Surface Labeling with Adhesion Protein FimH Improves Binding of Immunotherapeutic Agent Salmonella Ty21a to the Bladder Epithelium

2020 ◽  
pp. 1-12
Author(s):  
Maroeska J. Burggraaf ◽  
Lisette Waanders ◽  
Mariska Verlaan ◽  
Janneke Maaskant ◽  
Diane Houben ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Antitumor Activity ◽  
Bladder Wall ◽  
Bladder Epithelium ◽  
Modest Improvement ◽  
Adhesion Protein ◽  
Survival Experiment ◽  
Muscle Invasive

BACKGROUND: Bladder cancer is the ninth most common cancer in men. 70% of these tumors are classified as non-muscle invasive bladder cancer and those patients receive 6 intravesical instillations with Mycobacterium bovis BCG after transurethral resection. However, 30% of patients show recurrences after treatment and experience severe side effects that often lead to therapy discontinuation. Recently, another vaccine strain, Salmonella enterica typhi Ty21a, demonstrated promising antitumor activity in vivo. Here we focus on increasing bacterial retention in the bladder in order to reduce the number of instillations required and improve antitumor activity. OBJECTIVE: To increase the binding of Ty21a to the bladder wall by surface labeling of the bacteria with adhesion protein FimH and to study its effect in a bladder cancer mouse model. METHODS: Binding of Ty21a with surface-labeled FimH to the bladder wall was analyzed in vitro and in vivo. The antitumor effect of a single instillation of Ty21a+FimH in treatment was determined in a survival experiment. RESULTS: FimH-labeled Ty21a showed significant (p <  0.0001) improved binding to mouse and human cell lines in vitro. Furthermore, FimH labeled bacteria showed ∼5x more binding to the bladder than controls in vivo. Enhanced binding to the bladder via FimH labeling induced a modest improvement in median but not in overall mice survival. CONCLUSIONS: FimH labeling of Ty21a significantly improved binding to bladder tumor cells in vitro and the bladder wall in vivo. The improved binding leads to a modest increase in median survival in a single bladder cancer mouse study.

Pembrolizumab and chemoradiotherapy for muscle invasive bladder cancer: The ANZUP 1502 PCR-MIB trial.

2018 ◽  
Vol 36 (6_suppl) ◽  
pp. TPS531-TPS531 ◽  
Author(s):  
Andrew James Weickhardt ◽  
Farshad Foroudi ◽  
Shomik Sengupta ◽  
Laura Galletta ◽  
Alan Herschtal ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Molecular Genetic ◽  
Response To Therapy ◽  
Invasive Bladder Cancer ◽  
Muscle Invasive Bladder Cancer ◽  
Bladder Preservation ◽  
Immunological Parameters ◽  
Muscle Invasive

TPS531 Background: Pembrolizumab leads to responses in ~20% of metastatic bladder cancer patients. Irradiation of bladder cancer cells in-vitro and in-vivo leads to upregulation of PD-L1, and in immunocompetent mouse models blockade of PD-L1 leads to delayed tumour growth following irradiation. Randomised data from PACIFIC trial in NSCLC shows the addition of PD-L1 inhibition to chemoradiation significantly prolongs PFS. A trial of chemoradiotherapy with pembrolizumab will assess safety and synergy of the combination in localised bladder cancer. Methods: This pilot study enrols patients with maximally resected non-metastatic muscle invasive bladder cancer, who either wish for bladder preservation or are ineligible for cystectomy. This study will assess the safety and feasibility of combining pembrolizumab with chemoradiotherapy in ECOG 0-1 patients without contraindications to pembrolizumab. The study has enrolled 4 of a planned 30 patients. All patients treated with 64Gy of radiation therapy in 32 fractions over 6 weeks, 2 days. Cisplatin 35mg/m2 IV concurrently weekly for 6 doses with radiation. Pembrolizumab commences concurrently with radiation and is given 200mg IV q21 days for 7 doses. Surveillance cystoscopy is performed 12 & 24 weeks after the commencement of chemoradiotherapy to assess response to therapy. Patients will enter follow up with clinical assessment, cystoscopy and CT staging performed at intervals until close of study. The primary endpoint assessed will be safety, as defined by a satisfactorily low rate of unacceptable toxicity (G3-4 adverse events or failure of completion of planned chemotherapy and radiotherapy according to defined parameters). The secondary endpoint will be efficacy, as assessed by the proportion of patients achieving a best response of complete response based on the first two 12 and 24 week post chemoradiotherapy cystoscopic assessments. Exploratory analysis will include assessment of tumour histopathological, molecular, genetic and immunological parameters. It is expected that it will take two years to accrue the 30 patients across 5 Australian centres. NCT02662062. Clinical trial information: NCT02662062.

