CRISPR-Cas9 Genome Editing Using Lipid Nanoparticles for Mantle Cell Lymphoma Therapy

Mantle Cell Lymphoma (MCL) is an aggressive rare B-cell malignancy, genetically characterized by the t(11;14) translocation that leads to overexpression of CyclinD1. Despite advances in chemotherapy and immunotherapy, all patients eventually relapse, rendering the disease incurable with a short median survival of 7-8 years. Therefore, new therapeutic strategies are most needed. SOX11 has emerged as a key transcription factor in the pathogenesis of MCL as it is highly expressed in ~90% of patients, while absent in normal B-cells. Its specific expression in MCL, particularly in aggressive forms, suggests that SOX11 may serve as a potential therapeutic target. CRISPR/Cas9 is a powerful tool for specific gene editing that holds great promise in treating various conditions, including hematological malignancies. We aim to harness CRISPR/Cas9 technology to specifically knockout SOX11 as a novel potential therapy for MCL. For this, we will generate lipid nanoparticles (LNPs) co-encapsulating Cas9 mRNA with sgRNA, as a safe and efficient delivery platform. Our results show that SOX11 knockdown in MCL cell lines leads to their significant cell death. For the utilization of CRISPR/Cas9, we screened for multiple sgRNAs and found three efficient candidates that lead to ~70% editing. We are now screening LNP formulations for efficient delivery to MCL cells. For in-vivo efficacy evaluation of SOX11-CRISPR-LNPs, we established a new xenograft model that exhibits high engraftment levels of MCL cells in the bone marrow, as in most MCL cases. Our potential ability to disrupt gene expression in MCL may open new avenues for treating this devastating disease.