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Gain of toxic apolipoprotein E4 effects in human iPSC-derived neurons is ameliorated by a small-molecule structure corrector


Efforts to develop drugs for Alzheimer's disease (AD) have shown promise in animal studies, only to fail in human trials, suggesting a pressing need to study AD in human model systems. Using human neurons derived from induced pluripotent stem cells that expressed apolipoprotein E4 (ApoE4), a variant of the APOE gene product and the major genetic risk factor for AD, we demonstrated that ApoE4-expressing neurons had higher levels of tau phosphorylation, unrelated to their increased production of amyloid-β (Aβ) peptides, and that they displayed GABAergic neuron degeneration. ApoE4 increased Aβ production in human, but not in mouse, neurons. Converting ApoE4 to ApoE3 by gene editing rescued these phenotypes, indicating the specific effects of ApoE4. Neurons that lacked APOE behaved similarly to those expressing ApoE3, and the introduction of ApoE4 expression recapitulated the pathological phenotypes, suggesting a gain of toxic effects from ApoE4. Treatment of ApoE4-expressing neurons with a small-molecule structure corrector ameliorated the detrimental effects, thus showing that correcting the pathogenic conformation of ApoE4 is a viable therapeutic approach for ApoE4-related AD.

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We thank A. Karydas for assistance with skin biopsies, S. Ordway and T. Pak for editorial assistance and P. Davies (Albert Einstein College of Medicine) for the PHF1 antibody. This work was supported by funding from the US National Institutes of Health (grant nos. AG048030 (Y.H.), AG048017 (Y.H.), AG056305 (Y.H.) and AG023501 (B.L.M. and Y.H.)) and the California Institute for Regenerative Medicine (grant nos. RN2-00952 and TRAN1-09394; both to Y.H.) and by a gift from the Roddenberry Foundation. C.W. was partially supported by a fellowship from the California Institute for Regenerative Medicine.

Author information

C.W. and Y.H. designed and coordinated the study; C.W. performed most of the studies and data analyses; R.N. performed all MGE studies and related data analysis; Q.X. and D.J. helped with off-target analysis of gene editing and work with APOE-expressing lentiviral vectors and performed some western blots; D.W. designed and prepared the APOE-ε3 donor DNA for gene editing; D.W. and S.Y.Y. helped with genetic screening; M.E.B. helped with the cell culture; H.Y. helped with MGE studies and image collection; G.L. helped with miPSC studies; Z.A.M., B.L.M. and M.J.M. provided the APOE-null human skin biopsy; and C.W. and Y.H. wrote the manuscript.

Competing interests

Y.H. is a cofounder and scientific advisory board member of E-Scape Bio, Inc.

Correspondence to Yadong Huang.

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Supplementary Tables 1 and 2, and Supplementary Figures 1–15

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Further reading

Fig. 1: Human ApoE4/4 neurons generate more APOE fragments, have higher p-tau levels and produce more Aβ than human ApoE3/3 neurons.
Fig. 2: Increased p-tau levels in ApoE4/4 neurons are independent of higher Aβ production.
Fig. 3: ApoE4 causes GABAergic neuron degeneration and/or loss in hiPSC-derived neuronal cultures.
Fig. 4: AD-related pathologies in human ApoE4/4 neurons are specifically induced by ApoE4.
Fig. 5: ApoE4 confers a gain of toxic effects in hiPSC-derived neurons.
Fig. 6: The gain of toxic effects of ApoE4 in hiPSC-derived neurons can be ameliorated by a small-molecule structure corrector.