New insights into the genic and metabolic characteristics of induced pluripotent stem cells from polycystic ovary syndrome women.
New insights into the genic and metabolic characteristics of induced pluripotent stem cells from polycystic ovary syndrome women.
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对多囊卵巢综合征女性诱导多能干细胞基因和代谢特征的新见解
DOI：
10.1186/s13287-018-0950-x
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发表时间：
2018-08-09
影响因子：
7.5
通讯作者：
Yu Y
中科院分区：
文献类型：
--
作者：
Min Z;Gao Q;Zhen X;Fan Y;Tan T;Li R;Zhao Y;Yu Y
多囊卵巢综合征（PCOS）是一种常见的影响女性生育能力的内分泌和代谢紊乱。然而，由于缺乏相应的研究模型，PCOS的病理机制尚不清楚。诱导多能干细胞（iPSC）技术已被认为是产生用于疾病建模的患者特异性干细胞的手段。方法采用RNA芯片技术和实时聚合酶链反应（RT-PCR）技术分析iPSCs的mRNA丰度。利用细胞遗传学分析对iPSCs进行核型分析。采用Seahorse Bioscience XF细胞外通量分析仪测定线粒体呼吸能力和糖酵解功能。免疫荧光和RT-PCR检测ipsc相关标志物的表达。应用免疫化学方法研究了诱导多能干细胞畸胎瘤的形成。结果利用PCOS患者体细胞建立PCOS源性iPSC模型。通过RNA微阵列的综合转录谱分析，与非PCOS患者来源的iPSCs相比，PCOS患者来源的iPSCs在体外显示出代谢异常和线粒体功能障碍。其中，两个iPSC群体共有2904个基因差异表达，其中1416个基因表达上调，1488个基因表达下调（倍数变化bbb2，p< 0.01）。基因本体（Gene Ontology， GO）项富集结果显示，上调基因富集于参与三羧酸（TCA）循环、呼吸电子传递链（ETC）和糖原分解的代谢过程和线粒体活动中。另一方面，下调的基因与细胞通讯、葡萄糖运输和摄取有关。在PCOS患者来源的iPSCs和颗粒细胞中，采用RT-PCR验证了差异表达基因。PCOS患者来源的iPSCs显示线粒体呼吸能力和糖酵解功能下降（p< 0.05），但线粒体拷贝数和生物发生增加（p< 0.05）。随后，通过二甲双胍治疗多囊卵巢综合征患者来源的iPSCs，一些与葡萄糖代谢相关的基因得以恢复。同时，线粒体的ATP生成能力和PCOS患者来源的iPSCs的糖酵解能力也部分恢复到正常水平。二甲双胍对线粒体最大呼吸能力和最大糖酵解能力影响不大。结论我们测量了PCOS女性和非PCOS女性iPSCs在基因转录和线粒体呼吸功能方面的差异。PCOS患者来源的iPSCs体外代谢基因表达异常，线粒体功能障碍。本研究为研究多囊卵巢综合征的临床病因和分子机制提供了一种新的体外细胞模型。
BackgroundPolycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder that affects female fertility. However, with the lack of a corresponding research model, the pathology mechanism of PCOS is poorly understood. Induced pluripotent stem cell (iPSC) technology has been recognized as means to generate patient-specific stem cells for disease modeling.MethodsThe mRNA abundance of iPSCs was analyzed by RNA microarray and real-time polymerase chain reaction (RT-PCR). Karyotyping of iPSCs was performed with cytogenetic analysis. The mitochondrial respiration ability and glycolytic function were measured by the Seahorse Bioscience XF extracellular flux analyzer. The expression of iPSC-associated markers was identified by immunofluorescence and RT-PCR. The teratoma formation of iPSCs was studied using immunochemistry.ResultsA PCOS patient-derived iPSC model was established from somatic cells of PCOS patients. Through comprehensive transcriptional profiling analysis of the RNA microarray, PCOS patient-derived iPSCs showed metabolic abnormalities and mitochondrial dysfunction compared with non-PCOS patient-derived iPSCs in vitro. Specifically, a total of 2904 genes were differentially expressed between the two iPSC populations, of which 1416 genes were upregulated and 1488 genes were downregulated (fold change > 2,p< 0.01). Gene Ontology (GO) term enrichment results showed that upregulated genes were enriched in metabolic processes and mitochondrial activities which participated in the tricarboxylic acid (TCA) cycle, the respiratory electron transport chain (ETC), and glycogenolysis. On the other hand, the downregulated genes were related to cell communication, glucose transport, and uptake. The differentially expressed genes were verified by RT-PCR in PCOS patient-derived iPSCs and granulosa cells from PCOS patients. The PCOS patient-derived iPSCs demonstrated decreased mitochondrial respiration ability and glycolytic function (p< 0.05) but increased mitochondrial copy numbers and biogenesis (p< 0.05). Subsequently, some genes related to glucose metabolism were rescued by treating with metformin in PCOS patient-derived iPSCs. Meanwhile, the ATP production ability of mitochondria and the glycolysis ability of PCOS patient-derived iPSCs also partially returned to normal levels. However, metformin had little effect on mitochondrial maximal respiration ability and maximal glycolytic capacity.ConclusionsWe measured differences in iPSCs from women with and without PCOS in gene transcription and mitochondrial respiratory function. PCOS patient-derived iPSCs showed abnormal expression of metabolic genes and mitochondrial dysfunction in vitro.The study provides a novel cell model in vitro for studying the clinical causes and molecular mechanisms of PCOS.
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影响因子：
7.2
作者：
Wang ET;Calderon-Margalit R;Cedars MI;Daviglus ML;Merkin SS;Schreiner PJ;Sternfeld B;Wellons M;Schwartz SM;Lewis CE;Williams OD;Siscovick DS;Bibbins-Domingo K
通讯作者：
Bibbins-Domingo K
影响因子：
6.7
作者：
Santos, TA;El Shourbagy, S;St John, JC
通讯作者：
St John, JC
影响因子：
5.8
作者：
Victor, Victor M.;Rocha, Milagros;Hernandez-Mijares, Antonio
通讯作者：
Hernandez-Mijares, Antonio
DOI：
10.18632/aging.100428
发表时间：
2012-01
期刊：
Aging
影响因子：
--
作者：
Ohmine S;Squillace KA;Hartjes KA;Deeds MC;Armstrong AS;Thatava T;Sakuma T;Terzic A;Kudva Y;Ikeda Y
通讯作者：
Ikeda Y
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya