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

研究胎儿母体界面P-gp/NHERF-1网络的分子机制以及含有药物转运蛋白的EV的旁分泌信号传导的作用

• 批准号: 10748250
• 负责人: Ananth kumar Kammala
• 金额: $ 35.64万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10748250
• 关键词: ABCG2 gene; Binding; Binding Proteins; Biological Assay; Bispecific Antibody 2B1; Carrier Proteins; Categories; Cell surface; Cells; Chorion; Clinical; Compensation; Complement; DLG4 gene; Data; Decidua; Decidua Basalis; Decidual Cell; Devices; Disease; Down-Regulation; Drug Delivery Systems; Drug Design; Drug Efflux; Drug Kinetics; Drug Regulations; Drug Transport; Drug resistance; Dyes; Environment; Evaluation; Exposure to; Family; Fetal Membranes; Fetus; Gene Silencing; Glycoproteins; Human; Hydrogen; Image; Individual; Inflammation; Inflammatory; Inflammatory Response; Intervention; Kinetics; Knockout Mice; Knowledge; Label; Lipopolysaccharides; Maternal-Fetal Exchange; Microscopy; Modeling; Molecular; Mothers; Mus; Nutrient; OATP Transporters; Organ; Organ Model; Oxidative Stress; Paracrine Communication; Pathologic; Pathway interactions; Patients; Permeability; Pharmaceutical Preparations; Pharmacotherapy; Placenta; Pregnancy; Pregnancy Outcome; Proteins; Regulation; Risk; Risk Factors; Role; Signal Induction; Small Interfering RNA; Sodium; Stains; Tacrolimus; Technology; Teratogens; Testing; Time; Tissues; Transportation; Western Blotting; adverse pregnancy outcome; cigarette smoke; cytotrophoblast; decidua parietalis; drug mechanism; extracellular vesicles; fetal; fetus cell; improved; in utero; in vitro Model; in vivo Model; inhibitor; innovation; insight; knock-down; lipophilicity; live cell imaging; microphysiology system; myometrium; neonatal outcome; novel; organ on a chip; particle; pregnant; protein function; response; sodium-hydrogen exchanger regulatory factor; solute; transcriptome; trophoblast; uptake

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.

抽象的 妊娠期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 改善妊娠和新生儿结局需要参与胎儿-母体界面（FMi）。这 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)，这种相互作用主要发生在胎膜中 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。 胎膜滋养层细胞高于胎盘滋养层细胞。另一方面，源自胎儿的细胞外囊泡 膜参与旁分泌信号传导，诱导母体蜕膜和炎症变化 子宫肌层。除此之外，电动汽车携带药物转运蛋白作为其货物蛋白，并参与 修改母体细胞的转运蛋白功能活动。该提案建立在这些观察的基础上； 中心假设是胎膜中的 NHERF-1 可以靶向调节药物 怀孕期间的交通。我们将确定 P-gp/NHERF-1 在胎儿中的机制网络 母体接口使用创新的 FM-PLA - 片上器官 (OOC) 设备 (Aim-1)。在目标 1 中，我们将使用 OOC 模拟子宫内环境，测试 P-gp 底物、他克莫司穿过 FMi 的动力学 正常健康和疾病状况以及是否存在 NHERF-1。的表达式为 P-gp 和 NHERF-1 将通过 FMi 确定。源自胎膜的 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。在目标 2 中，我们将 探索母体细胞对EV的摄取机制并确定其在药物调节中的作用 使用 P-gp 敲低小鼠进行运输。鉴于 NHERF-1 和来自胎儿的 EV 的关键作用 膜调节药物运输具有重要的科学和临床意义。成功者 该研究的完成将为胎膜 TP 和 EV 如何在 FMi 中衍生提供新的见解 在妊娠期间超越胎盘边界的药物药代动力学中发挥作用，并有助于 设计药物输送策略来治疗不良妊娠结局。