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Hyperphosphorylation Renders Tau Prone to Aggregate and to Cause Cell Death

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Abstract

Alzheimer’s disease (AD) is a neurodegenerative disorder without a cure or prevention to date. Hyperphosphorylated tau forms the neurofibrillary tangles (NFTs) that correlate well with the progression of cognitive impairments. Animal studies demonstrated the pathogenic role of hyperphosphorylated tau. Understanding how abnormal phosphorylation renders a normal tau prone to form toxic fibrils is key to delineating molecular pathology and to developing efficacious drugs for AD. Production of a tau bearing the disease-relevant hyperphosphorylation and molecular characters is a pivotal step. Here, we report the preparation and characterization of a recombinant hyperphosphorylated tau (p-tau) with strong relevance to disease. P-tau generated by the PIMAX approach resulted in phosphorylation at multiple epitopes linked to the progression of AD neuropathology. In stark contrast to unmodified tau that required an aggregation inducer, and which had minimal effects on cell functions, p-tau formed inducer-free fibrils that triggered a spike of mitochondrial superoxide, induced apoptosis, and caused cell death at sub-micromolar concentrations. P-tau-induced apoptosis was suppressed by inhibitors for reactive oxygen species. Hyperphosphorylation apparently caused rapid formation of a disease-related conformation. In both aggregation and cytotoxicity, p-tau exhibited seeding activities that converted the unmodified tau into a cytotoxic species with an increased propensity for fibrillization. These characters of p-tau are consistent with the emerging view that hyperphosphorylation causes tau to become an aggregation-prone and cytotoxic species that underlies diffusible pathology in AD and other tauopathies. Our results further suggest that p-tau affords a feasible tool for Alzheimer’s disease mechanistic and drug discovery studies.

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Acknowledgments

We are grateful for the generous supply of MC-1 monoclonal antibodies by Peter Davies, and technical help and suggestions from Justin McCormick, Doug Gage, David DeWitt, John LaPres, Lisa Lapidus, Andrea Doseff, and Richard Neubig, and Kathryn Severin. We also thank Wei-Yu Liu and Yuk Kei Wan for assistance, and Christopher Buehl and Witawas Handee for discussions throughout the development of this project.

Data Sharing

Not applicable to this article as no datasets were generated or analyzed during the current study.

Funding

This work received funding from the National Institute on Aging (1R21AG051820, 1R01AG062435), the College of Osteopathic Medicine and the Office of the Vice President for Research and Graduate Studies, and the Molecular Discovery Group, Michigan State University, to M-HK, and from NIA (1R41AG057274) to DAB and M-HK.

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  1. Xiexiong Deng

    Present address: Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109-1085, USA

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Michigan State University, 603 Wilson Road, Room 401, East Lansing, MI, 48824, USA

    Mengyu Liu, Dexin Sui, Stacy Hovde, Xiexiong Deng, Kuang-Wei Wang, Hsiao-Tien Chien & Min-Hao Kuo

  2. Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI, USA

    Thomas Dexheimer

  3. Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan, China

    Hsin Lian Lin, Hsiao-Tien Chien & Yeou-Guang Tsay

  4. Department of Biological Chemistry, University of Michigan, Ann Arbor, MI, USA

    Hye Kyong Kweon, Philip C. Andrews & Roland Kwok

  5. Department of Ecology and Evolutionary Biology, University of Michigan, Ann Arbor, MI, USA

    Nora Sheen Kuo

  6. Center for Molecular Neurobiology, Department of Molecular Biochemistry, College of Medicine, The Ohio State University, Columbus, OH, USA

    Christopher A. Ayoub, Daniela Jimenez-Harrison & Jeff Kuret

  7. Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, MI, USA

    Roland Kwok

  8. Cayman Chemical, Ann Arbor, MI, USA

    Daniel A Bochar

  9. College of Veterinary Medicine, Michigan State University, East Lansing, MI, USA

    Jessica Fortin

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Contributions

ML, DS, SH, XD, and NSK constructed plasmids used in this work; ML, DS, and TD implemented p-tau aggregation assays; ML, SH, and RK developed cell-based assays for p-tau assessments; HLL, HKK, H-TC, PCA, and Y-GT performed mass spectrometry; XD, CAA, DJ-H, and JK performed transmission electronic microscopy; ML, DS, SC, and DAB produced p-tau from different PIMAX constructs; K-WW and JF performed PICUP and FTIR.

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Correspondence to Min-Hao Kuo.

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Liu, M., Sui, D., Dexheimer, T. et al. Hyperphosphorylation Renders Tau Prone to Aggregate and to Cause Cell Death. Mol Neurobiol 57, 4704–4719 (2020). https://doi.org/10.1007/s12035-020-02034-w

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