scholarly journals Nanaomycin K inhibited epithelial mesenchymal transition and tumor growth in bladder cancer cells in vitro and in vivo

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Koichi Kitagawa ◽  
Katsumi Shigemura ◽  
Aya Ishii ◽  
Takuji Nakashima ◽  
Hirotaka Matsuo ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cancer Cells ◽  
Epithelial Mesenchymal Transition ◽  
Mesenchymal Transition ◽  
Invasion And Migration ◽  
Bladder Cancer Cells ◽  
And Migration ◽  
Muscle Invasive

AbstractNanaomycin K, derived from Streptomyces rosa subsp. notoensis OS-3966T, has been discovered to have inhibitory bioactivity on epithelial–mesenchymal transition (EMT), an important mechanism of cancer cell invasion and migration. In this study, we examined the anti-EMT and anti-tumor effect of nanaomycin K in bladder cancer, where EMT has important roles in progression. We treated two bladder cancer lines, non-muscle-invasive KK47 and muscle-invasive T24, with nanaomycin K to determine the effects on cell proliferation, apoptosis and expression of EMT markers in vitro. Wound-healing assays were performed to assess cell invasion and migration. We conducted an in vivo xenograft study in which mice were inoculated with bladder cancer cells and treated with intratumoral administration of nanaomycin K to investigate its anti-tumor and EMT inhibition effects. As the results, nanaomycin K (50 µg/mL) significantly inhibited cell proliferation in KK47 (p < 0.01) and T24 (p < 0.01) in the presence of TGF-β, which is an EMT-inducer. Nanaomycin K (50 µg/mL) also significantly inhibited cell migration in KK47 (p < 0.01) and T24 (p < 0.01), and induced apoptosis in both cell lines in the presence of TGF-β (p < 0.01). Nanaomycin K increased the expression of E-cadherin and inhibited the expression of N-cadherin and vimentin in both cell lines. Nanaomycin K also decreased expression of Snail, Slug, phospho-p38 and phospho-SAPK/JNK especially in T24. Intratumoral administration of nanaomycin K significantly inhibited tumor growth in both KK47 and T24 cells at high dose (1.0 mg/body) (p = 0.009 and p = 0.003, respectively) with no obvious adverse events. In addition, nanaomycin K reversed EMT and significantly inhibited the expression of Ki-67 especially in T24. In conclusion, we demonstrated that nanaomycin K had significant anti-EMT and anti-tumor effects in bladder cancer cells, suggesting that nanaomycin K may be a therapeutic candidate for bladder cancer treatment.

Biogenic silver nanoparticles: In vitro and in vivo antitumor activity in bladder cancer

2020 ◽  
Vol 151 ◽  
pp. 162-170 ◽  
Author(s):  
Luiz Alberto Bandeira Ferreira ◽  
Fernanda Garcia-Fossa ◽  
Allan Radaic ◽  
Nelson Durán ◽  
Wagner José Fávaro ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Silver Nanoparticles ◽  
Antitumor Activity ◽  
Biogenic Silver Nanoparticles ◽  
In Vivo Antitumor Activity

scholarly journals The Antitumor Effects of Plasma-Activated Saline on Muscle-Invasive Bladder Cancer Cells In Vitro and In Vivo Demonstrate Its Feasibility as a Potential Therapeutic Approach

