Investigating the molecular mechanism of P-gp/NHERF-1 network at feto maternal interface and role of paracrine signaling of EVs containing drug transporter proteins

摘要 妊娠期间的药物治疗是不可避免的，目前的干预策略并不成功， 降低妊娠相关疾病的风险。更好地了解药物转运机制 因此，需要在母胎界面（FMi）进行干预，以改善妊娠和新生儿结局。的 药物转运蛋白的表达水平随着妊娠期的进展而降低。我们的数据 表明钠氢交换调节因子-1（NHERF-1）与外排转运蛋白相互作用， 蛋白质，渗透性糖蛋白（P-gp），这种相互作用主要是在胎膜 而不是胎盘与此数据一致，外排药物机制在绒毛膜细胞中也较高， 胎膜细胞多于胎盘滋养层细胞。另一方面，来自胎儿的细胞外囊泡 膜参与旁分泌信号传导，诱导母体蜕膜的炎症变化， 子宫肌层沿着的是，EV携带药物转运蛋白作为其货物蛋白，并参与 修饰母体细胞的转运蛋白功能活性。本建议是在这些意见的基础上提出的; 核心假设是，可以靶向胎膜中的NHERF-1来调节药物 怀孕期间的交通我们将确定P-gp/NHERF-1跨胎儿的机制网络， 母体接口使用创新的FM-PLA -器官芯片（OOC）设备（Aim-1）。在目标1中，我们将使用 模拟子宫内环境的OOC，以测试P-gp底物他克莫司在FMi中的动力学。 正常的健康和疾病状态沿着有和没有NHERF-1。的表达 P-gp和NHERF-1将在整个FMi中测定。来源于胎膜的EV携带药物 转运蛋白，但它们在药物转运调节中的作用仍然未知。在Aim-2中， 探讨母体细胞对EV的摄取机制，并确定其在调节药物代谢中的作用。 使用P-gp敲除小鼠进行转运。鉴于NHERF-1和来自胎儿的EV的关键作用， 膜调节药物转运具有重要的科学和临床意义。成功 这项研究的完成将为FMi如何获得胎膜TP和EV提供新的见解 在妊娠期间的药物药代动力学中发挥作用，并将有助于 设计药物输送策略来治疗不良妊娠结局。

ABSTRACT Pharmacotherapy during pregnancy is inevitable and current interventional strategies are not successful in reducing the risks of pregnancy related disorders. A better understanding of the drug transport mechanisms involved at feto-maternal interface (FMi) is required to improve the pregnancy and neonatal outcomes. The expression levels of the drug transporter proteins decrease as the gestation period progresses. Our data suggested that sodium hydrogen exchange regulatory factor-1 (NHERF-1) interacts with efflux transporter protein, permeability glycoprotein (P-gp), and this interaction is predominantly more in the fetal membrane rather than the placenta. Consistent with this data, efflux drug mechanism is also higher in the chorion cells of fetal membrane than placenta trophoblast cells. On the other hand, extracellular vesicles derived from fetal membrane are involved in the paracrine signaling that induce inflammatory changes in maternal decidua and myometrium. Along with that, EVs carry drug transporter proteins as their cargo proteins and are involved in modifying maternal cells for transporter protein functional activities. This proposal builds on these observations; the central hypothesis is the NHERF-1 in the fetal membrane could be targeted to regulate the drug transportation during pregnancy. We will determine the mechanistic network of P-gp/NHERF-1 across fetal maternal interface using innovative FM-PLA - organ on chip (OOC) devices (Aim-1). In Aim 1, we will use the OOC that mimic an in-utero environment to test the kinetics of P-gp substrate, Tacrolimus across the FMi in normal healthy and disease conditions along with in presence and absence of NHERF-1. The expression of the P-gp and NHERF-1 will be determined across the FMi. EVs derived from the fetal membrane carry drug transporter proteins but their role in the regulation of drug transportation remains unknown. In Aim-2 we will explore the uptake mechanism of the EVs by maternal cells and determine their role in regulating drug transportation using P-gp knock down mice. Given the critical role of NHERF-1 and EVs from the fetal membrane regulates the drug transportation is of significant scientific and clinical importance. The successful completion of the study will provide novel insights into how the fetal membrane TPs and EVs derived at FMi have a role in drug pharmacokinetics during pregnancy beyond the boundaries of the placenta and will help to design drug delivery strategies to treat adverse pregnancy outcomes.