Zoran Jason Pavlovic M.D. , Angel Hsin-Yu Pai M.D. , Tzu-Ti Hsiao M.S. , Chih-Feng Yen M.D., Ph.D. , Hasan Alhasan M.D. , Asli Ozmen Ph.D. , Erika P. New M.D. M.P.H. , Xiaofang Guo M.D. , Anthony N. Imudia M.D. , Ozlem Guzeloglu-Kayisli Ph.D. , Charles J. Lockwood M.D. , Umit A. Kayisli Ph.D.
{"title":"Dysregulated expression of GATA2 and GATA6 transcription factors in adenomyosis: implications for impaired endometrial receptivity","authors":"Zoran Jason Pavlovic M.D. , Angel Hsin-Yu Pai M.D. , Tzu-Ti Hsiao M.S. , Chih-Feng Yen M.D., Ph.D. , Hasan Alhasan M.D. , Asli Ozmen Ph.D. , Erika P. New M.D. M.P.H. , Xiaofang Guo M.D. , Anthony N. Imudia M.D. , Ozlem Guzeloglu-Kayisli Ph.D. , Charles J. Lockwood M.D. , Umit A. Kayisli Ph.D.","doi":"10.1016/j.xfss.2023.11.003","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p><span>To study the effect of adenomyosis<span> on the localized expression of the GATA binding proteins 2 and 6 (GATA2 and GATA6) zinc-finger transcription factors that are involved in proliferation of hematopoietic and endocrine </span></span>cell lineages<span>, cell differentiation, and organogenesis, potentially leading to impaired endometrial implantation.</span></p></div><div><h3>Design</h3><p>Laboratory based experimental study.</p></div><div><h3>Setting</h3><p>Academic hospital and laboratory.</p></div><div><h3>Patients</h3><p><span>Human endometrial stromal cells (HESCs) of reproductive age patients, 18–45 years of age, with adenomyosis were compared with patients with no pathology and leiomyomatous uteri as controls (n = 4 in each group, respectively). Additionally, midsecretory phase endometrial sections were obtained from patients with adenomyosis and control patients with </span>leiomyoma (n = 8 in each group, respectively).</p></div><div><h3>Interventions</h3><p><span>GATA2<span><span> and GATA6 </span>immunohistochemistry and H-SCORE were performed on the midsecretory 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</span></span><sup>-8</sup><span> M estradiol (E2), or decidualization media (EMC) containing 10</span><sup>-8</sup> M E2, 10<sup>-7</sup><span> M medroxyprogesterone acetate, and 5 × 10</span><sup>-5</sup><span> M cAMP for 6 and 10 days. Additionally, control HESC cultures (n = 4) were transfected with scrambled small interfering RNA (siRNA) (control) or </span><em>GATA2</em>-specific siRNAs for 6 days while adenomyosis HESC cultures (n = 4) were transfected with human <em>GATA2</em><span> expression vectors to silence or induce </span><em>GATA2</em> overexpression.</p></div><div><h3>Main Outcome Measures</h3><p><span><span>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), </span>estrogen receptor 1 (ESR1), leukemia inhibitory factor (LIF), and Interleukin receptor 11 (IL11R) messenger RNA (mRNA) levels were analyzed using by qPCR with normalization to </span><em>ACTB</em><span>. 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.</span></p></div><div><h3>Results</h3><p><span><span>Immunohistochemistry revealed an overall fourfold lower GATA2 and fourfold higher GATA6 H-SCORE level in the endometrial stromal cells of patients with adenomyosis vs. controls. Decidual induction with EMC resulted in significantly lower GATA2, PGR, PRL and IGFBP1 mRNA levels in HESC cultures from patients with adenomyosis patient vs. controls. Leukemia inhibitory factor and </span>IL11R mRNA levels were also significantly dysregulated in adenomyosis HESCs compared with controls. . Silencing of </span><em>GATA2</em> expression in control HESCs induced an adenomyosis-like state with significant reductions in GATA2, increases in GATA6 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></div><div><h3>Conclusion</h3><p><span>In-vivo and in-vitro experiment results demonstrate that there is an overall inverse relationship between endometrial GATA2 and GATA6 levels in patients with adenomyosis who have diminished GATA2 levels and concurrently 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 patients with adenomyosis or </span><em>GATA2-</em>silenced control HESCs, support impaired decidualization. These effects were partially restored with <em>GATA2</em> overexpression in adenomyosis HESCs, demonstrating a potential therapeutic target.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 92-103"},"PeriodicalIF":0.0000,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"F&S science","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666335X23000691","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Objective
To study the effect of adenomyosis on the localized expression of the GATA binding proteins 2 and 6 (GATA2 and GATA6) zinc-finger transcription factors that are involved in proliferation of hematopoietic and endocrine cell lineages, cell differentiation, and organogenesis, potentially leading to impaired endometrial implantation.
Design
Laboratory based experimental study.
Setting
Academic hospital and laboratory.
Patients
Human endometrial stromal cells (HESCs) of reproductive age patients, 18–45 years of age, with adenomyosis were compared with patients with no pathology and leiomyomatous uteri as controls (n = 4 in each group, respectively). Additionally, midsecretory phase endometrial sections were obtained from patients with adenomyosis and control patients with leiomyoma (n = 8 in each group, respectively).
Interventions
GATA2 and GATA6 immunohistochemistry and H-SCORE were performed on the midsecretory 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 10-8 M E2, 10-7 M medroxyprogesterone acetate, and 5 × 10-5 M cAMP for 6 and 10 days. Additionally, control HESC cultures (n = 4) were transfected with scrambled small interfering RNA (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) messenger RNA (mRNA) levels were analyzed using 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
Immunohistochemistry revealed an overall fourfold lower GATA2 and fourfold higher GATA6 H-SCORE level in the endometrial stromal cells of patients with adenomyosis vs. controls. Decidual induction with EMC resulted in significantly lower GATA2, PGR, PRL and IGFBP1 mRNA levels in HESC cultures from patients with adenomyosis patient vs. controls. Leukemia inhibitory factor and IL11R mRNA levels were also significantly dysregulated in adenomyosis HESCs compared with controls. . Silencing of GATA2 expression in control HESCs induced an adenomyosis-like state with significant reductions in GATA2, increases in GATA6 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.
Conclusion
In-vivo and in-vitro experiment results demonstrate that there is an overall inverse relationship between endometrial GATA2 and GATA6 levels in patients with adenomyosis who have diminished GATA2 levels and concurrently 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 patients with adenomyosis or GATA2-silenced control HESCs, support impaired decidualization. These effects were partially restored with GATA2 overexpression in adenomyosis HESCs, demonstrating a potential therapeutic target.