Extracellular Vesicle-Enhanced Mitochondrial Therapy for ALS and Neurodegeneration - Project Summary Neurodegenerative diseases (NDs), such as amyotrophic lateral sclerosis (ALS), Alzheimer’s disease, and Parkinson’s disease, are characterized by progressive loss of nerve cells (neurons) in the brain and spinal cord, leading to severe decline in movement, memory and cognition. Despite their significant societal and personal impact, effective treatments remain elusive mainly due to the complexity of disease mechanisms and the challenge of delivering therapeutics to affected central nervous system (CNS). Mitochondrial dysfunction is a central driver of neuronal degeneration, and mitochondrial transplantation, introducing healthy mitochondria into damaged cells, has shown therapeutic promise in preclinical ND models. However, its translational application is limited by inefficient CNS delivery, resulting from poor penetration of the blood brain barrier (BBB), mitochondrial instability, and limited cellular uptake. To address these limitations, we propose to develop a novel extracellular vesicle (EV)-based platform for mitochondrial delivery. EVs are natural intercellular communicators, capable of crossing cell membranes and the BBB to deliver bioactive molecules, such as nucleic acids, protein, and even organelles like mitochondria, between cells. Notably, EVs derived from neural progenitor cells (NPC- EVs) possess intrinsic neuroprotective and regenerative properties inherited from parent cells, mitigating neuronal death and disease progression in preclinical models of NDs. Our preliminary data show that co- incubation of EVs with isolated mitochondria significantly enhances mitochondrial uptake by neural cells in vitro and brain accumulation in vivo following non-invasive intranasal administration. Building on these findings, we hypothesize that NPC-EV-based mitochondrial delivery will improve CNS targeting and therapeutic efficacy by combining the bioenergetic benefits of mitochondrial transplantation with the intrinsic delivery and therapeutic properties of NPC-EVs. In the proposed study, we will (1) enhance the association of mitochondria with NPC- EVs via surface modification to improve delivery to the CNS, and (2) evaluate the therapeutic efficacy of NPC- EV-mediated mitochondrial delivery in established cellular and mouse models of ALS. Successful completion of this study will establish proof-of-concept for NPC-EV-based mitochondrial therapy aimed at restoring bioenergetics in damaged nerve cells and rescuing neuronal viability and function in ALS and potentially other NDs. By overcoming key delivery challenges in ND treatment, this strategy combines the therapeutic benefits of both mitochondrial transplantation and NPC-EVs, holding strong promise to advance the clinical translation of mitochondrial therapies.
