Intracellular accumulation of tau inhibits autophagosome formation by activating TIA1-amino acid-mTORC1 signaling

Abstract

Background Autophagy dysfunction plays a crucial role in tau accumulation and neurodegeneration in Alzheimer’s disease (AD). This study aimed to investigate whether and how the accumulating tau may in turn affect autophagy. Methods The primary hippocampal neurons, N2a and HEK293T cells with tau overexpression were respectively starved and treated with vinblastine to study the effects of tau on the initiating steps of autophagy, which was analysed by Student's two-tailed t-test. The rapamycin and concanamycin A were employed to inhibit the mammalian target of rapamycin kinase complex 1 (mTORC1) activity and the vacuolar H+-ATPase (v-ATPase) activity, respectively, which were analysed by One‐way ANOVA with post hoc tests. The Western blotting, co-immunoprecipitation and immunofluorescence staining were conducted to gain insight into the mechanisms underlying the tau effects of mTORC1 signaling alterations, as analysed by Student's two-tailed t-test or One‐way ANOVA with post hoc tests. The autophagosome formation was detected by immunofluorescence staining and transmission electron microscopy. The amino acids (AA) levels were detected by high performance liquid chromatography (HPLC). Results We observed that overexpressing human full-length wild-type tau to mimic AD-like tau accumulation induced autophagy deficits. Further studies revealed that the increased tau could bind to the prion-related domain of T cell intracellular antigen 1 (PRD-TIA1) and this association significantly increased the intercellular level of amino acids (Leucine, P = 0.0038; Glutamic acid, P = 0.0348; Alanine, P = 0.0037; Glycine, P = 0.0104), with concordant upregulation of mTORC1 activity [phosphorylated eukaryotic translation initiation factor 4E-binding protein 1 (p-4EBP1), P < 0.0001; phosphorylated 70 kDa ribosomal protein S6 kinase 1 (p-p70S6K1), P = 0.0001, phosphorylated unc-51-like autophagy-activating kinase 1 (p-ULK1), P = 0.0015] and inhibition of autophagosome formation [microtubule-associated protein light chain 3 II (LC3 II), P = 0.0073; LC3 puncta, P < 0.0001]. As expected, this tau-induced deficit of autophagosome formation in turn aggravated tau accumulation. Importantly, we also found that blocking TIA1 and tau interaction by overexpressing PRD-TIA1, downregulating the endogenous TIA1 expression by shRNA, or downregulating tau protein level by a small proteolysis targeting chimera (PROTAC) could remarkably attenuate tau-induced autophagy impairment. Conclusions Our findings reveal that AD-like tau accumulation inhibits autophagosome formation and induces autophagy deficits by activating the TIA1/amino acid/mTORC1 pathway, and thus this work reveals new insight into tau-associated neurodegeneration and provides evidence supporting the use of new therapeutic targets for AD treatment and that of related tauopathies.

The Role of Tau Protein on Alzheimer’s Disease

“…This means tau overexpression triggered the dysfunction of autophagy. At the same time, the LC3 II protein level was also decreased, which was measured by immunofluorescence staining [8]. In primary hippocampal neuron cells with Lenti-eGFP-P2A-tau overexpression, a decline of autophagosome formation was witnessed [8].…”

“…LC3 acts as the autophagy marker. Tau5, which is an antibody of tau proteins, firstly detected an increment in its level, indicating that there existed overexpression of tau [8]. This means tau overexpression triggered the dysfunction of autophagy.…”

“…At the same time, the LC3 II protein level was also decreased, which was measured by immunofluorescence staining [8]. In primary hippocampal neuron cells with Lenti-eGFP-P2A-tau overexpression, a decline of autophagosome formation was witnessed [8]. This is because of the decrease of two components, autophagy-activating kinase 1 (ULK1) and autophagy protein 13 (ATG13), which are responsible for making autophagosomes [9].…”

“…This is because of the decrease of two components, autophagy-activating kinase 1 (ULK1) and autophagy protein 13 (ATG13), which are responsible for making autophagosomes [9]. Furthermore, the mammalian target of rapamycin kinase complex 1 (mTORC1) functions in a vital upstream pathway of autophagy, whose activity can be regulated by tau overexpression [8]. Tau overexpression upregulates the amino acid pathway by increasing the levels of leucine, glutamic acid, alanine, and glycine, which continuously raises the level of anti-lysosome-associated membrane glycoprotein 1(LAMP1) and mammalian target of rapamycin (mTOR) [10], by which the mTORC1 pathway can be activated, and autophagosome formation decreased remarkably [10].…”

“…To be more specific, tau could bind T cell intracellular antigen 1 (TIA1) with the N-terminal, thus an increased TIA1 level could be seen in the cytoplasm under the effect of tau overexpression, which could lead to the activation of amino acid and mTORC1 pathways [9]. The complement of the prion-related domain of T cell intracellular antigen 1 (TIA1-PRD) can mitigate the dysfunction of autophagy triggered by overexpression of tau [8].…”