Cancers ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1042
Author(s):  
Hao Zhang ◽  
Jishen Zhang ◽  
Bo Guo ◽  
Hailan Chen ◽  
Dehui Xu ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cancer Cells ◽  
Culture Media ◽  
Invasive Bladder Cancer ◽  
Antitumor Effects ◽  
Muscle Invasive Bladder Cancer ◽  
Plasma Irradiation ◽  
Muscle Invasive

Muscle-invasive bladder cancer (MIBC) is a fast-growing and aggressive malignant tumor in urinary system. Since chemotherapy and immunotherapy are only useable with a few MIBC patients, the clinical treatment of MIBC still faces challenges. Here, we examined the feasibility of plasma-activated saline (PAS) as a fledgling therapeutic strategy for MIBC treatment. Our data showed that plasma irradiation could generate a variety of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in saline. In vivo tests revealed that pericarcinomatous tissue injection with PAS was effective at preventing subcutaneous bladder tumor growth, with no side effects to the visceral organs after long-term administration, as well as having no obvious influence on the various biochemistry indices of the blood in mice. The in vitro studies indicated that adding 30% PAS in cell culture media causes oxidative damage to the bladder transitional cells T24 and J82 through enhancing the intracellular ROS level, and eventually induces cancer cells’ apoptosis by activating the ROS-mediated Fas/CD95 pathway. Therefore, for an intracavity tumor, these initial observations suggest that the soaking of the tumor tissue with PAS by intravesical perfusion may be a novel treatment option for bladder cancer.

scholarly journals Combining mTOR Inhibition with Radiation Improves Antitumor Activity in Bladder Cancer Cells In Vitro and In Vivo: A Novel Strategy for Treatment

PLoS ONE ◽  
2013 ◽  
Vol 8 (6) ◽  
pp. e65257 ◽  
Author(s):  
Roland Nassim ◽  
Jose Joao Mansure ◽  
Simone Chevalier ◽  
Fabio Cury ◽  
Wassim Kassouf
Keyword(s):  
Bladder Cancer ◽  
Antitumor Activity ◽  
Cancer Cells ◽  
Mtor Inhibition ◽  
Bladder Cancer Cells ◽  
Novel Strategy

Evaluation of a Novel Cystoscopic Compatible Cryocatheter for the Treatment of Bladder Cancer

2020 ◽  
Vol 6 (3) ◽  
pp. 303-318
Author(s):  
John M. Baust ◽  
Anthony Robilotto ◽  
Kimberly L. Santucci ◽  
Kristi K. Snyder ◽  
Robert G. Van Buskirk ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Bladder Wall ◽  
Thermal Characteristics ◽  
Fluorescent Imaging ◽  
Lesion Depth ◽  
3 Dimensional ◽  
Study Results ◽  
Freeze Time

BACKGROUND: As the acceptance of cryoablative therapies for the treatment of non-metastatic cancers continues to grow, avenues for novel cryosurgical technologies and approaches have opened. Within the field of genitourinary tumors, cryosurgical treatments of bladder cancers remain largely investigational. Current modalities employ percutaneous needles or transurethral cryoballoons or sprays, and while results have been promising, each technology is limited to specific types and stages of cancers. OBJECTIVE: This study evaluated a new, self-contained transurethral cryocatheter, FrostBite-BC, for its potential to treat bladder cancer. METHODS: Thermal characteristics and ablative capacity were assessed using calorimetry, isothermal analyses, in vitro 3-dimensional tissue engineered models (TEMs), and a pilot in vivo porcine study. RESULTS: Isotherm assessment revealed surface temperatures below – 20°C within 9 sec. In vitro TEMs studies demonstrated attainment of ≤– 20°C at 6.1 mm and 8.2 mm in diameter following single and double 2 min freezes, respectively. Fluorescent imaging 24 hr post-thaw revealed uniform, ablative volumes of 326.2 mm3 and 397.9 mm3 following a single or double 2 min freeze. In vivo results demonstrated the consistent generation of ablative areas. Lesion depth was found to correlate with freeze time wherein 15 sec freezes resulted in ablation confined to the sub-mucosa and ≥30 sec full thickness ablation of the bladder wall. CONCLUSIONS: These studies demonstrate the potential of the FrostBite-BC cryocatheter as a treatment option for bladder cancer. Although preliminary, the outcomes of these studies were encouraging, and support the continued investigation into the potential of the FrostBite-BC cryocatheter as a next generation, minimally invasive cryoablative technology.

