Genome Editing Using Lipid Nanoparticles for Mantle Cell Lymphoma Therapy

Mantle cell lymphoma (MCL) is an aggressive lymphoid neoplasm, characterized by abnormal proliferation of mature B-cells. Despite advances in chemotherapy and immunotherapy, all patients eventually relapse, rendering MCL incurable with current therapies. Therefore, new therapeutic strategies are 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 aggressive MCL suggests that SOX11 may serve as a potential therapeutic target. We aim to harness CRISPR-Cas9 technology to specifically knockout SOX11 as a novel potential therapy for MCL. For this, we generated targeted lipid nanoparticles (tLNPs) co-encapsulating Cas9 mRNA with sgRNA. Our results show that SOX11 knockdown in MCL cell lines leads to significant cell death. We screened multiple sgRNAs and found an efficient candidate reaching ~80% editing in MCL cells. Next, we screened and optimized different LNP formulations, identifying one that efficiently delivered CRISPR-Cas9 RNA into ~92% of the treated cells. To enhance specificity, we developed an anti-CD38 lipidated scFv, which was then incorporated into the LNPs, enabling selective binding and internalization. To evaluate the therapeutic effect of SOX11-CRISPR-LNPs in-vivo, we established a novel xenograft MCL model that exhibits high engraftment levels of MCL cells in the bone marrow, as in most MCL cases. Lastly, we performed biodistribution evaluations of the leading formulations, reaching the bone marrow of the MCL-bearing mice. Our potential ability to disrupt gene expression in MCL may open new avenues for treating this devastating disease.