This preprint reports that prenatal alcohol exposure (PAE) downregulates focal adhesion molecules Tensin 1 (Tns1) and Tensin 2 (Tns2) in brain microvascular endothelial cells (BMVECs) and that these changes contribute to impaired sprouting angiogenesis. The microRNA miR-150-5p, which previous work showed is elevated by PAE, was found to directly regulate Tns1 and Tns2 via binding to their 3′ untranslated regions. Overexpressing Tns1 and Tns2 restored endothelial sprouting that was suppressed by ethanol and by miR-150-5p modulation, linking molecular regulation to a functional angiogenic readout.
Fetal alcohol spectrum disorders (FASD) arise from prenatal alcohol exposure and produce developmental defects during embryogenesis. Among affected developmental programs, angiogenesis in the developing brain is particularly vulnerable but remains incompletely characterized. Identifying molecules and regulatory pathways altered by PAE that impair cerebrovascular development could clarify mechanisms of FASD-related brain dysfunction.
Using brain microvascular endothelial cells as a model for cerebrovascular endothelium, the authors observed significant reductions in expression levels of the focal adhesion proteins Tns1 and Tns2 following alcohol exposure. Tensins are focal adhesion-associated scaffolding proteins implicated in cell–matrix interactions and cytoskeletal dynamics; reductions in these proteins provide a plausible mechanistic link between PAE and endothelial dysfunction.
The study connects the observed decrease in Tensins to microRNA-mediated regulation. The microRNA miR-150-5p, previously reported by the authors to be increased after PAE, was shown here to regulate Tns1 and Tns2 expression. Dual-luciferase reporter assays demonstrated direct binding of miR-150-5p to the 3′UTRs of both Tns1 and Tns2, supporting a post-transcriptional repression mechanism. These results position miR-150-5p as a mediator linking ethanol exposure to decreased focal adhesion protein expression in BMVECs.
To evaluate the functional consequences of altered Tns1/Tns2 and miR-150-5p levels, the authors used sprouting angiogenesis bead assays, a three-dimensional endothelial sprouting model. Ethanol treatment reduced endothelial sprouting in this assay. Overexpression of Tns1 and/or Tns2 rescued the sprouting deficits induced by ethanol as well as those produced by miR-150-5p modulation. These rescue experiments indicate that restoring Tensin expression can counteract both ethanol- and miRNA-mediated impairments in endothelial sprouting.
The data support a model in which PAE elevates miR-150-5p, leading to downregulation of Tns1 and Tns2, disruption of focal adhesion pathways in BMVECs, and consequent impairment of sprouting angiogenesis. Restoring Tns1/Tns2 expression reversed angiogenic deficits in vitro, suggesting that targeting this pathway could be a strategy to ameliorate cerebrovascular dysfunction resulting from prenatal alcohol exposure. The study provides mechanistic insight into how molecular changes induced by alcohol exposure can translate into altered vascular development.
Key experimental approaches reported in the abstract include measurement of Tns1 and Tns2 expression in BMVECs after alcohol exposure, miRNA regulation studies linking miR-150-5p to Tns1/Tns2, dual-luciferase assays to test direct 3′UTR binding, and sprouting angiogenesis bead assays to quantify endothelial sprouting and assess rescue by Tensin overexpression. The work was funded by the National Institutes of Health (R01AA031486). The authors declared no competing interests. The report is posted as a bioRxiv preprint.
The abstract describes core molecular and functional findings but does not provide full experimental detail, quantitative effect sizes, in vivo validation, or long-term developmental outcomes. Those details are not reported in the abstract and would require consulting the full preprint for methods, statistical analyses, and expanded results. Future work could clarify dose-response relationships, in vivo relevance during embryogenesis, and whether targeting the miR-150-5p–Tensin axis modifies behavioral or cerebrovascular endpoints in animal models of PAE.
This research is reported as a bioRxiv preprint (posted October 7, 2026) by Richard J. Justice and Amy S. Gardiner, with DOI https://doi.org/10.64898/2026.10.06.757079. The preprint is available under a CC-BY-NC 4.0 license. The authors declared no competing interests.