Maternal, placental and fetal response to a non-viral, polymeric nanoparticle gene therapy in nonhuman primates

Abstract

BackgroundCurrently, there are no placenta-targeted treatments to alter thein uteroenvironment. Water-soluble polymers have a distinguished record of clinical relevance outside of pregnancy. We have demonstrated the effective delivery of polymer-based nanoparticles containing a non-viral humaninsulin-like 1 growth factor(IGF1) transgene to correct placental insufficiency in small animal models of fetal growth restriction (FGR). Our goal was to extend these studies to the pregnant nonhuman primate (NHP) and assess maternal, placental and fetal responses to nanoparticle-mediatedIGF1treatment.MethodsPregnant macaques underwent ultrasound-guided intraplacental injections of nanoparticles (GFP-orIGF1-expressing plasmid under the control of the trophoblast-specificPLAC1promoter complexed with a HPMA-DMEAMA co-polymer) at approximately gestational day 100 (term = 165 days). Fetectomy was performed 24 h (GFP; n =1), 48 h (IGF1; n = 3) or 10 days (IGF1; n = 3) after nanoparticle delivery. Routine pathological assessment was performed on biopsied maternal tissues, and placental and fetal tissues. Maternal blood was analyzed for complete blood count (CBC), immunomodulatory proteins and growth factors, progesterone (P4) and estradiol (E2). Placental ERK/AKT/mTOR signaling was assessed using western blot and qPCR.FindingsFluorescent microscopy and in situ hybridization confirmed placental uptake and transgene expression in villous syncytiotrophoblast. No off-target expression was observed in maternal and fetal tissues. Histopathological assessment of the placenta recorded observations not necessarily related to theIGF1nanoparticle treatment. In maternal blood, CBCs, P4 and E2 remained within the normal range for pregnant macaques across the treatment period. Changes to placental ERK and AKT signaling at 48 h and 10 d afterIGF1nanoparticle treatment indicated an upregulation in placental homeostatic mechanisms to prevent over activity in the normal pregnancy environment.InterpretationMaternal toxicity profile analysis and lack of adverse reaction to nanoparticle-mediatedIGF1treatment, combined with changes in placental signaling to maintain homeostasis indicates no deleterious impact of treatment.FundingNational Institutes of Health, and Wisconsin National Primate Research Center.