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GCgx: transcriptome-wide exploration of the response to glucocorticoids. GCgx:糖皮质激素应答的转录组范围探索。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-12-10 DOI: 10.1530/JME-21-0107
Qilin Cao, Yamil Boo Irizarry, Svetlana Yazhuk, Thai Tran, Manasi Gadkari, Luis Miguel Franco

Glucocorticoids are the cornerstone of immunosuppressive and anti-inflammatory therapy in humans, yet the mechanisms of glucocorticoid immunoregulation and toxicity remain unclear. The response to glucocorticoids is highly cell type-dependent, so translating results from different experimental systems into a better understanding of glucocorticoid effects in humans would benefit from rapid access to high-quality data on the response to glucocorticoids by different cell types. We introduce GCgx, a web application that allows investigators to quickly visualize changes in transcript abundance in response to glucocorticoids in a variety of cells and species. The tool is designed to grow by the addition of datasets based on input from the user community. GCgx is implemented in R and HTML and packaged as a Docker image. The tool and its source code are publicly available.

糖皮质激素是人类免疫抑制和抗炎治疗的基础,但糖皮质激素的免疫调节机制和毒性尚不清楚。对糖皮质激素的反应高度依赖于细胞类型,因此,将不同实验系统的结果转化为更好地理解人类对糖皮质激素的作用,将受益于快速获取不同细胞类型对糖皮质激素反应的高质量数据。我们介绍GCgx,一个网络应用程序,允许研究人员快速可视化转录物丰度的变化,以响应糖皮质激素在各种细胞和物种。该工具被设计为通过添加基于用户社区输入的数据集来增长。GCgx是用R和HTML实现的,并打包为Docker镜像。该工具及其源代码是公开的。
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引用次数: 1
Non-cell autonomous mechanisms control mitochondrial gene dysregulation in polycystic ovary syndrome. 非细胞自主机制控制多囊卵巢综合征线粒体基因失调。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-12-03 DOI: 10.1530/JME-21-0212
Alba Moreno-Asso, Ali Altıntaş, Luke C McIlvenna, Rhiannon K Patten, Javier Botella, Andrew J McAinch, Raymond J Rodgers, Romain Barrès, Nigel K Stepto

Polycystic ovary syndrome (PCOS) is a common endocrine disorder associated with insulin resistance and impaired energy metabolism in skeletal muscle, the aetiology of which is currently unclear. Here, we mapped the gene expression profile of skeletal muscle from women with PCOS and determined if cultured primary myotubes retain the gene expression signature of PCOS in vivo. Transcriptomic analysis of vastus lateralis biopsies collected from PCOS women showed lower expression of genes associated with mitochondrial function, while the expression of genes associated with the extracellular matrix was higher compared to controls. Altered skeletal muscle mRNA expression of mitochondrial-associated genes in PCOS was associated with lower protein expression of mitochondrial complex II-V, but not complex I, with no difference in mitochondrial DNA content. Transcriptomic analysis of primary myotube cultures established from biopsies did not display any differentially expressed genes between controls and PCOS. Comparison of gene expression profiles in skeletal muscle biopsies and primary myotube cultures showed lower expression of mitochondrial and energy metabolism-related genes in vitro, irrespective of the group. Together, our results show that the altered mitochondrial-associated gene expression in skeletal muscle in PCOS is not preserved in cultured myotubes, indicating that the in vivo extracellular milieu, rather than genetic or epigenetic factors, may drive this alteration. Dysregulation of mitochondrial-associated genes in skeletal muscle by extracellular factors may contribute to the impaired energy metabolism associated with PCOS.

