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O1‐06‐01: Modeling tauopathies in human pluripotent stem cells
Author(s) -
Karch Celeste,
Martinez Rita,
Hsu Simon,
Ezerskiy Lubov,
Norton Joanne,
Goate Alison
Publication year - 2015
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2015.07.057
Subject(s) - induced pluripotent stem cell , tau protein , biology , tau pathology , microbiology and biotechnology , mutation , neuroscience , gene , genetics , embryonic stem cell , alzheimer's disease , disease , pathology , medicine
Background: Tauopathies are a class of neurodegenerative diseases that are characterized by hyperphosphorylated tau aggregates in the brain. In a subset of tauopathies, genetic changes in MAPT, the gene encoding the tau protein, are sufficient to initiate a cascade of events that leads to tau aggregation and death of neuronal populations in the brain. Increasing evidence suggests that tau aggregates spread along neuronal networks in the brain via tau release and tau uptake. However, the cascade of events that leads to disease remains poorly understood.Methods:We sought to develop a stem cell model of tauopathies that captures the genetic complexities of the MAPT gene and the phenotypic complexities of the human neuron. We generated human induced pluripotent stem cell (iPSC) using nonintegrating Sendai virus from dermal fibroblasts carryingMAPTmutations (P301L, IVS10+16, V337M, R406W). We measured tau phosphorylation, release and aggregation in iPSC-derived neurons from MAPT mutation carriers and controls. To determine if correction of MAPT mutations restores normal tau metabolism, we developed a pipeline for efficient, seamless modification of point mutations using CRISPR/Cas9 technology. Applying this pipeline to a MAPT P301L iPSC line, we generated isogenic, iPSC clones that are corrected toWTor that are modified toMAPTP301S.Results: Consistent with our findings in immortalized cell models, we found that tau is actively released from human iPSC-derived neurons via the unconventional secretory pathway and that calcium signaling plays an important role in tau release. We found that the rate of tau release was modified in iPSC-derived neurons fromMAPTmutation carriers compared with controls. Reversion of the MAPT P301L to WT using CRISPR/Cas9 restored the tau release rate to the rates observed in unrelated control lines. Additionally, human iPSCderived neurons endogenously expressingMAPTmutations produced multimeric tau species that were absent in unrelated and isogenic control lines. Conclusions:Together, these stem cell models capture key pathological hallmarks of Alzheimer’s disease and other tauopathies and, thus, are beginning to define the cascade of events that lead to neurodegenerative tauopathies, providing avenues for advancement in therapeutic intervention.