Abstract: Induced overexpression of Neurogenin 2 (iNGN2) is a valuable strategy to accelerate the conversion of human induced pluripotent stem cells (hiPSCs) into neurons for modeling of neurodegenerative diseases such as Alzheimer’s Disease (AD). However, iNGN2 alone generates neurons of a mixed regional identity, potentially limiting their relevance for modeling AD. We developed an alternative neural induction strategy, which prepatterns hiPSCs before NGN2 induction to obtain a stronger cortical identity. This strategy involves inhibiting different pathways critical for self-renewal and meso-endodermal differentiation, independent of commonly applied dual SMAD inhibition (dSMADi). Combined with iNGN2, this induction paradigm quickly leads to homogeneous cultures of excitatory cortical neurons. Furthermore, the cells are responsive to regional patterning cues during the short neural induction pulse. This enables generation of neurons from different regions of both the central and peripheral neuron systems, including midbrain dopaminergic, motoneurons and sensory neurons, improving in vitro models for a range of neurological diseases and neurodegenerative disorders. Cortical neurons generated with this paradigm are suitable for an AD-relevant tau aggregation assay, similar in performance to cortical neurons derived from the dSMADi protocol, highlighting their applicability in disease – relevant in vitro models.
Disclaimer: CH, AW, MJH, NN, CS, LR, TL, CU, LNM, JK, DG, DS, HL, LB, BMS, MB, RW, JR, IW, JDM, MC, PR are employees of AbbVie. VH was employee of AbbVie at the time of the study. AK was employee of the Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences at the time of the study and was funded by the Graduiertenkolleg TASCDT of the MWK Baden-Württemberg and the Albert und Annelise Konanz-Stiftung. R.R and M.H. are current employees of Center for Mass Spectrometry and Optical Spectroscopy, Mannheim University of Applied Sciences and Institute of Medical Technology, Heidelberg University and Mannheim University of Applied Sciences and have no funding to disclose. The design, study conduct, and financial support for this research were provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication.