多囊卵巢综合征(PCOS)是一种常见的内分泌疾病,与胰岛素抵抗和骨骼肌能量代谢受损有关,其病因目前尚不清楚。在这里,我们绘制了PCOS女性骨骼肌的基因表达谱,并确定培养的原代肌管在体内是否保留了PCOS的基因表达特征。从PCOS妇女收集的股外侧肌活检组织的转录组学分析显示,与线粒体功能相关的基因表达较低,而与细胞外基质相关的基因表达高于对照组。PCOS骨骼肌线粒体相关基因mRNA表达改变与线粒体复合体II-V蛋白表达降低相关,但与复合体I蛋白表达降低无关,线粒体DNA含量无差异。从活检中建立的原代肌管培养物的转录组学分析未显示对照组和多囊卵巢综合征之间有任何差异表达的基因。骨骼肌活检和原代肌管培养中基因表达谱的比较显示,体外线粒体和能量代谢相关基因的表达较低,与组无关。总之,我们的研究结果表明,PCOS骨骼肌中线粒体相关基因表达的改变在培养的肌管中没有保留,这表明体内细胞外环境,而不是遗传或表观遗传因素,可能驱动这种改变。细胞外因素对骨骼肌线粒体相关基因的失调可能导致与多囊卵巢综合征相关的能量代谢受损。
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引用次数: 4
miR-23a/b-3p promotes hepatic lipid accumulation by regulating Srebp-1c and Fas. miR-23a/b-3p通过调节Srebp-1c和Fas促进肝脏脂质积累。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-11-26 DOI: 10.1530/JME-20-0324
Linfang Li, Xiaoyi Zhang, Hangjiang Ren, Xiuqing Huang, Tao Shen, Weiqing Tang, Lin Dou, Jian Li

miR-23a-3p and miR-23b-3p are members of the miR-23~27~24-2 superfamily. The role of miR-23a/b-3p in regulating hepatic lipid accumulation is still unknown. Here, we found that increased miR-23a-3p and miR-23b-3p levels were accompanied by an increase in the protein levels of the sterol regulatory element-binding protein-1 (SREBP-1) and fatty acid synthase (FAS) in the steatotic livers of mice fed a high-fat diet and leptin receptor-deficient type 2 diabetic mice (db/db). Importantly, overexpression of miR-23a/b-3p in Hep1-6 cells elevated the intracellular triglyceride level and upregulated the expression of Srebp-1c and Fas. Taken together, these results suggested that miR-23a/b-3p enhanced mRNA stability by binding the 5'-UTR of Srebp-1c and Fas mRNA, thereby promoting triglyceride accumulation in hepatocytes.

miR-23a-3p和miR-23b-3p是miR-23~27~24-2超家族的成员。miR-23a/b-3p在调节肝脏脂质积累中的作用尚不清楚。在这里,我们发现miR-23a-3p和miR-23b-3p水平的升高伴随着高脂肪饮食小鼠和瘦素受体缺陷型2型糖尿病小鼠脂肪化肝脏中胆固醇调节元件结合蛋白-1 (SREBP-1)和脂肪酸合成酶(FAS)蛋白水平的升高(db/db)。重要的是,Hep1-6细胞中miR-23a/b-3p的过表达升高了细胞内甘油三酯水平,上调了Srebp-1c和Fas的表达。综上所述,这些结果表明miR-23a/b-3p通过结合Srebp-1c和Fas mRNA的5'-UTR增强mRNA的稳定性,从而促进甘油三酯在肝细胞中的积累。
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引用次数: 11
Downregulation of miR-216a-5p and miR-652-3p is associated with growth and invasion by targeting JAK2 and PRRX1 in GH-producing pituitary tumours. miR-216a-5p和miR-652-3p的下调通过靶向JAK2和PRRX1在gh产生的垂体肿瘤中与生长和侵袭相关。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-11-26 DOI: 10.1530/JME-21-0070
Yang Jong Lee, Chan Woo Kang, Ju Hun Oh, Jean Kim, Jong-Pil Park, Ju Hyung Moon, Eui Hyun Kim, Soohyun Lee, Se Hoon Kim, Cheol Ryong Ku, Eun Jig Lee

Expression of aberrant microRNA (miRNA) is associated with tumour formation, migration, and invasion. However, there is limited information about the epigenetics of pituitary tumorigenesis. This study investigated the role of miRNA expression during the tumorigenesis of growth hormone (GH)-secreting pituitary tumours. miRNA profiling and real-time PCR were used to analyse the mRNA expression profile in sequential pituitary tissues of a unique animal model with a GH-producing pituitary tumour. Selected miRNAs were further validated in GH-producing cell lines and human pituitary tumour samples. The expression of significantly altered miRNAs and their predicted targets, as detected by microarray, was evaluated by real-time PCR, Western blotting, and immunohistochemistry using samples from mouse models and human pituitary tumours. The effect of miRNAs on tumour proliferation and invasion was examined in GH3 cells using the MTS and Matrigel invasion assays. Among the 14 miRNAs whose expression was significantly changed, miR-216a-5p (fold change = -5.638, P -value = 0.014) and miR-652-3p (fold change = -3.482, P -value = 0.010) were constantly and significantly downregulated. Transfection with mimics of miR-216a-5p and miR-652-3p inhibited GH3 proliferation and invasion, whereas inhibitors promoted them. The direct target genes of miR-216a-5p and miR-652-3p were Jak2 and Prrx1, respectively, which were downregulated in GH3 cells transfected with mimics and in serial pituitary gland tissues, including hyperplasic tissues and tumours of acromegalic animal models and pituitary tumour tissues of acromegalic patients. Downregulated miR-216a-5p and miR-652-3p expression may contribute to tumour progression by targeting JAK2 and PRRX1 on GH-producing pituitary tumours.

