Alzheimer’s disease models in iPSC-derived glutamatergic neurons show increased secretion of pathogenic amyloid beta peptides

Veteleanu, A., et al. (2025, April) Alzheimer’s disease models in iPSC-derived glutamatergic neurons show increased secretion of pathogenic amyloid beta peptides [Poster presentation]. AD/PD25: 19th International Conference on Alzheimer’s & Parkinson’s Diseases, Vienna, Austria.

Abstract

Aims. Alzheimer’s disease (AD), a complex, multifactorial neurodegenerative disease, is challenging to study in vitro due to a lack of physiologically relevant models. ioGlutamatergic Neurons are deterministically programmed human iPSC-derived excitatory neurons that provide a consistent and scalable model to study such diseases. A panel of AD models in ioGlutamatergic Neurons was developed and characterised to determine the effects of mutations in PSEN1 and APP on amyloid beta (Aβ) production.

Methods. CRISPR/Cas9 was used to engineer heterozygous and homozygous PSEN1 M146L, APP KM670/671NL or APP V717I mutations in the parental iPSC line of the ioGlutamatergic Neurons, which were subsequently programmed using opti-ox technology to generate the disease model cells. The disease models were cultured for 30 days alongside their genetically matched wild type control. Supernatant was collected on days 10, 20 and 30, and concentrations of Aβ38, Aβ40 and Aβ42 were determined using the V-PLEX A-beta Peptide Panel ELISA kit.

Results. ioGlutamatergic Neurons carrying the PSEN1 M146L and APP V717I mutations secreted significantly more Aβ42 compared to their wild type control, showing higher Aβ42:40 ratios at days 20 and 30. Importantly, a clear correlation between genotype and Aβ42:40 ratios was observed, as wild type, heterozygous and homozygous mutants showed a stepwise increase in Aβ42 production relative to Aβ40. ioGlutamatergic Neurons APP KM670/671NL secreted significantly more Aβ38, Aβ40, and Aβ42 than their wild type control, but the Aβ42:40 ratio did not increase, as expected.

Conclusions. ioGlutamatergic Neurons with mutations in PSEN1 or APP recapitulate the increase in Aβ42 secretion observed in Alzheimer’s patients. This demonstrates their validity as an in vitro model to study AD and for the discovery of drugs targeting the pathogenic Aβ pathway.