Unveiling the potential of electroporation-based glioblastoma treatment using patient-derived biomimetic models

Glioblastoma (GB) is the most aggressive type of brain tumors, with current standard treatments achieving a median survival of merely ~15 months. The blood-brain barrier substantially limits drug delivery, while therapy-resistant GB stem cells and immunosuppressive tumor microenvironment contribute to treatment failure. New therapeutic approaches are greatly needed.

Electroporation has emerged as a promising treatment modality that uses short, high-intensity electric pulses to either transiently increase cell membrane permeability for enhanced drug delivery (electrochemotherapy) or achieve non-thermal tumor ablation (irreversible electroporation). Encouraging results have been demonstrated in animal studies: irreversible electroporation achieved long-term tumor-free survival in canine patients with spontaneous gliomas, while electrochemotherapy showed complete tumor elimination in glioma-bearing rats with minimal side effects. However, neither canine nor rodent gliomas fully replicate the complexity of human GB. Before advancing to clinical trials, it is crucial to understand how electroporation affects human GB cells and their tumor microenvironment.

This project aims to systematically investigate electroporation effects using state-of-the-art biomimetic models of increasing complexity: multicellular spheroids from patient-derived GB cultures, organoids from human GB tissue, and their cocultures with immune cells. We will assess treatment effects on tumor cell viability, proliferation and invasion, while carefully evaluating responses of therapy-resistant GB stem cells, immune cells, and surrounding healthy tissue. The investigations will cover both irreversible electroporation and electrochemotherapy, as well as their combinations with standard treatments.

Our approach introduces three major innovations in electroporation research. First, we will use patient-derived primary GB cells which, unlike established cell lines, much more faithfully reproduce key aspects of tumor biology. Second, we will employ 3D multicellular spheroids that better represent the microscopic electric field distribution in tumors and model hindered diffusion of therapeutic agents. Third, we will pioneer the use of patient-derived GB organoids in electroporation research. These unique 3D structures closely resemble human tumors by retaining the complex heterogeneous cellular composition of the tumor microenvironment.

The project addresses fundamental questions that cannot be systematically investigated in vivo due to ethical constraints and experimental complexity. Our methodological approach enables unprecedented investigation of how surviving tumor cells respond to reversible electroporation, how cellular heterogeneity influences treatment efficacy, and how the immune system responds to the treatment. The insights gained will be essential for optimizing treatment protocols and identifying the most promising therapeutic combinations for clinical translation. While primarily focused on GB treatment, our findings will also benefit other promising applications of brain electroporation, such as non-thermal ablation of epileptic zones.

The project's feasibility is ensured through collaboration between two globally leading research groups. The group at the University of Ljubljana's Faculty of Electrical Engineering brings extensive expertise in electroporation research, treatment optimization, and clinical translation, while the group at the National Institute of Biology specializes in cancer biology and GB research and has developed unique resources including patient-derived GB cultures, spheroids from these primary cultures and GB organoids. This synergistic partnership ensures that technological developments will be guided by biological insights while biological findings will be interpreted within the context of physical principles, fostering scientific excellence and augmenting the translational aspect of the project.