Customized antisense oligonucleotide-based therapy for neurofilament-associated Charcot-Marie-Tooth disease

Abstract DNA-based therapeutics have emerged as a revolutionary approach for addressing the treatment gap in rare inherited conditions by targeting the fundamental genetic causes of disease. Charcot–Marie–Tooth (CMT) disease, a group of inherited neuropathies, represents one of the most prevalent Mendelian disease groups in neurology and is characterized by diverse genetic aetiology. Axonal forms of CMT, known as CMT2, are caused by dominant mutations in >30 different genes that lead to degeneration of lower motor neuron axons. Recent advances in antisense oligonucleotide therapeutics have shown promise in targeting neurodegenerative disorders. Here, we elucidate pathomechanistic changes contributing to variant specific molecular phenotypes in CMT2E, caused by a single nucleotide substitution (p.N98S) in the neurofilament light chain gene (NEFL). We used a patient-derived induced pluripotent stem cell-induced motor neuron model that recapitulates several cellular and biomarker phenotypes associated with CMT2E. Using an antisense oligonucleotide treatment strategy targeting a heterozygous gain-of-function variant, we aimed to resolve molecular phenotypic changes observed in the CMT2E p.N98S subtype. To determine the therapeutic potential of antisense oligonucleotide, we applied our treatment strategy in induced pluripotent stem cell-derived motor neurons and used both established and new biomarkers of peripheral nervous system axonal degeneration. Our findings demonstrated a significant decrease in clinically relevant biomarkers of axonal degeneration, presenting the first clinically viable genetic therapeutic for CMT2E. Similar strategies could be used to develop precision medicine approaches for otherwise untreatable gain-of-function inherited disorders.