Lipid nanoparticle formulation for gene editing and RNA-based therapies for glioblastoma

Yanhong Zhang; Rosalia Rabinovsky; Evgeny Deforzh; Ami Kobayashi; Anastasia Kuzkina; Johnna Francis Varghese; Damita Rai; Joanna A Korecka; Vikram Khurana; Gopal Murugaiyan; David Morrissey; Erik J Uhlmann; Anna M Krichevsky

doi:10.1093/neuonc/noaf162

Lipid nanoparticle formulation for gene editing and RNA-based therapies for glioblastoma

Neuro Oncol. 2025 Jul 11:noaf162. doi: 10.1093/neuonc/noaf162. Online ahead of print.

Affiliations

¹ Ann Romney Center for Neurologic Diseases, Department of Neurology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115 USA.
² Division of Movement Disorders, Department of Neurology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115 USA.
³ Jumble Therapeutics, Cambridge, MA 02139 USA.

PMID: 40653819
DOI: 10.1093/neuonc/noaf162

Abstract

Background: Glioblastoma (GBM), one of the deadliest cancers, resists current therapies, with drug development hindered by its high heterogeneity. However, GBM consistently relies on microRNA-10b (miR-10b), a key driver of glioma growth and a promising therapeutic target. miR-10b gene editing represents a potential treatment, but effective delivery strategies for gene editing systems in GBM remain unexplored.

Methods: We developed lipid nanoparticles (LNPs) encapsulating Cas9 mRNA and a miR-10b-targeting sgRNA (termed miRTEN). miRTEN was tested in glioma stem cells (GSCs) and orthotopic GBM models to assess therapeutic efficacy, immune responses, and safety.

Results: Intracerebroventricular (ICV) injections of miRTEN enabled broad and durable Cas9 mRNA expression and miR-10b gene editing in tumor core and invasive areas across diverse GBM models. miRTEN significantly suppressed tumor growth, reduced GSC proliferation and viability, with therapeutic outcomes correlating with dose-dependent miR-10b suppression. Combining miRTEN with temozolomide (TMZ) further enhanced tumor suppression, overcoming TMZ resistance and improving survival. In immunocompetent models, miRTEN activated anti-tumor immune responses, increased cytotoxic CD8+ T cells infiltration, and promoted durable immune memory, enabling tumor rejection upon rechallenge. Safety assessments demonstrated that miRTEN selectively targets GBM cells, sparing normal brain tissues and causing no significant off-target toxicity.

Conclusion: As in vivo CRISPR-based drugs advance toward clinical applications, our findings demonstrate the potential of LNPs-mediated CRISPR-Cas9 systems for targeted miR-10b editing and, more generally, gene editing and RNA therapies for GBM. miRTEN monotherapy, as well as its combination with standard care, offers a promising, safe, and effective approach to improving outcomes in GBM.

Keywords: CRISPR-Cas9; Glioblastoma; gene editing; lipid nanoparticles; microRNA-10b.