Differential seeding by exogenous R2 and R3 fibrils influences autophagic degradation of intracellular tau aggregates in Tau K18 P301S cells

Aggregation of misfolded tau protein is a common feature of tauopathies. Cells employ diverse mechanisms to eliminate misfolded tau with different conformations, contributing to the varied clinical and pathological manifestations of tauopathies. This study focuses on the clearance of seeded tau aggregates following the induction of biosensor cells with R2 and R3 fibrils that exhibit distinct aggregation kinetics and seeding potencies. Our hypothesis is that the dissimilarity in time-dependent intracellular seeding induced by R2 and R3 fibrils underlies the variation in autophagy failure. These discrepancies may account for the heterogeneity of pathology and disease progression between 3R and 4R tauopathies, given the absence of R2 in 3R tau isoforms. In R2-induced cells, alterations in p62 and LC3II/I levels, indicative of proteotoxic stress and autophagy failure, occur sooner than in R3-induced cells. Conversely, LAMP1 levels remained unaffected, suggesting a failure in the fusion of aggregate-containing autophagosomes with lysosomes. This autophagic failure may increase seed-dependent intracellular aggregation in induced cells. Consequently, we assessed the impact of autophagy inducers on the clearance of intracellular tau aggregates in induced cells. Epigallocatechin gallate (EGCG) demonstrated the highest efficacy in inducing autophagy and reducing p62 levels, decreasing seeding and clearing aggregates. Overall, this study elucidates the differential effects of prion-like R2 and R3 strains on autophagy and highlights how compounds like EGCG can selectively reduce tau aggregation to treat specific tauopathies. We provide insights into the distinct mechanisms of autophagy failure and autophagy clearance of intracellular aggregates in cells induced with R2 and R3 fibrils. ### Competing Interest Statement The authors have declared no competing interest.