Development of an RNA Aptamer as a Therapeutic Agent for Synucleinopathies

Abstract

The aggregation of -synuclein (Syn), a 140-mer protein, has been implicated in the pathogenesis of Parkinsons disease, multiple system atrophy, and dementia with Lewy bodies. KTKEGV repeats (KR) of Syn are key mediators of prion-like propagation and neurodegeneration. Despite the availability of symptomatic treatments, no current therapy effectively delays disease progression. Here we report a 77-nucleotide (nt) RNA aptamer (1R6) with potent affinity and selectivity for Syn1-95 (KD = 18 nM) through in vitro selection. 1R6 significantly inhibited Syn oligomerization and {beta}-sheet-rich fibril assembly and promoted disaggregation of preformed fibrils. Additionally, 1R6 suppressed Syn seeding, as determined by FRET-based cellular biosensor cell assay. Cellular studies revealed that 1R6 cotransfection completely prevented Syn-induced cytotoxicity. To assess the protective effects of 1R6 in vivo, we used a Drosophila melanogaster model expressing human Syn in neurons. Flies fed with 1R6 showed improved locomotor defects, reduced photoreceptor degeneration, and decreased Syn levels in the head. Structural characterization through 1H-15N heteronuclear multiple quantum correlation nuclear magnetic resonance experiments demonstrated that 1R6 targets KR motifs, a finding further supported by in silico simulations. Our findings indicate that RNA aptamers, such as 1R6, may represent promising therapeutic candidates for synucleinopathies, thus opening new avenues in the treatment of these diseases.