Human GATA2 adenoviral particles

Endometriosis is a gynecological disease characterized by the extrauterine growth of endometrium-like cells, leading to chronic pain and infertility. Here, researchers mapped genome-wide differences in DNA methylation between healthy human endometrium and endometriotic stromal cells and associated them with gene expression using an interaction analysis strategy. 42,248 differentially methylated CpGs were found in endometriotic and healthy cells. Significant differences in methylation were localized to 403 genes, including a disproportionate number of transcription factors. Furthermore, many of these genes have been implicated in the pathology of endometriosis and decidualization. Functional analysis of the GATA family revealed that GATA2 regulates key genes required for hormone-driven differentiation of healthy stromal cells but is hypermethylated and repressed in endometriotic cells. GATA6, which is less methylated, is abundant in endometriotic cells and effectively blocks hormone sensitivity, represses GATA2, and induces markers of endometriosis when expressed in healthy endometrial cells. A unique epigenetic fingerprint in endometriosis suggests that DNA methylation is integral to the disease and identifies a novel role for the GATA family as key regulators of uterine physiology - aberrant DNA methylation in endometriotic cells is associated with changes in GATA isoform expression, promoting progesterone resistance and disease progression.

Here, the researchers used siRNA to knock down GATA6 in OSIS while simultaneously expressing GATA2 via adenoviral (AdGATA2) transduction. xCONT scrambled siRNA and adenovirus expressing eGFP were used as controls. This was performed with or without IVD treatment for 6 days. Adenoviral-mediated expression of GATA2 increased both mRNA and protein expression of GATA2 (Figure 1A). After 6 days in culture, GATA6 siRNA reduced GATA6 mRNA by 74% and protein levels by 51%. Notably, expression of GATA2 also effectively blocked GATA6 mRNA and protein, with combined knockdown of GATA6 and overexpression of GATA2 reducing GATA6 mRNA by 95% and protein by 92% (Figure 1A). Interestingly, GATA4 mRNA expression in OSIS was reduced by GATA2 overexpression but not by GATA6 knockdown (Figure 1B). Exogenously expressed GATA2 protein was observed to be exclusively nuclear (Figure 1C).

Figure 1. The effect of silencing GATA6 and expressing GATA2 in OSIS. (Dyson M T, et al., 2014)