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Abstract：
BACKGROUND: Fibrogenesis within ovarian endometrioma (endometrioma), mainly induced by transforming growth factor-β (TGF-β), is characterized by myofibroblast over-activation and excessive extracellular matrix (ECM) deposition, contributing to endometrioma-associated symptoms such as infertility by impairing ovarian reserve and oocyte quality. However, the precise molecular mechanisms that underpin the endometrioma- associated fibrosis progression induced by TGF-β remain poorly understood.
METHODS: The expression level of lysine acetyltransferase 14 (KAT14) was validated in endometrium biopsies from patients with endometrioma and healthy controls, and the transcription level of KAT14 was further confirmed by analyzing a published single-cell transcriptome (scRNA-seq) dataset of endometriosis. We used overexpression, knockout, and knockdown approaches in immortalized human endometrial stromal cells (HESCs) or human primary ectopic endometrial stromal cells (EcESCs) to determine the role of KAT14 in TGF-β-induced fibrosis. Furthermore, an adeno-associated virus (AAV) carrying KAT14-shRNA was used in an endometriosis mice model to assess the role of KAT14 in vivo.
RESULTS: KAT14 was upregulated in ectopic lesions from endometrioma patients and predominantly expressed in activated fibroblasts. In vitro studies showed that KAT14 overexpression significantly promoted a TGF-β-induced profibrotic response in endometrial stromal cells, while KAT14 silencing showed adverse effects that could be rescued by KAT14 re-enhancement. In vivo, Kat14 knockdown ameliorated fibrosis in the ectopic lesions of the endometriosis mouse model. Mechanistically, we showed that KAT14 directly interacted with serum response factor (SRF) to promote the expression of α-smooth muscle actin (α-SMA) by increasing histone H4 acetylation at promoter regions; this is necessary for TGF-β-induced ECM production and myofibroblast differentiation. In addition, the knockdown or pharmacological inhibition of SRF significantly attenuated KAT14-mediating profibrotic effects under TGF-β treatment. Notably, the KAT14/SRF complex was abundant in endometrioma samples and positively correlated with α-SMA expression, further supporting the key role of KAT14/SRF complex in the progression of endometrioma-associated fibrogenesis.
CONCLUSIONS: Our results shed light on KAT14 as a key effector of TGF-β-induced ECM production and myofibroblast differentiation in EcESCs by promoting histone H4 acetylation via co-operating with SRF, representing a potential therapeutic target for endometrioma-associated fibrosis.
METHODS: The expression level of lysine acetyltransferase 14 (KAT14) was validated in endometrium biopsies from patients with endometrioma and healthy controls, and the transcription level of KAT14 was further confirmed by analyzing a published single-cell transcriptome (scRNA-seq) dataset of endometriosis. We used overexpression, knockout, and knockdown approaches in immortalized human endometrial stromal cells (HESCs) or human primary ectopic endometrial stromal cells (EcESCs) to determine the role of KAT14 in TGF-β-induced fibrosis. Furthermore, an adeno-associated virus (AAV) carrying KAT14-shRNA was used in an endometriosis mice model to assess the role of KAT14 in vivo.
RESULTS: KAT14 was upregulated in ectopic lesions from endometrioma patients and predominantly expressed in activated fibroblasts. In vitro studies showed that KAT14 overexpression significantly promoted a TGF-β-induced profibrotic response in endometrial stromal cells, while KAT14 silencing showed adverse effects that could be rescued by KAT14 re-enhancement. In vivo, Kat14 knockdown ameliorated fibrosis in the ectopic lesions of the endometriosis mouse model. Mechanistically, we showed that KAT14 directly interacted with serum response factor (SRF) to promote the expression of α-smooth muscle actin (α-SMA) by increasing histone H4 acetylation at promoter regions; this is necessary for TGF-β-induced ECM production and myofibroblast differentiation. In addition, the knockdown or pharmacological inhibition of SRF significantly attenuated KAT14-mediating profibrotic effects under TGF-β treatment. Notably, the KAT14/SRF complex was abundant in endometrioma samples and positively correlated with α-SMA expression, further supporting the key role of KAT14/SRF complex in the progression of endometrioma-associated fibrogenesis.
CONCLUSIONS: Our results shed light on KAT14 as a key effector of TGF-β-induced ECM production and myofibroblast differentiation in EcESCs by promoting histone H4 acetylation via co-operating with SRF, representing a potential therapeutic target for endometrioma-associated fibrosis.
