RT-QuIC Using C-Terminally Truncated α-Synuclein Forms Detects Differences in Seeding Propensity of Different Brain Regions from Synucleinopathies

Aggregated alpha-synuclein (alpha Syn) protein is a core pathological feature of Parkinson's disease (PD) and dementia with Lewy bodies (DLB). Both PD and DLB demonstrate the presence of diverse intracellular alpha-synuclein (alpha Syn) species, including C-terminally truncated alpha Syn (C-alpha Syn), although it is unknown how C-alpha Syn species contribute to disease progression. Using recombinant C-alpha Syn and PD and DLB brain lysates as seeds in the real-time quaking-induced conversion (RT-QuIC) assay, we explored how C-alpha Syn may be involved in disease stratification. Comparing the seeding activity of aqueous-soluble fractions to detergent-soluble fractions, and using alpha Syn 1-130 as substrate for the RT-QuIC assay, the temporal cortex seeds differentiated PD and DLB from healthy controls. In contrast to the temporal cortex, where PD and DLB could not be distinguished, alpha Syn 1-130 seeded by the detergent-soluble fractions from the PD frontal cortex demonstrated greater seeding efficiency compared to the DLB frontal cortex. Moreover, proteinase K-resistant (PKres) fragments from the RT-QuIC end products using C-alpha Syn 1-130 or C-alpha Syn 1-115 were more obvious in the frontal cortex compared to the temporal cortex. Morphological examinations of RT-QuIC end products showed differences in the size of the fibrils between C-alpha Syn 1-130 and C-alpha Syn 1-115, in agreement with the RT-QuIC results. These data show that C-alpha Syn species can distinguish PD from DLB and suggest diversity in alpha Syn species across these synucleinopathies, which could play a role in disease progression.