our analysis, we identified 67 fibroblast-specific lncRNAs (referred to as FIBLnc), and further identified ten gene signatures that include RGS5, EPB41L4A-AS1, SNHG5, MAGI2-AS3, PVT1, EMX2OS, NEAT1, DLEU1, TMEM99, and A1BG-AS1. To establish the robustness of our findings, we also analyzed the gene expression profiles in the GSE13507, GSE19915, GSE31684, GSE48276, and GSE48277A datasets. The results from these datasets, along with the TCGA dataset, were highly accurate in detecting differences in overall survival using the FIBLnc signatures. In functional analysis, FIBLnc revealed its association with immune checkpoints and tumor progression. Furthermore, patients who were treated with anti-PD-1 or anti-CTLA4 could benefit from the FIBLnc score to predict their immunotherapy response. We further investigated the expression and prognostic power of lncRNAs EPB41L4A-AS1, TMEM99, and A1BG-AS1 in TCGA, bladder cell lines, and 24 paired tissues. Our results showed lower expression of EPB41L4A-AS1 and higher expression of TMEM99 and A1BG-AS1 in bladder malignancy, which correlated with a worse prognosis. Furthermore, we found that lncRNAEPB41L4A-AS1 could inhibit bladder cell carcinoma invasion in vitro. Overall, our study highlights the value of FIBLnc in assessing the immune environment status of individuals with bladder cancer and predicting their immunotherapy response. This investigation sheds light on the intricate processes involved in bladder cancer and underscores the importance of considering the FIBLnc score in the prognosis and management of this malignant tumor.

Programmed Cell Death Ligand 1 (PD-L1) Immunohistochemical Expression in Advanced Urothelial Bladder Carcinoma: An Updated Review with Clinical and Pathological Implications.

The management of advanced bladder carcinoma involves a multidisciplinary approach, but the prognosis remains poor for many patients. The immune system plays a crucial role in this disease, influencing both tumor development and response to treatment, and exploiting the immune system against the tumor can be a valuable strategy to destroy neoplastic cells. This is the biological principle underlying Bacillus Calmette-Guérin (BCG) use and, more recently, immune checkpoint inhibitors (ICIs), like PD-1 (programmed death-1)/PD-L1 (programmed death-ligand 1) inhibitors. In fact, one of the best studied immune checkpoints is represented by the PD-1/PD-L1 axis, which is a well-known immune escape system adopted by neoplastic bladder cells. PD-L1 expression has been associated with a higher pathologic stage and has shown prognostic value in bladder carcinoma. Interestingly, high-grade bladder cancers tend to express higher levels of PD-1 and PD-L1, suggesting a potential role of such an axis in mediating disease progression. Immunotherapy with PD-1 and PD-L1 inhibitors has therefore emerged as a valuable treatment option and has shown efficacy in advanced bladder cancer patients, with high PD-L1 expression levels associated with better treatment responses. Our review aims to provide a comprehensive overview of the role of PD-L1 in advanced bladder cancer, focusing on its implications for treatment decisions and the prediction of treatment response. Overall, our work aims to contribute to the understanding of PD-L1 as a predictive biomarker and highlight its role in shaping therapeutic approaches for advanced bladder cancer.

Census tract–level social determinants of health and variability in stage at diagnosis of bladder, breast, and non-small cell lung cancers.

11170 Background: Social determinants of health (SDOH) can identify underserved populations, inform the patient journey, and identify opportunities to improve outcomes. In cancer, SDOH may influence stage at diagnosis through exposure to risk factors for aggressive cancer and decreased access to care. However, privacy restrictions can limit research with SDOH in real world datasets, which often only know a patient’s broad location. Methods: Geocoding was undertaken on addresses of patients diagnosed between 1/1/17 and 9/30/22 with bladder (n=6078), breast (n=28,958), and non-small cell lung cancer (NSCLC, n=14,957) in the Syapse Learning Health Network of U.S. community health systems. Location was linked to five indicators of SDOH at the census tract level: the Social Vulnerability Index (SVI); percent (%) of tract spending &gt;30% of income on housing; % of tract with broadband internet; county designation as a primary care shortage area; and rural urban commuting area (RUCA). Nested multivariable ordinal logistic regression models estimated independent associations between demographic, clinical, and SDOH factors with stage at initial cancer diagnosis. The statistical significance of including SDOH variables was assessed using a likelihood ratio test (LRT) comparing the models before and after including SDOH. Results: After successful geocoding and linkage to census tracts, there were marginal differences in the distribution of the five SDOH across stages at diagnosis within the three tumor types. In multivariable models adjusted for year of diagnosis, age, sex, race, ethnicity, smoking, and primary payor, among the tested SDOH measures, county designated as primary care shortage (proportional odds ratios (OR): 1.31, 95% confidence interval (95% CI): 1.05-1.64), and non-metropolitan area (0.69, 0.50-0.94) were statistically significantly associated with stage at bladder cancer diagnosis. SVI (1.52, 1.31-1.76) was also statistically significantly associated and directly proportional with stage at breast cancer diagnosis. Among patients with NSCLC, broadband internet access (1.01, 1.00-1.01), primary care shortage (1.15, 1.03-1.27) and non-metropolitan area (0.85, 0.74-0.99) were statistically associated with stage at diagnosis. In all three tumors, LRTs found that models with SDOH were statistically significantly better at predicting stage at diagnosis than those without (p &lt;0.001). Conclusions: Using granular, census-tract geospatial resolution for geocoding of SDOH, we find that patients who live in greater vulnerability are diagnosed at later stages than those who live in areas of less vulnerability, suggesting that consideration of SDOH variables is important in assessing stage at diagnosis and identify patients with an unmet need for outreach and screening.

