CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2 N141I neurons

Abstract

Basal forebrain cholinergic neurons (BFCNs) are believed to be one of the first cell types to be affected in all forms of AD, and their dysfunction is clinically correlated with impaired short-term memory formation and retrieval. We present an optimized in vitro protocol to generate human BFCNs from iPSCs, using cell lines from presenilin 2 (PSEN2) mutation carriers and controls. As expected, cell lines harboring the PSEN2 N141I mutation displayed an increase in the Aβ42/40 in iPSC-derived BFCNs. Neurons derived from PSEN2 N141I lines generated fewer maximum number of spikes in response to a square depolarizing current injection. The height of the first action potential at rheobase current injection was also significantly decreased in PSEN2 N141I BFCNs. CRISPR/Cas9 correction of the PSEN2 point mutation abolished the electrophysiological deficit, restoring both the maximal number of spikes and spike height to the levels recorded in controls. Increased Aβ42/40 was also normalized following CRISPR/Cas-mediated correction of the PSEN2 N141I mutation. The genome editing data confirms the robust consistency of mutation-related changes in Aβ42/40 ratio while also showing a PSEN2-mutation-related alteration in electrophysiology.Electronic supplementary materialThe online version of this article doi: (10.1186/s40478-017-0475-z) contains supplementary material, which is available to authorized users.

Modeling Alzheimer’s disease with iPSC-derived brain cells

“…As age is the primary risk factor for AD and other neurodegenerative disorders, it may seem counterintuitive to study AD using stem cells. It is worth noting though that even in the early stages following differentiation, neurons derived from AD patient iPSCs, and from iPSCs carrying FAD mutations, generally exhibit AD-related phenotypes such as elevated Aβ production [53][54][55]. These early alterations presumably parallel the understudied early stages of disease progression that occur in vivo.…”

“…iPSCs can be differentiated into NPCs, which can subsequently be patterned to different neuronal lineages [67][68][69]. Both passive and directed differentiation protocols having been developed for numerous different neuron subtypes, including glutamatergic, GABAergic, cholinergic and dopaminergic neurons, though existing protocols are biased strongly towards excitatory neurons [53,[68][69][70][71][72][73][74][75]. Early studies served primarily to validate the iPSC-derived models themselves and to test whether these models recapitulated findings from the large body of AD literature based on human post-mortem brain samples, rodent and other studies ( Table 2 highlights select studies of AD using iPSC-derived cells).…”

New biomarkers and therapeutic strategies in acute lymphoblastic leukemias: Recent advances

“…This technique can be used to introduce or correct disease phenotypes such as β‐thalassemia or Alzheimer disease in induced pluripotent stem cell (iPSC) …”

Gene editing in plants: assessing the variables through a simplified case study

“…CRISPR technology has provided avenues for engineering of cells, tissues, and whole organisms across the biological spectrum. Aside from the amazing potential for gene editing as a diagnostic tool and perhaps even a treatment for many debilitating diseases (Ortiz-Virumbrales et al 2017;Reczek et al 2017;Zabinyakov et al 2017), the potential possible uses of this technology in plants and livestock animals is similarly impressive (Lamas-Toranzo et al 2017). Creation of disease-resistant and less allergenic food crops (Hummel et al 2018;García-Molina et al 2019) with enhanced nutrient profiles (Wang et al 2019) is now within our reach; the future implications for feeding a progressively larger world population are immense.…”