异常microRNA (miRNA)的表达与肿瘤的形成、迁移和侵袭有关。然而,关于垂体肿瘤发生的表观遗传学信息有限。本研究探讨了miRNA表达在分泌生长激素(GH)的垂体肿瘤发生过程中的作用。采用miRNA谱分析和实时PCR技术分析了一种独特的垂体gh瘤动物模型序列垂体组织中mRNA的表达谱。选择的mirna在gh产生细胞系和人垂体肿瘤样本中进一步验证。通过微阵列检测到的显著改变的mirna及其预测靶标的表达,通过实时PCR、Western blotting和免疫组织化学对小鼠模型和人垂体肿瘤样本进行评估。在GH3细胞中,使用MTS和Matrigel侵袭试验检测mirna对肿瘤增殖和侵袭的影响。在14个表达发生显著变化的mirna中,miR-216a-5p (fold change = -5.638, P -value = 0.014)和miR-652-3p (fold change = -3.482, P -value = 0.010)持续且显著下调。转染miR-216a-5p和miR-652-3p模拟物可抑制GH3的增殖和侵袭,而抑制剂可促进GH3的增殖和侵袭。miR-216a-5p和miR-652-3p的直接靶基因分别为Jak2和Prrx1,在模拟物转染的GH3细胞和一系列垂体组织中下调,包括肢端肥大动物模型的增生组织和肿瘤以及肢端肥大症患者的垂体肿瘤组织。下调的miR-216a-5p和miR-652-3p表达可能通过靶向JAK2和PRRX1在gh产生的垂体肿瘤上促进肿瘤进展。
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引用次数: 5
MicroRNA-21 enhances estradiol production by inhibiting WT1 expression in granulosa cells. MicroRNA-21通过抑制颗粒细胞中WT1的表达来促进雌二醇的产生。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-11-12 DOI: 10.1530/JME-21-0162
Renée Emily Hilker, Bo Pan, Xiaoshu Zhan, Julang Li

In antral follicles, the transition of proliferative granulosa cells to estradiol-producing is critical for proper oocyte maturation. MicroRNAs are noncoding RNAs that play important roles in ovarian follicular development; however, this has yet to be fully characterized. MicroRNA-21 is significantly higher in granulosa cells isolated from large antral follicles compared to those from small antral follicles. To investigate the function of miR-21, porcine granulosa cells were transfected with miR-21 mimic or miR-21 targeted siRNA. Cells with the miR-21 mimic had higher aromatase expression and estradiol production but decreased WT1 expression. Conversely, cells with the miR-21 siRNA secreted less estradiol and had higher WT1 expression. We hypothesized that miR-21 promotes estradiol production by inhibiting WT1 protein synthesis. We found a potential miR-21 binding site in the 3'UTR of the WT1 transcript and performed a dual-luciferase reporter assay using the WT and mutated 3'UTR. Compared to the negative control, the miR-21 mimic induced a significant decrease in luciferase activity in the WT 3'UTR. This decrease was reversed when the 3'UTR was mutated, suggesting miR-21 targets this site to inhibit WT1 expression. We next transfected porcine granulosa cells with WT1 targeted siRNA and observed a significant increase in aromatase expression and estradiol secretion. We propose that miR-21 represses WT1 expression in granulosa cells to potentially promote aromatase expression and estradiol production. This study offers the first report of a microRNA regulating WT1 expression in granulosa cells and reveals the role of miR-21 in WT1's regulation of estradiol production.

