Beyond the amyloid hypothesis: leveraging human-centered complex in vitro models to decode Alzheimer’s disease etiology

Alzheimer's disease (AD) is a complex neurodegenerative condition and the leading cause of dementia worldwide. Treatments that safely and effectively counteract disease progression are currently lacking. While the formation of amyloid plaques has long been considered the leading hypothesis of disease onset, growing evidence suggests that the emergence of AD could be driven by a combination of underlying factors that promote chronic neuroinflammation, including pathogenic infections, environmental toxicants, and disruptions along the gut-brain axis. Traditional nonclinical models of AD, such as monolayer cell cultures and transgenic mice, struggle to capture the complexity of the disease as it occurs in humans. Human-centered complex <i>in vitro</i> models (CIVMs), including cerebral organoids and microfluidic organ-on-a-chip (OOC) technologies, provide greater physiological relevance by more closely recapitulating key cellular and molecular features of the human brain and disease mechanisms. In this mini review, we evaluate recent advances in CIVMs and how they are being leveraged to investigate emerging hypotheses of AD etiology. Cerebral organoids and OOC platforms can consistently replicate neuropathological hallmarks of neurodegeneration in response to pathogenic or environmental insults, including blood-brain barrier disruption, amyloid-β accumulation, tau hyperphosphorylation, and glial activation. We also highlight early efforts to model the gut-brain axis using organoid and multi-OOC systems, demonstrating how microbiota-derived factors can affect neural processes. Collectively, these studies show that human-centered CIVMs can be applied to both recreate and mechanistically disentangle interrelated pathological processes to an extent beyond that afforded by animal models, thus offering new opportunities to identify causal mechanisms and potential therapeutic targets.