摘要：
背景：卵巢子宫内膜瘤（子宫内膜瘤）中的纤维化，主要由转化生长因子-β（TGF-β）诱导，其特征是肌成纤维细胞过度激活和过度的细胞外基质（ECM）沉积，通过损害卵巢储备和卵母细胞质量导致子宫内膜瘤相关症状，如不孕症。然而,TGF-β诱导的子宫内膜瘤相关纤维化进展的确切分子机制尚不清楚。
方法：在子宫内膜瘤患者和健康对照的子宫内膜活检中验证赖氨酸乙酰转移酶14（KAT14）的表达水平，通过分析已发表的子宫内膜异位症的单细胞转录组（scRNA-seq）数据集，进一步证实了KAT14的转录水平。我们使用过表达，击倒,在永生化人子宫内膜基质细胞（HESCs）或人原发性异位子宫内膜基质细胞（EcESCs）中采用敲除方法，以确定KAT14在TGF-β诱导的纤维化中的作用。此外，在子宫内膜异位症小鼠模型中使用携带KAT14-shRNA的腺相关病毒(AAV)来评估KAT14在体内的作用.
结果：KAT14在子宫内膜瘤患者的异位病变中上调，主要在活化成纤维细胞中表达。体外研究表明，KAT14过表达显著促进子宫内膜基质细胞TGF-β诱导的促纤维化反应，而KAT14沉默显示了可以通过KAT14重新增强来挽救的不良反应。在体内,Kat14敲除改善子宫内膜异位症小鼠模型异位病变的纤维化。机械上,我们发现KAT14与血清反应因子（SRF）直接相互作用，通过增加启动子区域的组蛋白H4乙酰化来促进α-平滑肌肌动蛋白（α-SMA）的表达；这对于TGF-β诱导的ECM产生和肌成纤维细胞分化是必需的。此外,在TGF-β治疗下,SRF的敲减或药理学抑制显著减弱了KAT14介导的促纤维化作用。值得注意的是,KAT14/SRF复合物在子宫内膜瘤样本中含量丰富，与α-SMA表达呈正相关，进一步支持KAT14/SRF复合物在子宫内膜瘤相关纤维化进展中的关键作用。
结论：我们的结果揭示了KAT14作为TGF-β诱导的ECM产生和肌成纤维细胞分化的关键效应因子，通过与SRF协同作用促进组蛋白H4乙酰化，代表子宫内膜瘤相关纤维化的潜在治疗靶点。
方法：在子宫内膜瘤患者和健康对照的子宫内膜活检中验证赖氨酸乙酰转移酶14（KAT14）的表达水平，通过分析已发表的子宫内膜异位症的单细胞转录组（scRNA-seq）数据集，进一步证实了KAT14的转录水平。我们使用过表达，击倒,在永生化人子宫内膜基质细胞（HESCs）或人原发性异位子宫内膜基质细胞（EcESCs）中采用敲除方法，以确定KAT14在TGF-β诱导的纤维化中的作用。此外，在子宫内膜异位症小鼠模型中使用携带KAT14-shRNA的腺相关病毒(AAV)来评估KAT14在体内的作用.
结果：KAT14在子宫内膜瘤患者的异位病变中上调，主要在活化成纤维细胞中表达。体外研究表明，KAT14过表达显著促进子宫内膜基质细胞TGF-β诱导的促纤维化反应，而KAT14沉默显示了可以通过KAT14重新增强来挽救的不良反应。在体内,Kat14敲除改善子宫内膜异位症小鼠模型异位病变的纤维化。机械上,我们发现KAT14与血清反应因子（SRF）直接相互作用，通过增加启动子区域的组蛋白H4乙酰化来促进α-平滑肌肌动蛋白（α-SMA）的表达；这对于TGF-β诱导的ECM产生和肌成纤维细胞分化是必需的。此外,在TGF-β治疗下,SRF的敲减或药理学抑制显著减弱了KAT14介导的促纤维化作用。值得注意的是,KAT14/SRF复合物在子宫内膜瘤样本中含量丰富，与α-SMA表达呈正相关，进一步支持KAT14/SRF复合物在子宫内膜瘤相关纤维化进展中的关键作用。
结论：我们的结果揭示了KAT14作为TGF-β诱导的ECM产生和肌成纤维细胞分化的关键效应因子，通过与SRF协同作用促进组蛋白H4乙酰化，代表子宫内膜瘤相关纤维化的潜在治疗靶点。
