Dolutegravir Developmental Toxicity is Mitigated by Magnesium and Folate in Zebrafish Embryos

Abstract

Integrase-strand-transfer inhibitors have transformed HIV therapy, yet the widely prescribed drug dolutegravir (DTG) has been linked to developmental toxicity and its teratogenic mechanism remains uncertain. Here we use zebrafish to dissect DTG toxicity during early vertebrate development. DTG exposure from 2-4 h post-fertilization (hpf) to 24 hpf produced high mortality and abnormal morphology; co-treatment with folic acid or 5-methyltetrahydrofolate partially restored normal morphology, whereas calcium had no effect. Strikingly, supplementation with magnesium (Mg) rescued DTG-exposed embryos almost as effectively as folate, implicating magnesium availability in protection. In competitive binding assays, Mg increased binding of folate to purified folate receptor (FOLR1) by 30% in the presence of DTG. Maternal-zygotic zebrafish folr1 mutant embryos contained significantly less endogenous folate than wild-type embryos yet displayed marked hypersensitivity to DTG that could not be mitigated by folate supplementation. Critically, magnesium supplementation partially rescued DTG toxicity in folr1 mutants, indicating a Folr1-independent component and placing the free DTG vs Mg-bound DTG balance upstream of folate transport. These results support a model in which free DTG antagonizes FOLR1, and Mg modifies DTG developmental toxicity by both FOLR1 dependent and independent processes. Our work identifies magnesium status as a modifiable determinant of DTG teratogenicity and provides a proof-of-concept zebrafish model that could be adapted for rapid in vivo screening of integrase inhibitors for developmental phenotypes.