A high-throughput, quantitative platform using 2D dissociated human cerebral organoids to model neuroinflammation in Alzheimer's disease.

Neuroinflammation is a key process associated with Alzheimer's disease (AD). There is interest in developing New Approach Methodologies (NAMs) by using human in-vitro complex systems such as brain organoids, combined with machine learning and computational approaches, to reproducibly and robustly evaluate monoclonal antibodies and other therapeutic modalities on these human-derived systems. Herpesviruses such as herpes simplex virus 1 (HSV-1) had been shown to be associated with AD risk and molecular pathology. Building on top of previously reported work, we used herpes simplex virus 1 (HSV-1) infection in 2D dissociated cells from human cerebral organoids (dcOrgs) to recapitulate AD-associated molecular readouts, such as high co-abundance of intracellular beta amyloid (Aβ) and phosphorylated tau (pTau) with HSV-1. Secreted Aβ42/40 ratios in conditioned media were lower from HSV-1-infected dcOrgs, compared to mock dcOrgs. Differentially expressed transcripts from bulk and single-cell RNA sequence data in HSV-1-infected dcOrgs were enriched for AD-associated GWAS genes. Our high-throughput, quantitative framework represents a comprehensive approach to harness on the strengths of 2D dcOrgs for high-throughput applications such as therapeutic screens and can complement the 3D brain organoids and animal models for neuroinflammation in AD.