Towards Establishment of an Ex Vivo Bladder Tumor Model using Precision Cut Tumor Slices (PCTS) for Evaluation of Existing and Novel Treatments

Abstract

To continue the development of novel therapeutics to treat bladder cancer, a reliable model for preclinical testing of treatments and studying of the disease mechanism is essential. Ex vivo culturing of precision cut tumor slices (PCTS) provides a model that can retain the tumor architecture and its tumor microenvironment including the immune cell compartment, but data on PCTS based on bladder tumor is very limited. In this study, we partly establish a method for ex vivo culturing of PCTS from bladder tumors by examining the preservation over time in different media and further evaluating the secreted immune mediators and composition of the immune cell compartment upon treatment with immune cell activators and therapeutic agents.

Bladder tumors were obtained from treatment naïve patients undergoing tumor resection. The PCTS were generated by cutting the fresh tissue into 300 µm slices in a vibratome. Slices were ex vivo cultured on membrane inserts in 6-well plates. The first investigation included ex vivo culturing of slices in four different media for six days to evaluate the preservation of tissue structure and cell subsets. The second part included ex vivo culturing overnight followed by 24 h treatment with anti-CD3/CD28 antibodies, lipopolysaccharide, nivolumab, anti-CD40 antibodies or chemotherapeutics (cisplatin or gemcitabine).

PCTS were preserved sufficiently for three days, although some loss of immune cells was observed. Upon treatment, we observed an immunogenic response with all treatments, but for therapeutic agents not every patient responded, highlighting patient heterogeneity captured by the model. Ex vivo culturing of PCTS offers a promising method to evaluate novel therapeutics with analysis in multiplex immunoassay (Luminex) capturing the immunogenic response and multiplex flow cytometry complementing by identifying the different cell subsets and activation markers.

Popular Abstract

Establishment of a Bladder Cancer Model based on Fresh Tumor Slices for Evaluation of Existing and Novel Treatments

Cancer is a complex disease with almost everyone relating to a loss of someone dear, and despite impressive advancements in cancer research, some subtypes remain hard to treat. Bladder cancer is one of them, causing over 200,000 deaths annually, with an abnormally high risk of relapse, indicating that research progress is still needed. Current treatments for bladder cancer include surgery, chemotherapy and immunotherapy. While some patient responds well to the available treatments, not all patients benefit equally. To improve survival rates and reduce recurrence in bladder cancer, new drugs need to be developed.

Before these new drugs can reach patients suffering from bladder cancer, they need to be tested in biomedical models that mimic how the tumor behaves in the human body. Tumors consists of a diverse environment, referred to as the tumor microenvironment, including various cell types, e.g. immune cells, that communicate through tiny chemical messages. Many existing biomedical models fails to fully replicate this complexity, lacking the ability to include both cancer cells, immune cells and their interactions through small molecules. However, culturing slices of fresh tumor is a promising model for studying drug effects in bladder cancer, by its potential to mimic the tumor reality closely comparable to the patient-specific tumor environment present in humans.

In this project, we cut human bladder tumor tissue into thin slices of tumor by a specific precision cut technique. These slices are then placed in a nutrient-rich solution, where they can be treated with different pharmaceuticals. The aim of this project is to establish a bladder tumor model using these thin tumor slices to evaluate both existing and novel treatments. We show that culturing of tumor tissue can preserve the tumor’s structure for three days and that a treatment response can be induced and monitored upon addition of therapeutic drugs. The model also reflects the expected individual differences in patients’ tumors, making it a promising tool for preclinical testing of novel treatments.

• author: Hugoson, Jenny and Werner, Nadja
• supervisor: Kristina Lundberg; Sarah Richtmann
• organization: Department of Immunotechnology
• course: KIMM05 20241
• year: 2024
• type: H2 - Master's Degree (Two Years)
• subject: Biology and Life Sciences; Medicine and Health Sciences
• language: English
• id: 9163281
• date added to LUP: 2024-06-14 08:13:17
• date last changed: 2024-06-17 15:10:16