Profiling RNA Cargo in Extracellular Vesicles from hiPSC-Derived Neurons of Alzheimer's Disease Patients
Alzheimer's disease (AD) is a major neurodegenerative disorder that affects more than 55 million people, with an incidence that is projected to triple by 2050. Despite continuous advancements in the field, reliable treatment and early detection strategies remain elusive. Extracellular vesicles (EVs) play a major role in cellular communication throughout the body. In this study, we assessed the cargo of neuronal-specific EVs for their potential as AD biomarkers. We isolated EVs released from iPSC-derived excitatory glutamatergic neurons generated from eight AD patients (ADiNEVs) and six healthy controls (iNEVs). We performed RNA-sequencing and identified significant differences in RNA cargo between ADiNEVs and iNEVs. Notably, fewer small nuclear RNAs (snRNAs) were found in ADiNEVs. RNA transcripts significantly more abundant in ADiNEVs included MT-CO1, PRR32, and IGSF8 messenger RNAs. We also observed fewer XIST long noncoding RNAs and miR-7-5p microRNA content in ADiNEVs. These findings suggest that precision medicine approaches, such as characterizing the content of EVs from a patient's own cells, could advance early detection and management of AD. Overall design: We generated iPSC-derived excitatory glutamatergic neurons from 14 human participants, including 8 individuals diagnosed with sporadic Alzheimer's disease (sAD) and 6 age-matched healthy controls. Then we collected EVs from neuronal culture supernatants on Day 21 of differentiation, extracted RNAs from EVs, and performed small RNA sequencing on these EVs using the NovaSeq 6000 platform and S4 Reagent Kit v1.5.