Augmentation of antitumor activity of 5′-deoxy-5-fluorouridine by thymosin fraction 5 in mouse bladder cancer cells in vitro and in vivo

1999 ◽  
Vol 145 (1-2) ◽  
pp. 121-126 ◽  
Author(s):  
Shinichi Ikemoto ◽  
Kazunobu Sugimura ◽  
Seiji Wada ◽  
Ryouji Yasumoto ◽  
Keisuke Yamamoto ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Antitumor Activity ◽  
Cancer Cells ◽  
Bladder Cancer Cells

scholarly journals The Interplay Between Oxidative Phosphorylation and Glycolysis as a Potential Marker of Bladder Cancer Progression in Vitro and in Vivo

2020 ◽  
Author(s):  
Greta Petrella ◽  
Giorgia Ciufolini ◽  
Giusy Burgio ◽  
Andrea Salonia ◽  
Francesco Montorsi ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
High Risk ◽  
Oxidative Phosphorylation ◽  
Cell Lines ◽  
Low Grade ◽  
High Grade ◽  
Risk Of Progression ◽  
Muscle Invasive

Abstract BackgroundUrothelial bladder cancer (UBC) is the most common tumor of the urinary system, the ninth most common cancer worldwide and the one with the most expensive treatment from diagnosis to death. One of the biggest problems related to this disease is the lack of sufficiently accurate markers that can anticipate the progression of the cancer from a low-grade non-muscle invasive to a high-grade muscle invasive UBC. Genomics and transcriptomics have recently added a number of molecular markers to traditional observations based on pathological parameters, which have greatly improved the prediction of risk of recurrence and progression. The inclusion of information from other omics sciences, such as metabolomics, could significantly improve this scenario.MethodsIn this study, we present the metabolic characterization using 1H-NMR of three UBC cell lines representing tumors with low-risk of progression, RT4, high-risk, 5637, and a cell line that shares characteristics with both, RT112. The metabolic profiles were classified by multivariate analysis. To validate the in vitro results, concentrations of two metabolites were measured in vivo in the urine of 91 patients with non-invasive and invasive tumors.ResultsRT4 cells mainly use oxidative phosphorylation to produce ATP and biomass, 5637 cells depend mainly on glycolysis, while RT112 cells show a mixed state with both metabolisms partially activated. The lactate/alanine ratio proved to be the most sensitive marker to the different type of metabolism active in the cells in vitro. By measuring its value in vivo in urine, we have found a two-fold increase among patients with high-grade tumors compared to low-grade ones.ConclusionsOur results reveal for the first time the relative importance of glycolysis and oxidative phosphorylation in the growth and maintenance of different UBC cell lines, and the relationship with their genomic signatures. They suggest that oxidative and non-oxidative metabolic states are primarily related to cell lines with low and high risk of progression, respectively. From this observation and our preliminary in vivo results, it appears that the lactate/alanine ratio in patients' urine is a good candidate to become a new marker to predict the conversion of low-grade tumors into more malignant forms.

scholarly journals A novel monoclonal antibody KMP1 has potential antitumor activity of bladder cancer by blocking CD44 in vivo and in vitro

2018 ◽  
Vol 7 (5) ◽  
pp. 2064-2077 ◽  
Author(s):  
Yujin Chen ◽  
Haifeng Wang ◽  
Yigang Zuo ◽  
Ning Li ◽  
Mingxia Ding ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Bladder Cancer ◽  
Antitumor Activity ◽  
Potential Antitumor

scholarly journals 878 Novel microbial immunotherapy approach for the treatment of bladder cancer

