An Innovative Mitochondrial-targeted Gene Therapy for Cancer Treatment

Abstract

Targeting cancer cell mitochondria holds great promise for cancer treatment, yet current strategies for specifically and effectively destroying cancer mitochondria in vivo are limited. Here, we introduce mLumiOpto, an innovative mitochondrial-targeted luminoptogenetics technology designed to directly disrupt the inner mitochondrial membrane (IMM) potential and induce cancer cell death. We synthesize a blue light-gated channelrhodopsin (CoChR) in the IMM with a mitochondrial leading sequence and co-express a blue bioluminescence-emitting Nanoluciferase (NLuc) in the cytosol of the same cells. The mLumiOpto genes are selectively delivered to cancer cells in vivo by using two strategies: adeno-associated virus (AAV) carrying a cancer-specific promoter (cfos) or cancer-targeted monoclonal antibody-tagged exosome-associated AAV (mAb-Exo-AAV). Induction with ViviRen (an NLuc luciferin) elicits robust endogenous NLuc bioluminescence, which activates mitochondrial CoChR, triggering cancer cell IMM permeability disruption, IMM potential depolarization and subsequent cell death. Importantly, mLumiOpto demonstrates remarkable efficacy in reducing tumor burden and killing tumor cells in glioblastomas (administrated with AAV) or triple-negative breast cancers (administrated with mAb-Exo-AAV) xenografted mouse models. These findings establish mLumiOpto as a novel and promising therapeutic strategy by targeting cancer cell mitochondria in vivo.