Impact of carboplatin and cisplatin shortages on treatment patterns in patients with metastatic solid tumors.

11149 Background: The United States experienced cisplatin (CP) and carboplatin (CB) shortages in 2023, leading to potential rationing or switching to alternative therapies in multiple cancers. Using a US nationwide, real-world oncology dataset, we assessed changes in the use of CP and CB in 7 cancers during the shortage period. Methods: All patients (pts) from the Flatiron Health electronic health record-derived de-identified database (~280 cancer clinics, ~800 sites of care) with a machine learning-extracted metastatic (met) diagnosis of a cancer of interest between March 2022 and June 2023 and evidence of first-line (1L) therapy, were selected. Pts with multiple primaries were excluded. Platinum of interest was identified based on frequency of pre-period 1L use for a given cancer. The pre-period was June 2022 through first month of reported shortage (February 2023 for CP; April 2023 for CB); data cutoff was June 2023. Clinical characteristics of platinum-treated pts stratified by pre- vs post-period by disease were assessed. 1L platinum utilization rates (UR), calculated as the proportion of 1L-treated pts initiating CB/CP within 30 days of met diagnosis, were plotted by month. Cohort-level impact was evaluated with an interrupted time series analysis—overall and stratified by practice type, practice size, and socioeconomic status (SES). Multiple testing was controlled using the Benjamini-Hochberg procedure. Results: 10,983 pts received 1L therapy in the study period. Monthly platinum UR were stable across diseases in the pre-period. However, there was a significant post-period decrease in CB usage for non-small cell lung (NSCLC) and endometrial cancers (EC), and a decreasing trend in UR for CB in bladder (BC), ovarian (OC), and small cell lung cancers (SCLC). We also observed a decreasing trend in CP usage for cholangiocarcinoma (CC). Post-period monthly UR were stable for CB and CP in head and neck cancer (HNC), and for CP in BC. Odds ratios (OR) representing the month-over-month change in the odds of receiving platinum therapy during both pre- and post-periods are shown (table). OR trends varied by practice type and disease; no notable trends were seen by practice size or SES. UR with additional follow-up times will be presented. Conclusions: We observed decreased platinum use across multiple cancers during the shortage period, most notably for met NSCLC and EC, suggesting shortages led oncologists to seek alternatives to standard 1L platinum-based regimens. Further study of how treatment changes due to platinum shortages impacted pt outcomes is warranted. [Table: see text]

A phase I/II, first-in-human study of VLS-1488, an oral KIF18A inhibitor, in patients with advanced cancer.

