MicroRNA对胚胎干细胞的多能性网络调控

胚胎干细胞(Embryonic stem cells, ESCs)是一类能够无限增殖和诱导分化为多种类型细胞的干细胞。MicroRNA(miRNA)是一类内源性具有调控基因表达功能的非编码RNA, 在ESCs增殖和分化过程中起重要作用。MiRNA可以通过对ESCs多能性网络中的转录因子、细胞周期、表观遗传学、信号转导等方面调控, 促使ESCs维持多能性状态。文章重点综述了miRNA的生成过程、调控ESCs多能性的主要miRNA家族以及miRNA对ESCs多能性网络调控作用等内容。     

p300通过homeobox结构域增强Nanog的转录激活活性

胚胎干细胞(ES细胞)能够不断地进行自我更新来维持其多能性,很多转录因子共同调控着ES细胞的自我更新和多能性,Nanog就是其中之一,然而Nanog维持ES细胞多能性的机制并不清楚。p300是真核生物中普遍存在的转录辅助因子,与许多转录因子共同作用调控下游基因的表达。为探索p300是不是能够影响Nanog的转录活性,我们在细胞中共转Nanog(或突变体)及报告基因和p300,结果表明p300能够通过homeobox结构域增强Nanog的转录激活活性。     

维持胚胎干细胞生物学特性的分子调控机制

胚胎干(embryonic stem,ES)细胞来源于植入前的胚胎.在体外培养条件下,可保持分化成机体各种类型细胞的能力,即全能性.因此,胚胎干细胞被广泛用作研究胚胎发生、发育的细胞模型,也是目前开展细胞移植性治疗研究的重要来源.揭示维持ES细胞自我更新和全能性的分子调控机制,是ES细胞基础研究和临床治疗基础研究的重要领域.目前研究发现,不同的信号通路、转录因子及其通过激活ES细胞特异性表达谱对维持ES细胞多能性、自我更新发挥关键作用。     

调控胚胎干细胞自我更新的关键转录因子研究进展

胚胎干细胞（embryonic stem cells,ES细胞）具有自我更新和发育多能性的特点,在再生医学研究中有着广泛的应用前景。ES细胞多能性和自我更新的维持受到复杂的调控,涉及到转录调控、信号转导以及表观遗传调控等多个方面。转录因子Oct4、Sox2、Nanog在其中扮演着非常重要的角色,对干细胞特性的维持必不可少。本文着重讨论了这些关键转录因子的研究进展。这些研究促进了对ES细胞自我更新机制的深入理解,并为进一步的临床研究提供了理论基础。     

维持胚胎干细胞多能性和自我更新的转录因子Oct-4/ Nanog以及相关的调控网络

Oct-4和Nanog是两种维持干细胞多能性和自我更新的转录因子, 它们通过结合靶基因调控区, 选择性地抑制分化基因表达或促进多能性基因表达。它们通常只在多能干细胞中表达, 在分化细胞中不表达。在不同的发育阶段, 它们的表达量受到特异调控, 并且分别与Sox-2、FoxD3等其他转录因子以及LIF、BMP等胞外信号通路互相作用, 形成一个复杂的转录调节crosstalk网络, 在特异时空激活或抑制靶基因的转录; 通过互相制约最终决定干细胞是保持多能性还是分化, 以及向哪个方向分化。此外, Oct-4和Nanog对体细胞重编程为多能细胞也有重要作用。     

Rex-1对Oct4转录活性的调控

Rex-1和Oct4是在多能性细胞中特异表达的转录因子,而Rex-1的生物学功能及其调控胚胎干细胞(ES细胞)多能性和分化能力的机制尚不清楚。实验探讨了Rex-1和Oct4的相互关系,利用免疫荧光实验和免疫共沉淀实验证明了Rex-1和Oct4两种蛋白共同定位于细胞核中,证明二者之间有直接的相互作用。 进一步的活性分析表明Rex-1能够抑制Oct4的转录激活活性。这些数据提供了一种新的调控Oct4活性的机制。     

转录因子Oct-4调控下游靶基因及其与胚胎发育全能/多能性的关系

作为一个和胚胎发育全能／多能性相关的转录因子,Oct-4通过多种多样的调控机制激活或抑制不同靶基因的转录,从而在细胞的全能／多能性及未分化状态的调控维持中发挥重要的作用。已知受Oct-4调控的靶基因中,不仅有一些重要的转录因子如Rex-I,而且有一些参与重要细胞活动的基因如Fgf-4,因此对Oct-4调控下游靶基因的研究将有助于对其在分化发育中所起作用的进一步了解,同时对全能／多能性这一发育学基本问题及其调控网络有一个新的认识。     

胚胎干细胞自我复制及亚全能分化特性维持的分子机制

微环境中的各种细胞因子与细胞内的多种转录因子通过一定的信号系统相互作用构成网络, 共同调控胚胎干细胞的自我复制. 本文对维持胚胎干细胞(embryonic stem cells, ESCs)亚全能分化特性的相关分子、信号通路以及它们的作用方式进行了综述, 探讨了胚胎干细胞自我复制及亚全能分化特性维持的分子机制.     

