Prenatal caloric restriction reprograms endothelial transcriptional states and blood-brain barrier integrity: Implications for neurodevelopmental disorders

Prenatal caloric restriction (RP) induces long-lasting physiological adaptations that shape adult metabolism and brain function, yet its impact on cerebrovascular programming and responses to later-life nutritional stress remains poorly understood. We previously showed that RP offspring exhibit hypoactivity and anxiety-like behaviors in adulthood, suggesting persistent neurobehavioral alterations. Here, we investigated how RP acts as a developmental "first hit" to program brain endothelial transcriptional states, and how a subsequent adult caloric restriction (CR) functions as a "second hit" that adjust these pre-established programs in mice. Using RNA sequencing combined with cell-type deconvolution, we profiled endothelial transcriptional states across four nutritional conditions: ad libitum controls (AL-AL), adult restriction only (AL-CR), prenatal restriction only (RP-AL), and combined restriction (RP-CR). RP induced robust transcriptional changes, consistent with endothelial reprogramming characterized by enrichment of angiogenic tip-cell signatures and modulation of genes involved in blood-brain barrier integrity and vascular stability. Importantly, these prenatal imprints conditioned the endothelial transcriptional response to adult CR, revealing altered adaptive plasticity rather than a naïve angiogenic response. Notably, three genes-Kdr, Hdac7, and Mmrn2-were significantly regulated and strongly correlated with GSVA scores of angiogenic pathways, particularly under the two-hit dietary paradigm. Collectively, these genes integrate pro-angiogenic signaling (Kdr), epigenetic control of endothelial identity (Hdac7), and extracellular matrix-mediated vascular stabilization (Mmrn2), collectively defining endothelial states shaped by recurrent nutritional stress. Our findings support a two-hit model in which early-life transcriptional reprogramming compromises neurovascular integrity, potentially establishing a vascular vulnerability that contributes to the etiology of psychiatric disorders such as schizophrenia.