在窦卵泡中,增殖颗粒细胞向产生雌二醇的转变对卵母细胞成熟至关重要。MicroRNAs是在卵巢卵泡发育中起重要作用的非编码rna;然而,这还没有得到充分的描述。MicroRNA-21在大腔滤泡分离的颗粒细胞中的表达明显高于小腔滤泡分离的颗粒细胞。为了研究miR-21的功能,我们用miR-21模拟物或miR-21靶向siRNA转染猪颗粒细胞。含有miR-21模拟物的细胞具有更高的芳香化酶表达和雌二醇产生,但降低了WT1表达。相反,含有miR-21 siRNA的细胞分泌雌二醇较少,WT1表达较高。我们假设miR-21通过抑制WT1蛋白合成来促进雌二醇的产生。我们在WT1转录本的3'UTR中发现了一个潜在的miR-21结合位点,并使用WT和突变的3'UTR进行了双荧光素酶报告基因检测。与阴性对照相比,miR-21模拟物诱导WT 3'UTR中荧光素酶活性显著降低。当3'UTR发生突变时,这种下降被逆转,这表明miR-21靶向该位点抑制WT1的表达。接下来,我们用WT1靶向siRNA转染猪颗粒细胞,观察到芳香化酶表达和雌二醇分泌显著增加。我们提出miR-21抑制颗粒细胞中WT1的表达,可能促进芳香化酶的表达和雌二醇的产生。本研究首次报道了在颗粒细胞中调控WT1表达的microRNA,揭示了miR-21在WT1调控雌二醇产生中的作用。
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引用次数: 7
GPER mediates the IL6/JAK2/STAT3 pathway involved in VEGF expression in swine ovary GCs. GPER介导参与猪卵巢GCs中VEGF表达的IL6/JAK2/STAT3通路。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-11-12 DOI: 10.1530/JME-21-0125
Longfei Xiao, Zihui Wang, Ning Lu, Yanan He, Limin Qiao, Xihui Sheng, Xiaolong Qi, Kai Xing, Yong Guo, Di Chang, Xiangguo Wang, Junjin Zhao, Xiaobin Deng, Hemin Ni, Jian Kang

Vascular endothelial growth factor (VEGF) plays a pivotal role in angiogenesis in ovaries, particularly during follicular development and ovulation. Interleukin-6 (IL-6) is one of the major pro-inflammatory factors that are involved in the angiogenesis process physiologically and pathologically. Previous studies have shown that 17β-estradiol (E2) stimulates VEGF expression by upregulating hypoxia-inducible factor 1α (HIF-1α) in many cell types, and the high level of E2 causes an inflammatory-like microenvironment before ovulation. However, whether IL-6 signaling is involved in E2-regulating VEGF expression in swine granulosa cells (GCs) is still unknown. In this study, we found the estrogen membrane receptor, G-protein-coupled estrogen receptor 1 (GPER), was expressed in swine GCs, and the expression level of GPER, HIF-1α, and VEGF increased with follicular development. In vitro study showed that E2, ICI182780, and GPER agonist (G1) promoted the expressions of HIF-1α and VEGF in swine GCs, while GPER antagonist (G15) inhibited the stimulating effect of E2 and G1. Meanwhile, G15 inhibited the stimulating effect of E2 and G1 on IL-6 mRNA expression and secretion. Furthermore, IL-6 antibody and AG490 (JAK2/STAT3 inhibitor) attenuated G1-induced HIF-1α and VEGF expression. In conclusion, this study revealed how estrogen-induced HIF-1α and VEGF expressions in swine GCs are mediated through GPER-derived IL-6 secretion leading to JAK2/STAT3 activation.