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A920-A920
Author(s):  
Nicholas Glanville ◽  
Tobi Oke ◽  
Sonia Domingos-Pereira ◽  
Lenka Polak ◽  
Denise Nardelli-Haefliger ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Bladder Cancer ◽  
Immune Responses ◽  
Median Survival ◽  
Standard Of Care ◽  
Significant Survival ◽  
Flow Cytometry Staining ◽  
Muscle Invasive

BackgroundMicrobial immunotherapy, in the form of intravesical Bacillus Calmette-Guérin (BCG), is the standard-of-care for non-muscle invasive bladder cancer (NMIBC). BCG therapy is associated with significant side-effects, high disease recurrence and progression rates, and product supply shortages, leaving a significant unmet medical need for bladder cancer patients.1 2 We sought to establish the preclinical safety and efficacy of live-attenuated Salmonella enterica Typhi strain ZH9 (ΔaroC, ΔssaV) as a novel microbial immunotherapy.MethodsTherapeutic efficacy of intravesical ZH9 was established in the murine orthotopic, syngeneic MB49 bladder tumor model. Tumor-bearing animals were treated with a single intravesical dose of ZH9 or OncoTice BCG and long-term survival comparisons were evaluated by log-rank (Mantel-Cox) test. ZH9 interaction with urothelial cancer cells was investigated using in vitro invasion assays and flow cytometry staining for intracellular Salmonella common antigen (CSA-1) and propidium iodide for cell death. Local immune responses were analyzed by flow cytometry staining of disaggregated mouse bladders.ResultsMice treated with a single intravesical dose of ZH9 2 days after MB49 tumor inoculation demonstrated significant survival benefit compared to vehicle treated (median survival 49.5 vs. 31 days, p=0.003) and BCG treated animals (median survival 49.5 vs. 27.5 days, p<0.001). A second, stringent model setup with intravesical treatment 4 days after tumor inoculation showed significant efficacy of ZH9 versus vehicle and BCG (median survival 30 vs. 20.5 (p=0.003) and 23.5 (p=0.025) days, respectively). Surviving ZH9 treated animals demonstrated 100% protection from tumor take following repeated intravesical challenge with MB49 tumor cells, suggesting lasting anti-tumor immunity resulting from ZH9 treatment. In vitro, intracellular flow cytometry in human (UMUC3, T24, RT4, 5637) and mouse (MB49) urothelial cancer cell lines showed that ZH9 invaded and induced cell death in all cell lines. In vivo, a single treatment with intravesical ZH9 resulted in strong cellular immune responses characterized by recruitment of monocytes, NK cells, CD4+ and CD8+ T cells, and dendritic cells with an activated, cross-presenting (Ly6C+, CD103+) phenotype. Intravesical ZH9 resulted in a greater magnitude and duration of immune cell recruitment in the urothelium compared to a single equivalent dose of intravesical BCG.ConclusionsLive-attenuated Salmonella strain ZH9 demonstrated a significant survival benefit over the standard-of-care OncoTice BCG in an orthotopic bladder cancer model. ZH9 demonstrated direct tumour cell killing in vitro and induction of robust cellular immune responses in vivo. Preclinical studies indicate significant therapeutic potential of intravesical ZH9 as a novel microbial immunotherapy in bladder cancer.ReferencesLiu Y, Lu J, Huang Y, Ma L. Clinical spectrum of complications induced by intravesical immunotherapy of Bacillus Calmette-Guérin for bladder cancer. J Oncol 2019 March 10;2019:6230409.vanRhijn BW, Burger M, Lotan Y, et al. Recurrence and progression of disease in non-muscle-invasive bladder cancer: from epidemiology to treatment strategy. Eur Urol 2009 September;56(3):430–42.Ethics ApprovalThe studies were conducted under approval from Pennsylvania State College of Medicine Institutional Animal Care and Use Committee approval number 47682, or with approval of the Cantonal Veterinary Office of Canton de Vaud, Switzerland.

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