Topical and systemic immunoreaction triggered by intravesical chemotherapy in an N-butyl-N-(4-hydroxybutyl) nitorosamine induced bladder cancer mouse model

Abstract

Intravesical bacillus Calmette-Guerin (BCG) treatment is the most common therapy to prevent progression and recurrence of non-muscle invasive bladder cancer (NMIBC). Although the immunoreaction elicited by BCG treatment is well documented, those induced by intravesical treatment with chemotherapeutic agents are much less known. We investigated the immunological profiles caused by mitomycin C, gemcitabine, adriamycin and docetaxel in the N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced orthotopic bladder cancer mouse model. Ninety mice bearing orthotopic bladder cancer induced by BBN were randomly divided into six groups and treated with chemotherapeutic agents once a week for four weeks. After last treatment, bladder and serum samples were analyzed for cell surface and immunological markers (CD4, CD8, CD56, CD204, Foxp3, and PD-L1) using immunohistochemistry staining. Serum and urine cytokine levels were evaluated by ELISA. All chemotherapeutic agents presented anti-tumor properties similar to those of BCG. These included changes in immune cells that resulted in fewer M2 macrophages and regulatory T cells around tumors. This result was compatible with those in human samples. Intravesical chemotherapy also induced systemic changes in cytokines, especially urinary interleukin (IL)-17A and granulocyte colony stimulating factor (G-CSF), as well as in the distribution of blood neutrophils, lymphocytes, and monocytes. Our findings suggest that intravesical treatment with mitomycin C and adriamycin suppresses protumoral immunity while enhancing anti-tumor immunity, possibly through the action of specific cytokines. A better understanding of the immunoreaction induced by chemotherapeutic agents can lead to improved outcomes and fewer side effects in intravesical chemotherapy against NMIBC.

Supplementary granulocyte macrophage colony‐stimulating factor to chemotherapy and programmed death‐ligand 1 blockade decreases local recurrence after surgery in bladder cancer

“…Serum was collected by allowing whole blood to clot at room temperature for 30 minutes, followed by centrifugation at 1000 g for 20 minutes at 4°C. ELISA was carried out as previously described . Details of commercially available kits for TGF‐β, interleukin (IL)‐6, and IL‐10 are listed in Table .…”

Intravesical treatment of chemotherapeutic agents sensitizes bacillus Calmette‑Guerin by the modulation of the tumor immune environment

“…To treat mice bearing BBN-induced bladder cancer with intravesical instillation, the following agents, which are commonly used in a clinical setting for NMIBC, were prepared: BCG (Nippon Kayaku, Tokyo, Japan), MMC (Kyowa Hakko Kirin, Tokyo, Japan) and ADM (Wako, Osaka, Japan). Each agent was diluted in sterile saline solution according to the manufacturer's instructions, and the dosage of each agent was determined by our previous study (23). Sterile phosphate-buffered saline (PBS) was used as a control.…”

“…The presence of TAM and Treg in the tumor microenvironment led to a poor response to the treatment with BCG (22). In an animal model, intravesical treatment with MMC and ADM also caused a reduction of TAMs and Tregs in the tumor microenvironment and induced systemic changes to cytokines, including interleukin (IL)-17 and granulocyte-colony stimulating factor (G-CSF) (23). There are limited studies describing details regarding immunoreactions induced by treatments with BCG and chemotherapeutic agents, and their efficacy from an immunological standpoint.…”

Cryptotanshinone Inhibites Bladder Cancer Cell Proliferation and Promotes Apoptosis via the PTEN/PI3K/AKT Pathway

“…Bladder cancer is the 7th most common tumour worldwide in males, while it is the 17th most common tumour in females and is one of the most deadly urothelial malignancies [1]. Almost three-quarters of newly diagnosed bladder cancer cases are non-muscle-invasive bladder cancer (NMIBC), which can be treated with transurethral resection of the tumours followed by intravesical instillation of chemotherapeutic drugs [2]. However, bladder cancers have a high recurrence rate, and it is estimated that 25% of patients with NMIBC develop muscle-invasive bladder carcinoma following treatment [3].…”