Baltissen Danny, Bold Charlotte S, Rehra Lena, Banićević Marija, Fricke Justus, Just Jennifer, Ludewig Susann, Buchholz Christian J, Korte Martin, Müller Ulrike C
Department of Functional Genomics, Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany.
Department of Cellular Neurobiology, Zoological Institute, Technical University of Braunschweig, Braunschweig, Germany.
Front Cell Neurosci. 2023 Jan 26;17:1106176. doi: 10.3389/fncel.2023.1106176. eCollection 2023.
The Tau protein can be phosphorylated by numerous kinases. In Alzheimer's disease (AD) hyperphosphorylated Tau species accumulate as neurofibrillary tangles that constitute a major hallmark of AD. AD is further characterized by extracellular Aβ plaques, derived from the β-amyloid precursor protein APP. Whereas Aβ is produced by amyloidogenic APP processing, APP processing along the competing non-amyloidogenic pathway results in the secretion of neurotrophic and synaptotrophic APPsα. Recently, we demonstrated that APPsα has therapeutic effects in transgenic AD model mice and rescues Aβ-dependent impairments. Here, we examined the potential of APPsα to regulate two major Tau kinases, GSK3β and CDK5 in THY-Tau22 mice, a widely used mouse model of tauopathy. Immunohistochemistry revealed a dramatic increase in pathologically phosphorylated (AT8 and AT180) or misfolded Tau species (MC1) in the hippocampus of THY-Tau22 mice between 3 and 12 months of age. Using a highly sensitive radioactive kinase assay with recombinant human Tau as a substrate and immunoblotting, we demonstrate an increase in GSK3β and CDK5 activity in the hippocampus of THY-Tau22 mice. Interestingly, AAV-mediated intracranial expression of APPsα in THY-Tau22 mice efficiently restored normal GSK3β and CDK5 activity. Western blot analysis revealed upregulation of the CDK5 regulatory proteins p35 and p25, indicating CDK5 hyperactivation in THY-Tau22 mice. Strikingly, AAV-APPsα rescued p25 upregulation to wild-type levels even at stages of advanced Tau pathology. Sarkosyl fractionation used to study the abundance of soluble and insoluble phospho-Tau species revealed increased soluble AT8-Tau and decreased insoluble AT100-Tau species upon AAV-APPsα injection. Moreover, AAV-APPsα reduced misfolded (MC1) Tau species, particularly in somatodendritic compartments of CA1 pyramidal neurons. Finally, we show that AAV-APPsα upregulated PSD95 expression and rescued deficits in spine density of THY-Tau22 mice. Together our findings suggest that APPsα holds therapeutic potential to mitigate Tau-induced pathology.
Tau蛋白可被多种激酶磷酸化。在阿尔茨海默病(AD)中,过度磷酸化的Tau蛋白以神经原纤维缠结的形式积累,这是AD的一个主要标志。AD的另一个特征是细胞外Aβ斑块,它源自β-淀粉样前体蛋白APP。Aβ是由淀粉样生成性APP加工产生的,而APP沿竞争性非淀粉样生成途径进行加工则导致神经营养性和突触营养性APPsα的分泌。最近,我们证明APPsα在转基因AD模型小鼠中具有治疗作用,并能挽救Aβ依赖性损伤。在此,我们研究了APPsα在THY-Tau22小鼠(一种广泛使用的tau蛋白病小鼠模型)中调节两种主要Tau激酶GSK3β和CDK5的潜力。免疫组织化学显示,在3至12月龄的THY-Tau22小鼠海马中,病理磷酸化(AT8和AT180)或错误折叠的Tau蛋白(MC1)显著增加。使用以重组人Tau为底物的高灵敏度放射性激酶测定法和免疫印迹法,我们证明THY-Tau22小鼠海马中GSK3β和CDK5活性增加。有趣的是,腺相关病毒(AAV)介导的APPsα在THY-Tau22小鼠颅内表达可有效恢复正常的GSK3β和CDK5活性。蛋白质印迹分析显示CDK5调节蛋白p35和p25上调,表明THY-Tau22小鼠中CDK5过度激活。令人惊讶的是,即使在Tau病理晚期阶段,AAV-APPsα也能将p25上调挽救至野生型水平。用于研究可溶性和不溶性磷酸化Tau蛋白丰度的 Sarkosyl分级分离显示,注射AAV-APPsα后,可溶性AT8-Tau增加,不溶性AT100-Tau减少。此外,AAV-APPsα减少了错误折叠的(MC1)Tau蛋白,特别是在CA1锥体神经元的树突-胞体区室。最后,我们表明AAV-APPsα上调了PSD95表达,并挽救了THY-Tau22小鼠的脊柱密度缺陷。我们的研究结果共同表明,APPsα具有减轻Tau诱导的病理变化的治疗潜力。