Development and utilization of human decidualization reporter cell line uncovers new modulators of female fertility

Abstract

Failure of embryo implantation accounts for a significant percentage of female infertility. Exquisitely coordinated molecular programs govern the interaction between the competent blastocyst and the receptive uterus. Decidualization, the rapid proliferation and differentiation of endometrial stromal cells into decidual cells, is required for implantation. Decidualization defects can cause poor placentation, intrauterine growth restriction, and early parturition leading to preterm birth. Decidualization has not yet been systematically studied at the genetic level due to the lack of a suitable high-throughput screening tool. Herein we describe the generation of an immortalized human endometrial stromal cell line that uses yellow fluorescent protein under the control of the prolactin promoter as a quantifiable visual readout of the decidualization response (hESC-PRLY cells). Using this cell line, we performed a genome-wide siRNA library screen, as well as a screen of 910 small molecules, to identify more than 4,000 previously unrecognized genetic and chemical modulators of decidualization. Ontology analysis revealed several groups of decidualization modulators, including many previously unappreciated transcription factors, sensory receptors, growth factors, and kinases. Expression studies of hits revealed that the majority of decidualization modulators are acutely sensitive to ovarian hormone exposure. Gradient treatment of exogenous factors was used to identify EC50 values of small-molecule hits, as well as verify several growth factor hits identified by the siRNA screen. The high-throughput decidualization reporter cell line and the findings described herein will aid in the development of patient-specific treatments for decidualization-based recurrent pregnancy loss, subfertility, and infertility.

Capture and metabolomic analysis of the human endometrial epithelial organoid secretome

“…Hormonal supplementation is lacking from the current experimental design as existing data indicate that the stage of the menstrual cycle does not impact UF carbohydrate (39) or amino acid (43) composition, barring possibly taurine (83). However, investigating whether EOF composition is hormonally responsive and functionally relevant, e.g., capable of inducing stromal cell decidualization in vitro (84,85), represents an interesting line for pursual. Furthermore, probing the influence of a pathophysiological endocrine milieu-mimicking aspects of subfertilityassociated disorders such as polycystic ovary syndrome (i.e., hyperandrogenism) and diabetes mellitus (i.e., hyperglycemia)-on the EEO IOF and EOF secretome, may shed light on several metabolomic etiologies of pregnancy loss.…”

Signaling through retinoic acid receptors is essential for mammalian uterine receptivity and decidualization

“…In our previous study, we engineered a fluorescent reporter hESC line and performed genome-wide siRNA screening to identify genes required for normal decidualization ( 44 ). A total of 136 genes involved in the RA pathway were among the hits, including 29 upstream and 107 downstream of RAR signaling ( Supplemental Figure 4 ).…”

“All-In-One” Genetic Tool Assessing Endometrial Receptivity for Personalized Screening of Female Sex Steroid Hormones

“…Interestingly, almost simultaneously with our research, another group developed a high-throughput screening tool to study the molecular regulators of decidualization at the genetic level (Haller et al, 2019 ). Specifically, the authors used immortalized human ESC line transduced with the genetic construct that upon recombination carried yellow fluorescent protein under the PRL promoter.…”