In-vitro gata2 overexpression reverses decidualization defects by improving endometrial receptivity markers in human endometrial stromal cells from patients with adenomyosis

GATA binding protein 2 and 6 (GATA2 and GATA6) are zinc-finger transcription factors involved in proliferation of hematopoietic and endocrine cell lineages, as well as cell differentiation and organogenesis, respectively. Gata2 deficient mice are infertile due to impaired uterine response associated with lower PGR levels and attenuated progesterone signaling resulting in defective decidualization (1,2). Elevated GATA6 increases aromatase levels with concomitant aberrant activation of estrogen synthesis (3). We previously reported that several decidualization mediators such as LIF, IL11R and PGR are dysregulated in endometrium of adenomyosis patients during the window of implantation (4). Our previous immunohistochemistry data revealed an overall 4-fold lower GATA2 and 4-fold higher GATA6 H-SCORE level in endometrial stromal cells of adenomyosis patients vs. controls (5). Thus, we hypothesized that dysregulated GATA2 and GATA6 expression contribute to adenomyosis-associated implantation failure by impairing decidualization. Human endometrial stromal cells (HESCs) were isolated from patients with confirmed adenomyosis and from fertility proven leiomyoma patients as controls. Control and adenomyosis HESC cultures (n=4/each) were treated with either placebo or 10-8 M estradiol (E2), or decidualization media (EMC) containing 10-8M E2, 10-7M medroxyprogesterone acetate, and 5x10-5M cAMP for 6 and 10 days. GATA2, insulin like growth factor binding protein 1 (IGFBP1), prolactin (PRL), PGR, estrogen receptor-α/β (ESR1/2), LIF, and ILR11 mRNA levels were analyzed by qPCR. Additionally, control HESC cultures (n=4) were transfected with scrambled siRNA (control) or GATA2-specific siRNAs for 6 days while adenomyosis HESC cultures (n=4) were transfected with GATA2-specific expression vector to induce GATA2 overexpression, and corresponding mRNA levels of above factors were analyzed after confirming GATA2 silencing or overexpression by qPCR and immunoblotting. Decidual induction with EMC resulted in significantly lower IGFBP1, LIF and IL11R, but not ESR1 or ESR2 mRNA levels in HESC cultures from adenomyosis patients vs. controls (P<0.05). GATA2 silencing in control HESC resulted in an adenomyosis-like state with significant reductions in GATA2 levels (P<0.001) and accompanying dysregulation of PGR, PRL, ESR1, LIF and IL11R levels. Finally, GATA2 overexpression in adenomyosis HESCs caused restoration of the defective decidual response with significant increases in GATA2, PGR, PRL and LIF mRNA levels (P<0.05). In-vitro results demonstrate that GATA2 silencing in HESCs obtained from normal (fertile) women impairs important decidualization and implantation factors such as ESR1, PGR, IGFBP1, PRL, LIF and IL11R, whereas GATA2 overexpression improves regulation of these markers in HESCs obtained from adenomyosis patients. These results suggest that GATA2 inducing factors have therapeutic potential for adenomyosis.