(402f) Proteoliposome Development for Placental Biomimetic Models

Currently, less than 10% of pharmaceuticals have enough information to determine their risks to a developing fetus.¹Many medications are critical and can’t be stopped during pregnancy, resulting in 9 out of 10 women in the U.S. taking medication over the course of pregnancy.² The Centers for Disease Control and Prevention (CDC) is actively raising awareness of this issue through their Treating for Two program to help identify safe treatment options to improve the health of women and babies.³ Because of this, there is a need to develop new high throughput drug screening techniques to determine molecular effects during pregnancy. Biomimetic systems representative of the placenta can aid in elucidating transport mechanisms at the maternal-fetal interface. The placenta plays an important role during pregnancy, yet it remains one of the least understood human organs.^4,5 The main cell type composing the placenta, trophoblast cells, have important functions including nutrient and waste transport, invading the endometrium to anchor the placenta, and remodeling vasculature for adequate blood flow.⁶ These placental trophoblast cells have great potential for use in developing in vitro, biomimetic models to study drug transport.

This research aims to develop placental proteoliposomes, lipid vesicles representative of the trophoblast cell membrane composition. The proteoliposomes are developed by incorporating ABCB1 (P-glycoprotein (P-gp)), a protein that is highly present in the placenta and an important drug extrusion pump, into a liposome having a lipid composition representative of placental trophoblast cells.⁷ The placental proteoliposomes are formulated using microfluidic mixing via a NanoAssemblr, which enables the incorporation of proteins into liposomes with low polydispersity.⁸ The proteoliposomes are characterized for their protein encapsulation, hydrodynamic diameter, polydispersity and zeta potential. This work aims to develop a novel platform to enable fundamental mechanistic studies of drug transport at the maternal-fetal interface. Future work will include measuring drug transport across this artificial membrane and comparing transport properties with trophoblast cell models.

References:

1. Adam MP, Polifka JE, Friedman JM. Evolving knowledge of the teratogenicity of medications in human pregnancy. Am J Med Genet. 2011;157(3):175-182. doi:10.1002/ajmg.c.30313

2. Mitchell AA, Gilboa SM, Werler MM, Kelley KE, Louik C, Hernández-Díaz S. Medication use during pregnancy, with particular focus on prescription drugs: 1976-2008. American Journal of Obstetrics and Gynecology. 2011;205(1):51.e1-51.e8. doi:10.1016/j.ajog.2011.02.029

3. Centers for Disease Control and Prevention. Treating for Two. Treating for Two. Published March 12, 2021. Accessed April 4, 2022. https://www.cdc.gov/pregnancy/meds/treatingfortwo/index.html

4. Guttmacher AE, Maddox YT, Spong CY. The Human Placenta Project: placental structure, development, and function in real time. Placenta. 2014;35(5):303-304. doi:10.1016/j.placenta.2014.02.012

5. Kaiser J. Gearing up for a closer look at the human placenta. Science. 2014;344(6188):1073. doi:10.1126/science.344.6188.1073

6. E. Davies J, Pollheimer J, Yong HEJ, et al. Epithelial-mesenchymal transition during extravillous trophoblast differentiation. Cell Adhesion and Migration. 2016;10(3):310-321. doi:10.1080/19336918.2016.1170258

7. Bailey-Hytholt CM, Shen TL, Nie B, Tripathi A, Shukla A. Placental Trophoblast-Inspired Lipid Bilayers for Cell-Free Investigation of Molecular Interactions. ACS Appl Mater Interfaces. 2020;12(28):31099-31111. doi:10.1021/acsami.0c06197

8. Molinaro R, Evangelopoulos M, Hoffman JR, et al. Design and Development of Biomimetic Nanovesicles Using a Microfluidic Approach. Adv Mater. 2018;30(15):1702749. doi:10.1002/adma.201702749