TPS3181 Background: Chromosomal instability (CIN) is a hallmark of cancer characterized by high rates of chromosomal segregation errors during mitosis. This creates structural and numerical changes to chromosomes, driving genetic heterogeneity and cancer evolution. KIF18A is a mitotic kinesin protein shown to be important for successful division of cancer cells with CIN but not required for mitosis in normal diploid cells. Pre-clinical studies demonstrated that treatment of chromosomally unstable cancer cells in vitro and in xenograft models (HCC15 and OVCAR-3) with a KIF18A inhibitor resulted in failure of chromosomal congression, mitotic arrest, cell death and dose-dependent inhibition of tumor growth. VLS-1488 is an oral small molecule inhibitor of KIF18A that is being examined in an open-label, phase I/II, first-in-human study evaluating the safety, tolerability, pharmacokinetics (PK) and preliminary anti-tumor activity in advanced solid tumors with high prevalence of CIN. Methods: Key eligibility criteria include specific histologically confirmed metastatic or advanced tumor types with at least one site of measurable disease per RECIST 1.1, adequate organ function, and mismatch repair proficient/microsatellite stable.Eligible tumor types include high grade serous ovarian/fallopian tube/primary peritoneal cancer, squamous non-small cell lung cancer, triple negative breast cancer, gastric adenocarcinoma, gastroesophageal cancer, esophageal cancer, colorectal adenocarcinoma, transitional cell carcinoma of bladder, head and neck squamous cell carcinoma, ovarian or uterine carcinosarcoma, and uterine serous carcinoma. Subjects will receive VLS-1488 monotherapy continuously for 28-day cycles once daily. The study consists of two parts, Dose Escalation and Dose Expansion. Utilizing a Bayesian Optimal Interval (BOIN) design, Dose Escalation will examine the safety and tolerability of VLS-1488 at various dose levels (DLs) to identify the Maximum Tolerated Dose (MTD) and to select DLs for Dose Expansion. Dose Expansion may commence after the MTD or DLs of interest are identified. Dose Expansion will examine the safety, tolerability, preliminary efficacy and preliminary drug-drug interaction (DDI) and food effect risk of VLS-1488 in selected tumor types. Subjects enrolled to specific tumor type cohorts may be randomized to DLs of interest. Enrollment to Dose Escalation began in September 2023 and is ongoing. Clinical trial information: NCT05902988 .

Evaluating birch leaf tea as a functional herbal beverage: Beneficial impact on the urinary tract, and metabolism in human organism

Herbal teas are considered as a potential constituent of novel functional beverages consumed daily. One of the commonly used herbal teas is silver birch (Betula pendula Roth) leaf infusion, traditionally used in urinary tract diseases. In this study, the potential of birch leaf infusion as a functional beverage, emphasizing its active ingredients’ bioavailability, anti-inflammatory, and antiadhesive properties concerning urinary tract health, was investigated. A complex approach was proposed, which included phytochemical screening, bioavailability, gut microbiota biotransformation, and an in vivo test for urine metabolomics assessment. The bioassays confirmed significant anti-inflammatory (interleukins IL-6 and IL-8 secretion) and anti-adhesive (Uropathogenic Escherichia coli and T24 bladder cells) activities. The high-resolution mass spectrometry metabolomics studies linked gut microbiota metabolites and the metabolites present in the urine. Several metabolites connected with phenolics’ consumption were detected in the urine, e.g., glucuronides and sulfates of caffeic acid and dihydroxyphenyl-γ-valerolactones. Based on the presented results, the birch leaf should be considered useful in designing functional beverages, especially targeted to the groups at high risk of urinary diseases.

Abstract A016: NRF2 activation promotes a fitness disadvantage in normal urothelium and drives a basal-like phenotype

Abstract Introduction: Nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor that plays a key role in protecting cells from oxidative stress. NRF2 is negatively controlled by Kelch-like ECH-associated protein 1 (KEAP1), which facilitates NRF2 ubiquitination and proteosomal degradation under homeostatic conditions. NRF2 activating mutations occur at a frequency of 5.9% in muscle-invasive bladder cancer (MIBC). However, the role of NRF2 in MIBC has not been fully understood. The aim of this study is to clarify how NRF2 affect to the normal epithelial cells and the tumor cells in bladder. Methods: We used genetically engineered mice (GEM) with a Cre inducible Nrf2 activating mutation (LSL-Nrf2-E79Q). To evaluate how Nrf2 affects the tumorigenesis process in vivo, the N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN) was administered to Upk3a-CreERT2; LSL-Nrf2-E79Q/+ (UN) mice. We also established normal urothelial organoids and BBN tumor derived organoids from wild-type (WT) and Krt5-CreERT2; LSL-Nrf2-E79Q/+ (K5N) mice for ex vivo study. To induce overexpression of NRF2, we utilized KI696 (KEAP1-NRF2 inhibitor) as a pharmacological agent and performed NAD/NADH assay to quantify the reductive stress. Additionally, we performed single cell RNA sequencing (scRNA-seq) with WT, UN and K5N mice bladder to identify the effect of Nrf2E79Q on the normal epithelial cells. Results: In BBN induced tumor model in vivo, UN mice did not differ in time to tumor development from the control WT mice. Furthermore, UN tumors had lost the post-recombination allele of LSL-Nrf2-E79Q in genomic PCR, meaning cells expressing Nrf2E79Q are selected against during the process. In ex vivo study, KI696 induced reductive stress in both WT normal urothelial organoids and BBN tumor derived organoids, but inhibited cell proliferation only in normal urothelial organoids. In K5N organoids, Nrf2E79Q activation enhanced relative mRNA expression of a number of basal related genes. In addition to that, vehicle organoids in differentiation media showed phenotypic differentiation, whereas those treated with tamoxifen did not demonstrate phenotypic signs of differentiation suggesting Nrf2 activation promotes a basal-like phenotype. Furthermore, we annotated three clusters in mice bladder epithelial cells as luminal, intermediate and basal using scRNA-seq and looked at their proportion among WT, UN and K5N mice. We found that the percentage of luminal cells was decreased and intermediate and basal cells increased in both UN and K5N mice compared to WT mice. This indicated that Nrf2E79Q promotes disadvantage in luminal cells in UN mice, whereas it contributes to the increase in intermediate and basal cells in K5N mice. Conclusion: These results suggested that Nrf2E79Q promotes a fitness disadvantage in normal mice urothelium. Furthermore, Nrf2E79Q inhibits differentiation of K5N organoids and drives a basal-like phenotype in GEM models. Further investigation is needed to clarify the mechanism of NRF2 activating mutation contributing the tumorigenesis in bladder. Citation Format: Akihiro Hamada, Yuki Kita, Di Wu, Mi Zhou, Sean Clark-Garvey, Andrew Gdowski, Michael Sturdivant, Wolfgang Beckabir, Elliott Drew Toomer, Emily Kounlavong, Lucia Lim, Kathryn Gessner, Jeffrey Damrauer, William Kim. NRF2 activation promotes a fitness disadvantage in normal urothelium and drives a basal-like phenotype [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr A016.