血管内皮生长因子(VEGF)在卵巢血管生成中起关键作用,特别是在卵泡发育和排卵过程中。白细胞介素-6 (IL-6)是主要的促炎因子之一,在生理和病理上参与血管生成过程。先前的研究表明,17β-雌二醇(E2)通过上调多种细胞类型中的缺氧诱导因子1α (HIF-1α)来刺激VEGF的表达,高水平的E2在排卵前引起炎症样微环境。然而,IL-6信号是否参与e2调节猪颗粒细胞(GCs)中VEGF的表达尚不清楚。本研究发现,雌激素膜受体g蛋白偶联雌激素受体1 (GPER)在猪GCs中有表达,且GPER、HIF-1α和VEGF的表达水平随卵泡发育而升高。体外研究表明,E2、ICI182780和GPER激动剂(G1)可促进猪GCs中HIF-1α和VEGF的表达,而GPER拮抗剂(G15)可抑制E2和G1的刺激作用。同时,G15抑制E2和G1对IL-6 mRNA表达和分泌的刺激作用。此外,IL-6抗体和AG490 (JAK2/STAT3抑制剂)可减弱g1诱导的HIF-1α和VEGF的表达。总之,本研究揭示了雌激素诱导的猪GCs中HIF-1α和VEGF表达是如何通过gper来源的IL-6分泌介导JAK2/STAT3激活的。
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引用次数: 5
Bone morphogenetic protein 2 controls steroid-induced osteonecrosis of the femoral head via directly inhibiting interleukin-34 expression. 骨形态发生蛋白2通过直接抑制白细胞介素-34的表达来控制类固醇诱导的股骨头骨坏死。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-10-21 DOI: 10.1530/JME-21-0163
Meng Wang, Hong Sung Min, Haojie Shan, Yiwei Lin, Wenyang Xia, Fuli Yin, Chaolai Jiang, Xiaowei Yu

Increased inflammatory response is one of the major characteristics of osteonecrosis of the femoral head (ONFH). We aimed to investigate the function of bone morphogenetic protein 2 (BMP-2)/interleukin (IL)-34 axis in the inflammatory responses of ONFH. The systemic and local expression of BMPs in ONFH patients was detected by qRT-PCR and ELISA. In vitro osteoclast differentiation and ONFH mouse models, induced by 20 mg/kg methylprednisolone through i.m. injection, were established using WT and BMP-2-/- mice to explore the regulatory role of BMP-2 in pro-inflammatory responses and bone defects of ONFH. IL-34 expression and function were examined in vitro and in vivo through qRT-PCR, tartrate-resistant acid phosphatase (TRAP) staining, and gene knockout. The systemic and local expression of BMPs was elevated in ONFH patients. BMP-2 reduced the production of pro-inflammatory cytokines and inhibited the differentiation of osteoclasts. Mechanistically, BMP-2 inhibited osteoclasts formation through suppressing IL-34 expression and then promoted bone repair and alleviated ONFH. In conclusion, our study reveals that BMP-2 inhibits inflammatory responses and osteoclast formation through downregulating IL-34.

炎症反应增加是股骨头骨坏死(ONFH)的主要特征之一。我们旨在探讨骨形态发生蛋白2 (BMP-2)/白细胞介素(IL)-34轴在ONFH炎症反应中的作用。采用qRT-PCR和ELISA检测ONFH患者bmp的全身和局部表达。采用20 mg/kg甲基强的松龙体外注射诱导破骨细胞分化和ONFH小鼠模型,以WT和BMP-2-/-小鼠为模型,探讨BMP-2在ONFH促炎反应和骨缺损中的调节作用。通过qRT-PCR、抗酒石酸酸性磷酸酶(TRAP)染色和基因敲除检测IL-34在体外和体内的表达和功能。ONFH患者的全身和局部bmp表达升高。BMP-2减少促炎细胞因子的产生,抑制破骨细胞的分化。机制上,BMP-2通过抑制IL-34表达抑制破骨细胞形成,促进骨修复,减轻ONFH。总之,我们的研究表明BMP-2通过下调IL-34抑制炎症反应和破骨细胞的形成。
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引用次数: 4
25 years of ERβ: a personal journey. 25年的ERβ:一个人的旅程。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-10-15 DOI: 10.1530/JME-21-0121
Margaret Warner, Xiaotang Fan, Anders Strom, Wanfu Wu, Jan-Åke Gustafsson

Summary: After the discovery of ERβ, a novel role for dihydrotestosterone (DHT) in estrogen signaling was revealed. Instead of just being a better androgen, DHT was found to be a precursor of the ERβ agonist 5α-androstane-3β, 17β-diol (3βAdiol), an estrogen which does not require aromatase for its synthesis. ERβ was found to oppose androgen signaling and thus is a potential target for treatment of prostate cancer. ERβ was also found to have effects that were independent of androgen signaling, particularly in the CNS. Although in rodent models of neurodegenerative diseases (Parkinson's disease, multiple sclerosis, and Alzheimer's disease), ERβ agonists are very effective in relieving symptoms and improving pathologies, this has not proven to be the case in humans. In this review we will focus on the main differences in ERβ signaling between rodents and humans and will make the point that a very important difference between the two species is in the splice variants which are expressed in humans and not rodents. The main conclusion at this point is that before we think of using ERβ agonists clinically, much more work on ERβ signaling in the human or in primates needs to be done.

