Single‐microglia transcriptomic transition network‐based prediction and real‐world patient data validation identifies ketorolac as a repurposable drug for Alzheimer's disease

Abstract INTRODUCTION High microglial heterogeneities hinder the development of microglia‐targeted treatment for Alzheimer's disease (AD). METHODS We integrated 0.7 million single‐nuclei RNA‐sequencing transcriptomes from human brains using a variational autoencoder. We predicted AD‐relevant microglial subtype‐specific transition networks for disease‐associated microglia (DAM), tau microglia, and neuroinflammation‐like microglia (NIM). We prioritized drugs by specifically targeting microglia‐specific transition networks and validated drugs using two independent real‐world patient databases. RESULTS We identified putative AD molecular drivers (e.g., SYK , CTSB , and INPP5D ) in transition networks of DAM and NIM. Via specifically targeting NIM, we identified that usage of ketorolac was associated with reduced AD incidence in both MarketScan (hazard ratio [HR] = 0.89) and INSIGHT (HR = 0.83) Clinical Research Network databases, mechanistically supported by ketorolac‐treated transcriptomic data from AD patient induced pluripotent stem cell–derived microglia. DISCUSSION This study offers insights into the pathobiology of AD‐relevant microglial subtypes and identifies ketorolac as a potential anti‐inflammatory treatment for AD. Highlights An integrative analysis of ≈ 0.7 million single‐nuclei RNA‐sequencing transcriptomes from human brains identified Alzheimer's disease (AD)–relevant microglia subtypes. Network‐based analysis identified putative molecular drivers (e.g., SYK, CTSB, INPP5D ) of transition networks between disease‐associated microglia (DAM) and neuroinflammation‐like microglia (NIM). Via network‐based prediction and population‐based validation, we identified that usage of ketorolac (a US Food and Drug Administration–approved anti‐inflammatory medicine) was associated with reduced AD incidence in two independent patient databases. Mechanistic observation showed that ketorolac treatment downregulated the Type‐I interferon signaling in patient induced pluripotent stem cell–derived microglia, mechanistically supporting its protective effects in real‐world patient databases.