HAND1 knockdown disrupts trophoblast gene expression in vitro in Bewo cells

Congenital heart disease (CHD) affects nearly 1% of births annually, and neonatal and perinatal outcomes in cases of CHD are strongly impacted by pregnancy outcomes. CHD pregnancies carry increased risk of developing pathologies of abnormal placentation including fetal growth restriction, preeclampsia, preterm birth, and stillbirth. HAND1 is a gene associated with CHD. In this study, we aimed characterize the mechanistic impacts of disrupting HAND1 on human placenta trophoblast cell lines. Overall design: To silence HAND1 gene expression, BeWo cells, a choriocarcinoma model of human cytotrophoblasts, were treated for 96 h with HAND1/2 siRNA. Cells were harvested and mRNA isolated. RNA sequencing was performed using the Illumina NextSeq 550 platform (single end reads 85-101bp, read depth 25M). The Galaxy Bioinformatic Platform was used for quality control, alignment and generation of read counts. Normalization and differential gene expression analyses were conducted using general linear modeling in edgeR packages. Statistical significance was determined using a log2 fold change cut off >1.0 or <-1.0 and unadjusted =0.05. Overrepresentation analysis was performed using Panther DB for Reactome pathways and GO Biological Processes, and String DB for protein association network analysis.