摘要:ERβ被发现后,双氢睾酮(DHT)在雌激素信号传导中的新作用被揭示。DHT不仅是一种更好的雄激素,还被发现是ERβ激动剂5α-雄甾烷-3β, 17β-二醇(3β二醇)的前体,这是一种不需要芳香化酶合成的雌激素。ERβ被发现可以对抗雄激素信号传导,因此是治疗前列腺癌的潜在靶点。ERβ也被发现具有独立于雄激素信号的作用,特别是在中枢神经系统中。虽然在神经退行性疾病(帕金森病、多发性硬化症和阿尔茨海默病)的啮齿动物模型中,ERβ激动剂在缓解症状和改善病理方面非常有效,但在人类中尚未被证明是如此。在这篇综述中,我们将重点讨论啮齿动物和人类之间ERβ信号传导的主要差异,并指出两个物种之间的一个非常重要的区别是在人类而不是啮齿动物中表达的剪接变体。在这一点上的主要结论是,在我们考虑临床上使用ERβ激动剂之前,需要对人类或灵长类动物的ERβ信号传导进行更多的研究。
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引用次数: 6
Characterization of a mutated KCNJ5 gene, G387R, in unilateral primary aldosteronism. KCNJ5基因G387R在单侧原发性醛固酮增多症中的突变特征
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-09-28 DOI: 10.1530/JME-20-0282
Jeff S Chueh, Kang-Yung Peng, Vin-Cent Wu, Shuo-Meng Wang, Chieh-Kai Chan, Yung-Ming Chen, Yi-Yu Ke, Chien-Yuan Pan, Hung-Wei Liao

Somatic mutation in the KCNJ5 gene is a common driver of autonomous aldosterone overproduction in aldosterone-producing adenomas (APA). KCNJ5 mutations contribute to a loss of potassium selectivity, and an inward Na+ current could be detected in cells transfected with mutated KCNJ5. Among 223 unilateral primary aldosteronism (uPA) individuals with a KCNJ5 mutation, we identified 6 adenomas with a KCNJ5 p.Gly387Arg (G387R) mutation, previously unreported in uPA patients. The six uPA patients harboring mutant KCNJ5-G387R were older, had a longer hypertensive history, and had milder elevated preoperative plasma aldosterone levels than those APA patients with more frequently detected KCNJ5 mutations. CYP11B2 immunohistochemical staining was only positive in three adenomas, while the other three had co-existing multiple aldosterone-producing micronodules. The bioinformatics analysis predicted that function of the KCNJ5-G387R mutant channel could be pathological. However, the electrophysiological experiment demonstrated that transfected G387R mutant cells did not have an aberrantly stimulated ion current, with lower CYP11B2 synthesis and aldosterone production, when compared to that of the more frequently detected mutant KCNJ5-L168R transfected cells. In conclusion, mutant KCNJ5-G387R is not a functional KCNJ5 mutation in unilateral PA. Compared with other KCNJ5 mutations, the observed mildly elevated aldosterone expression actually hindered the clinical identification of clinical unilateral PA. The KCNJ5-G387R mutation needs to be distinguished from functional KCNJ5 mutations during genomic analysis in APA evaluation because of its functional silence.

