May 19 2021 1:30 pm
High throughput screening for identification of compounds that induce fusion in BeWo cells
Dr. Amir Faisal
Zoom Meetings (Online)
MS Thesis defense
The placenta is the first organ that forms during mammalian development for the in utero development of the fetus. Placenta formation starts as the blastocyst adheres to the maternal endometrium. The cytotrophoblast cells present in the outer trophectoderm layer of blastocyst undergo cell fusion to give rise to morphologically differentiated, invasive, and non-proliferative syncytiotrophoblast cells. The syncytiotrophoblast cells invade the maternal decidua by degrading the endometrial stromal tissues. Defective invasion of cytotrophoblast cells through endometrium can lead to various pregnancy-related complications, including preeclampsia, restricted fetal growth, and abortions. The molecular basis of differentiation, invasion and migration of cytotrophoblast cells through the maternal endometrium is not well understood. The human choriocarcinoma BeWo cell line has been widely used as a model to study cytotrophoblast fusion and differentiation in vitro. In this study, we aimed to identify novel regulators of BeWo cell fusion and differentiation through a chemical genetic screen. A library of 512 inhibitors was screened in BeWo cells for the identification of novel regulators of BeWo cell fusion. Cells were treated with 1µM inhibitors for 72 hours in 96-well plates and stained with DAPI followed by imaging through confocal microscopy. Images were analyzed, and fold change in nuclear size was determined. Out of the total 1127 inhibitors screened previously and, in this study, 83 inhibitors 2 showed a fold increase in nuclear size greater than the positive control. The hits identified through primary screening will be validated through secondary screening. The target proteins identified through the screening will not only help understand the pathways involved in syncytialization and polyploidy in BeWo cells but can also act as potential therapeutic targets for drug development to treat placental abnormalities.