Intracerebral Administration of a Ligand-ASO Conjugate Selectively Reduces alpha-Synuclein Accumulation in Monoamine Neurons of Double Mutant Human A30P*A53T*alpha-Synuclein Transgenic Mice

alpha-Synuclein (alpha-Syn) protein is involved in the pathogenesis of Parkinson's disease (PD). Point mutations and multiplications of the alpha-Syn, which encodes the SNCA gene, are correlated with early-onset PD, therefore the reduction in a-Syn synthesis could be a potential therapy for PD if delivered to the key affected neurons. Several experimental strategies for PD have been developed in recent years using oligonucleotide therapeutics. However, some of them have failed or even caused neuronal toxicity. One limiting step in the success of oligonucleotide-based therapeutics is their delivery to the brain compartment, and once there, to selected neuronal populations. Previously, we developed an indatraline-conjugated antisense oligonucleotide (IND-1233-ASO), that selectively reduces alpha-Syn synthesis in midbrain monoamine neurons of mice, and nonhuman primates. Here, we extended these observations using a transgenic male mouse strain carrying both A30P and A53T mutant human alpha-Syn (A30P*A53T*alpha-Syn). We found that A30P*A53T*alpha-Syn mice at 4-5 months of age showed 3.5-fold increases in human alpha-Syn expression in dopamine (DA) and norepinephrine (NE) neurons of the substantia nigra pars compacta (SNc) and locus coeruleus (LC), respectively, compared with mouse alpha-Syn levels. In parallel, transgenic mice exhibited altered nigrostriatal DA neurotransmission, motor alterations, and an anxiety-like phenotype. Intracerebroventricular IND-1233-ASO administration (100 mu g/day, 28 days) prevented the alpha-Syn synthesis and accumulation in the SNc and LC, and recovered DA neurotransmission, although it did not reverse the behavioral phenotype. Therefore, the present therapeutic strategy based on a conjugated ASO could be used for the selective inhibition of alpha-Syn expression in PD-vulnerable monoamine neurons, showing the benefit of the optimization of ASO molecules as a disease modifying therapy for PD and related alpha-synucleinopathies.