Induction of immunogenic cell death by a DNA sensor system recognizing a cancer-specific genomic signature

Scientific background and problem identification. Immunogenic cell death is a form of regulated cell death that can initiate innate and adaptive immune response in immunocompetent hosts. It has been exploited for improving the efficiency of cancer treatments since the immune responses elicited by treatment-driven immunogenic cell death reinforce the therapeutic effects of conventional anticancer chemotherapy and radiotherapy. Although the data suggest that harnessing immunogenic cell death could be an effective treatment approach, these studies also underline the risk of its activation in normal tissues, which can lead to damage. Therefore, we propose to develop a DNA sensor system recognizing a cancer-specific genomic signature for induction of immunogenic cell death. Objectives of the study. We will focus on pyroptosis, a type of immunogenic cell death that results in the loss of integrity of plasma membrane due to activation of multimeric cytosolic protein complexes, i.e. inflammasomes. We propose to engineer late stages of pyroptosis based on the recent scientific advances in immunology and synthetic biology, to provide better control, efficient immune stimulation, and increased specificity of cancer immunotherapy. We aim to: 1. Establish a DNA sensor system for recognition of a cancer-specific DNA signature. By using CRISPR/Cas technology, we propose to establish a DNA sensor system that will enable cancer-specific DNA signature recognition. In scope of this project, we propose to target the BCR/ABL1 chromosomal translocation that is present in 95% of chronic myeloid leukemia cases. 2. Develop an effector and a signal amplification cascade for induction of immunogenic cell death. To induce immunogenic cell death, we propose to engineer the main effector of pyroptosis, i.e. gasdermin D, which would be cleaved after docking of the DNA sensor to target DNA, resulting in pore formation and consequent cell death. To ensure a sufficient gasdermin D activation that will result in cell death, we propose to develop a proteolysis-based DNA sensor system signal amplification cascade. 3. Test the specificity and efficiency of the DNA sensor system on 2D and 3D multicellular in vitro models. Interactions between cancer cells and the rest of the cells in the bone marrow microenvironment regulate various properties of leukemic cells. Hence, we propose to develop 2D and 3D multicellular in vitro models to assess the specificity and efficiency of the DNA sensor system. Relevance and potential impact of the results. Combining novel genome editing techniques and synthetic biology, the project aims to provide an original and innovative approach for cancer immunotherapy. The distinctive added value of this approach is in precise targeting, which would result in increased specificity and could potentially be used to improve current therapeutic approaches.