Dysregulated Expression of GATA2 and GATA6 Transcription Factors in Adenomyosis: Implications for Impaired Endometrial Receptivity.

Abstract

OBJECTIVE:

To study the effect of adenomyosis on the localized expression of the GATA binding protein 2 and 6 (GATA2 and GATA6) zinc-finger transcription factors that are involved in proliferation of hematopoietic and endocrine cell lineages, and cell differentiation and organogenesis, potentially leading to impaired endometrial implantation.

DESIGN:

Laboratory based experimental study.

SETTING:

Academic Hospital/Laboratory.

PATIENTS:

Human endometrial stromal cells (HESCs) of reproductive age patients, 18-45 years of age, with adenomyosis compared to patients with no pathology/leiomyomatous uteri as controls (n=4 in each group respectively). Additionally, mid-secretory phase endometrial sections were obtained from adenomyosis patients and leiomyoma control patients (n=8 in each group, respectively).

INTERVENTIONS:

GATA2 and GATA6 immunohistochemistry (IHC) and H-SCORE were performed on the mid-secretory phase endometrial sections from adenomyosis and leiomyoma control patients (n=8 each, respectively). Control and adenomyosis patient HESC cultures were treated with 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. 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 human GATA2 expression vectors to silence or induce GATA2 overexpression.

MAIN OUTCOME MEASURES:

Immunohistochemistry was performed to obtain GATA2 and GATA6 H-SCORES in adenomyosis vs. control patient endometrial tissue. Expression of GATA2, GATA6, Insulin like growth factor binding protein 1 (IGFBP1), Prolactin (PRL), Progesterone receptor (PGR), Estrogen receptor 1 (ESR1), leukemia inhibitory factor (LIF), and Interleukin receptor 11 (IL11R) mRNA levels were analyzed by qPCR with normalization to ACTB. Silencing and overexpression experiments also had the corresponding mRNA levels of the above factors analyzed. Western blot analysis was performed on isolated proteins from transfection experiments.

RESULTS:

IHC revealed an overall 4-fold lower GATA2 and 4-fold higher GATA6 H-SCORE level in the endometrial stromal cells of adenomyosis patients vs. controls (P<0.01). Decidual induction with EMC resulted in significantly lower GATA2, PGR, PRL and IGFBP1 mRNA levels in HESC cultures from adenomyosis patients vs. controls (P<0.05). LIF and IL11R mRNA levels were also significantly dysregulated in adenomyosis HESCs compared to controls (P<0.05). Silencing of GATA2 expression in control HESCs induced an adenomyosis-like state with significant reductions in GATA2 (P<0.001), increases in GATA6 (P<0.01) and accompanying aberrations in PGR, PRL, ESR1 and LIF levels. Conversely, GATA2 overexpression via vector in adenomyosis HESCs caused partial restoration of the defective decidual response with significant increases in GATA2, PGR, PRL and LIF expression (P<0.01-0.05).

CONCLUSION:

In-vivo and in-vitro results demonstrate that there is an overall inverse relationship between endometrial GATA2 and GATA6 levels with adenomyosis patients having diminished GATA2 and concurrent elevated GATA6 levels. Additionally, lower GATA2 and higher GATA6 levels together with aberrant levels of important receptors and implantation factors such as ESR1, PGR, IGFBP1, PRL, LIF and IL11R mRNA in HESCs from adenomyosis patients or GATA2-silenced control HESCs, supports impaired decidualization. These effects were partially restored with GATA2 overexpression in adenomyosis HESCs demonstrating a potential therapeutic target.

• Authors: J Pavlovic Zoran, H Pai Angel, Hsiao Tzu-Ti, Yen Chih-Feng, Alhasan Hasan, Ozmen Asli, P New Erika, Guo Xiaofang, N Imudia Anthony, Guzeloglu-Kayisli Ozlem, J Lockwood Charles, A Kayisli Umit
• Journal: F&S science (F S Sci) (Nov 14 2023) ISSN: 2666-335X [Electronic] United States
• PMID: 37972693 (Publication Type: Journal Article)
• Copyright: Copyright © 2023. Published by Elsevier Inc.