KCNJ5基因的体细胞突变是醛固酮产生性腺瘤(APA)中自主醛固酮过量产生的常见驱动因素。KCNJ5突变导致钾选择性丧失,在转染突变KCNJ5的细胞中可以检测到向内的Na+电流。在223例KCNJ5突变的单侧原发性醛固酮增多症(uPA)患者中,我们发现了6例具有KCNJ5 p.Gly387Arg (G387R)突变的腺瘤,此前未在uPA患者中报道。6例携带突变KCNJ5- g387r的uPA患者年龄较大,高血压病史较长,术前血浆醛固酮水平升高较常见于KCNJ5突变的APA患者轻。CYP11B2免疫组化染色仅在3个腺瘤中呈阳性,而其他3个腺瘤共存多个醛固酮生成微结节。生物信息学分析预测KCNJ5-G387R突变通道的功能可能是病理性的。然而,电生理实验表明,与更频繁检测到的突变体KCNJ5-L168R转染的细胞相比,转染的G387R突变体细胞没有异常刺激的离子电流,CYP11B2合成和醛固酮产生较低。总之,突变体KCNJ5- g387r不是单侧PA的功能性KCNJ5突变。与其他KCNJ5突变相比,观察到的轻度升高的醛固酮表达实际上阻碍了临床单侧PA的临床鉴定。由于KCNJ5- g387r突变的功能沉默,在APA评估的基因组分析中需要将其与功能性KCNJ5突变区分开来。
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引用次数: 1
A20 controls expression of beta-cell regulatory genes and transcription factors. A20控制β细胞调控基因和转录因子的表达。
IF 3.5 4区 医学 Q2 ENDOCRINOLOGY & METABOLISM Pub Date : 2021-09-28 DOI: 10.1530/JME-21-0076
Wiktoria Ratajczak, Sarah D Atkinson, Catriona Kelly

TNFAIP3 encodes a zinc finger protein called A20, which has potent anti-inflammatory and anti-apoptotic properties. A20 promotes beta-cell survival and protects against islet graft rejection in experimental models. The current study sought to investigate the mechanisms underlying the protective role of A20 in the pancreatic beta-cell. Two islet cell types were used for experiments: the insulin-secreting BRIN-BD11 cell line and human islet cells. A20 was silenced using siRNA against TNFAIP3, and knockdown was confirmed by qPCR and immunostaining of cells. Cell viability, cytotoxicity and apoptosis were assessed using the ApotoxGlo assay. Glucose-stimulated insulin secretion and production of inflammatory cytokines (TNFa, IL1b and IFNg) were measured by ELISA. Expression of beta-cell regulatory genes (Abcc8, Kcnj11, Kcnq1, Gck, Scl2a2) and transcription factors (Hnf1a, Pdx1, Nkx6.1, Ngn3) was determined by qPCR. A20 deficiency increased apoptosis, impaired glucose-induced insulin secretion, and reduced expression of beta-cell regulatory genes and transcription factors. Addition of recombinant A20 normalized gene expression profiles. TNFa, IL1b and IFNg were elevated in A20 deficient cells and found to independently elicit changes in gene expression. Analysis of PCR array data suggests that A20 action in the beta cell is largely, although not exclusively, driven by the P65 subunit of NF-kB. The current report demonstrates a role for A20 in controlling beta-cell integrity and survival, which likely results from the regulation of inflammatory signalling. Of particular note is the impact that A20 deficiency has on the expression of transcription factors regulating the maturation and normal function of beta cells.

TNFAIP3编码一种叫做A20的锌指蛋白,这种蛋白具有有效的抗炎和抗凋亡特性。在实验模型中,A20促进β细胞存活并防止胰岛移植排斥反应。目前的研究旨在探讨A20在胰腺β细胞中的保护作用的机制。实验采用两种胰岛细胞:分泌胰岛素的BRIN-BD11细胞系和人胰岛细胞。A20用siRNA对TNFAIP3进行沉默,qPCR和细胞免疫染色证实A20表达下调。采用ApotoxGlo法测定细胞活力、细胞毒性和凋亡。采用ELISA法检测葡萄糖刺激胰岛素分泌及炎症因子(TNFa、IL1b、IFNg)的产生。通过qPCR检测β细胞调控基因(Abcc8、Kcnj11、Kcnq1、Gck、Scl2a2)和转录因子(Hnf1a、Pdx1、Nkx6.1、Ngn3)的表达。A20缺乏会增加细胞凋亡,损害葡萄糖诱导的胰岛素分泌,降低β细胞调节基因和转录因子的表达。添加重组A20归一化基因表达谱。在A20缺陷细胞中,发现TNFa、IL1b和IFNg升高,并独立引起基因表达的变化。PCR阵列数据分析表明,A20在β细胞中的作用主要(尽管不是完全)由NF-kB的P65亚基驱动。目前的报告表明,A20在控制β细胞完整性和存活方面发挥作用,这可能是通过调节炎症信号传导来实现的。特别值得注意的是A20缺乏对调节β细胞成熟和正常功能的转录因子表达的影响。
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引用次数: 3
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Journal of molecular endocrinology
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