Circular RNA LMBR1 inhibits bladder cancer progression by enhancing expression of the protein ALDH1A3

BackgroundCircular RNAs (circRNAs) have been identified as playing an integral role in the development of bladder cancer (BC). However, the mechanism by which circRNAs operate in the chemical carcinogenesis of BC remains unclear. MethodsTo explore this mechanism, we used RNA high-throughput sequencing to identify differentially expressed circRNA in bladder epithelial cells and chemically induced malignant transformed BC cells. Subsequently, in vitro experiments were conducted to investigate the biological function and molecular mechanism of circLMBR1 in BC. Finally, animal experiments were conducted to examine the clinical relevance of circLMBR1 in vivo. ResultsOur profiling of circular RNA expression during cellular malignant transformation induced by chemical carcinogens identified a subset of circRNAs associated with cell transformation. We verified that the expression of circLMBR1 in bladder epithelial malignant transformed cells was decreased compared with control cells, as well as in BC tissues and bladder cell lines. Furthermore, circLMBR1 was seen to inhibit the proliferation, invasion, and migration of BC cells both in vitro and in vivo. Mechanistically, circLMBR1 was found to exert its antitumor effect by binding to the protein ALDH1A3. ConclusionsOur findings have revealed that circLMBR1 inhibits the progression of BC cells by binding to ALDH1A3 and upregulating its expression. As such, circLMBR1 serves as a promising predictor of BC and may provide a novel therapeutic target for the treatment of BC.

Gentamicin loaded niosomes against intracellular uropathogenic Escherichia coli strains

Urinary tract infections (UTIs) are the most common bacterial infections and uropathogenic Escherichia coli (UPEC) is the main etiological agent of UTIs. UPEC can persist in bladder cells protected by immunological defenses and antibiotics and intracellular behavior leads to difficulty in eradicating the infection. The aim of this paper is to design, prepare and characterize surfactant-based nanocarriers (niosomes) able to entrap antimicrobial drug and potentially to delivery and release antibiotics into UPEC-infected cells. In order to validate the proposed drug delivery system, gentamicin, was chosen as “active model drug” due to its poor cellular penetration. The niosomes physical–chemical characterization was performed combining different techniques: Dynamic Light Scattering Fluorescence Spectroscopy, Transmission Electron Microscopy. Empty and loaded niosomes were characterized in terms of size, ζ-potential, bilayer features and stability. Moreover, Gentamicin entrapped amount was evaluated, and the release study was also carried out. In addition, the effect of empty and loaded niosomes was studied on the invasion ability of UPEC strains in T24 bladder cell monolayers by Gentamicin Protection Assay and Confocal Microscopy. The observed decrease in UPEC invasion rate leads us to hypothesize a release of antibiotic from niosomes inside the cells. The optimization of the proposed drug delivery system could represent a promising strategy to significatively enhance the internalization of antimicrobial drugs.