Flores-Fernandez, Jose MiguelPesch, VerenaSriraman, AishwaryaChimal-Juarez, EnriqueAmidian, SaraWang, XiongyaoDuckering, CalebFang, AndrewReithofer, SaraMa, LiangCortez, Leonardo M.Sim, Valerie L.Tamgüney, GültekinWille, Holger2024-09-232024-09-232024-04-09Flores-Fernandez JM, Pesch V, Sriraman A, et al. Rational design of structure-based vaccines targeting misfolded alpha-synuclein conformers of Parkinson's disease and related disorders. Bioeng Transl Med. 2024;9(4):e10665. Published 2024 Apr 9. doi:10.1002/btm2.10665https://hdl.handle.net/1805/43490Synucleinopathies, including Parkinson's disease (PD), multiple system atrophy (MSA), and dementia with Lewy bodies (DLB), are neurodegenerative disorders caused by the accumulation of misfolded alpha-synuclein protein. Developing effective vaccines against synucleinopathies is challenging due to the difficulty of stimulating an immune-specific response against alpha-synuclein without causing harmful autoimmune reactions, selectively targeting only pathological forms of alpha-synuclein. Previous attempts using linear peptides and epitopes without control of the antigen structure failed in clinical trials. The immune system was unable to distinguish between native alpha-synuclein and its amyloid form. The prion domain of the fungal HET-s protein was selected as a scaffold to introduce select epitopes from the surface of alpha-synuclein fibrils. Four vaccine candidates were generated by introducing specific amino acid substitutions onto the surface of the scaffold protein. The approach successfully mimicked the stacking of the parallel in-register beta-sheet structure seen in alpha-synuclein fibrils. All vaccine candidates induced substantial levels of IgG antibodies that recognized pathological alpha-synuclein fibrils derived from a synucleinopathy mouse model. Furthermore, the antisera recognized pathological alpha-synuclein aggregates in brain lysates from patients who died from DLB, MSA, or PD, but did not recognize linear alpha-synuclein peptides. Our approach, based on the rational design of vaccines using the structure of alpha-synuclein amyloid fibrils and strict control over the exposed antigen structure used for immunization, as well as the ability to mimic aggregated alpha-synuclein, provides a promising avenue toward developing effective vaccines against alpha-synuclein fibrils.en-USAttribution 4.0 InternationalParkinson's diseaseBeta‐solenoid structureParallel in‐register beta‐sheet structureProtein engineeringRational designVaccineArticle