RNA-seq Analysis of Changes in mRNA Expression for the hTERT-HMA/B Human Myometrial Cell Line treated with all Combinations of Progesterone, Forskolin, and Interleukin-1ß

Parturition involves cellular signaling changes driven by the complex interplay between progesterone (P4), inflammation, and the cyclic adenosine monophosphate (cAMP) pathway. To characterize this interplay, comprehensive transcriptomic studies utilizing 8 treatment combinations were performed on the hTERT-HMA/B telomerase immortalized human myometrial cell line and myometrium obtained from term c-section deliveries. Genome-wide RNA-sequencing was performed on total RNA extracted from hTERT-HMA/B cells treated with all combinations of P4, forskolin (induces cAMP), and interleukin-1ß (IL-1ß). Gene set enrichment and regulatory network analyses was then used to identify pathways commonly, differentially, or synergistically regulated by each treatment. Tissue similarity index (TSI) was also applied to characterize the correspondence between cell lines and tissue phenotypes. In addition to their individual anti-inflammatory effects, P4 and cAMP synergistically blocked specific inflammatory pathways/regulators including STAT3/6, CEBPA/B, and OCT1/7, but not NFKB. TSI analysis indicated that forskolin+P4- and IL-1ß-treated cells exhibit transcriptional signatures highly similar to non-laboring and laboring term myometrium, respectively. The data identify potential therapeutic targets to prevent preterm birth and show that the hTERT-HMA/B cell line provides is an accurate transcriptional model for term ± labor myometrium. Overall design: Comparisons made between all single stimulus treated cells (P4, FSK, IL-1ß) with untreated cells to identify signaling by progesterone, cAMP, and IL-1ß and comparisons made between IL-1ß conditions to identify anti-inflammatory actions of P4 and/or FSK in the presence of IL-1ß.