MicroRNAs与胚胎干细胞研究

MicroRNAs(miRNAs)是一种大小约20～25个碱基的非编码小分子RNA,一般通过特异性抑制靶蛋白翻译或降解靶基因mRNA发挥负调控基因表达的作用.胚胎干细胞(embryonic stem cells,ES细胞)是从植入前早期胚胎内细胞团或原始生殖细胞中分离得到并能在体外长期培养的高度未分化的多能细胞系,在揭示胚胎早期发育机理、药物筛选、临床再生医学等领域具有广泛的应用前景.最近研究发现miRNAs在ES细胞自我更新和分化过程中均发挥着重要的调控作用,但具体调控机制尚未完全阐明.进一步深入研究miRNAs在ES细胞中的作用,全面了解ES细胞自我更新和定向分化的机制是实现ES细胞广阔临床应用前景的基础.     

MicroRNA调控动物脂肪细胞分化研究进展

脂肪组织不仅在维持机体能量代谢和稳态上发挥重要作用,同时也是重要的内分泌器官。脂肪细胞分化是由间充质干细胞(Mesenchymal stem cells, MSC)向成熟脂肪细胞分化的复杂生理过程,该过程由大量转录因子、激素、信号通路分子协同调控。miRNA作为内源性非编码RNA,主要通过抑制转录后翻译等机制来调控基因表达。近年来越来越多的证据表明miRNA通过调控脂肪细胞分化相关的转录因子和重要信号分子进而影响动物脂肪细胞的分化和脂肪形成。本文对miRNA影响动物白色、棕色和米色脂肪细胞分化的作用机制及其相关调控通路和关键因子进行了归纳总结,以期为肥胖等代谢性疾病的治疗提供一定的理论指导和新的治疗思路。     

Nanog基因的功能与调控研究进展

胚胎干细胞的无限增殖能力和亚全能性决定了它在再生医学、新药开发及发育生物学基础研究中具有巨大的应用前景。探索维持胚胎干细胞亚全能性的因子及其网络的调控功能成为胚胎干细胞生物学研究的热点。已研究发现多个与维持胚胎干细胞亚全能性相关的基因如Oct4, Nanog, Sox2等,其中Nanog是2003年5月末发现的一个基因,它对维持胚胎干细胞亚全能性起关键性作用,能够独立于L1F/Stat3维持ICM和胚胎干细胞的亚全能性。几年来,Nanog的生物学功能及其与 Oct4, Sox2等亚全能性维持基因之间的相互作用关系已有较为深入的研究,并发现多个调控Nanog表达的转录因子,从而进一步明晰Nanog与已知调控胚胎发育的信号通路之间的关系。本文在综述Nanog基因的表达特征和功能的基础上、重点探讨Nanog基因表达调控以及Oct4, Sox2等亚全能性维持基因之间的相互作用关系,并对未来的研究趋势予以展望。     

家畜胚胎干细胞多能性候选信号通路及分子标志

家畜胚胎干细胞具有重要的生物学意义和广阔的应用前景。以下对比了小鼠、人胚胎干细胞多能性调控信号通路的异同,阐述了小鼠、人胚胎干细胞与家畜胚胎干细胞在多能性分子标志上的差异,并结合本实验室开展绵羊胚胎干细胞研究的实际经验,对目前家畜胚胎干细胞建系中可能存在的多能性候选信号通路及分子标志进行了探讨。     

Nanog overcomes reprogramming barriers and induces pluripotency in minimal conditions

Induced pluripotency requires the expression of defined factors and culture conditions that support the self-renewal of embryonic stem (ES) cells. Small molecule inhibition of MAP kinase (MEK) and glycogen synthase kinase 3 (GSK3) with LIF (2i/LIF) provides an optimal culture environment for mouse ES cells and promotes transition to naive pluripotency in partially reprogrammed (pre-iPS) cells. Here we show that 2i/LIF treatment in clonal lines of pre-iPS cells results in the activation of endogenous Nanog and rapid downregulation of retroviral Oct4 expression. Nanog enables somatic cell reprogramming in serum-free medium supplemented with LIF, a culture condition which does not support induced pluripotency or the self-renewal of ES cells, and is sufficient to reprogram epiblast-derived stem cells to naive pluripotency in serum-free medium alone. Nanog also enhances reprogramming in cooperation with kinase inhibition or 5-aza-cytidine, a small molecule inhibitor of DNA methylation. These results highlight the capacity of Nanog to overcome multiple barriers to reprogramming and reveal a synergy between Nanog and chemical inhibitors that promote reprogramming. We conclude that Nanog induces pluripotency in minimal conditions. This provides a strategy for imposing naive pluripotency in mammalian cells independently of species-specific culture requirements.     

NPR-A regulates self-renewal and pluripotency of embryonic stem cells

Self-renewal and pluripotency of embryonic stem (ES) cells are maintained by several signaling cascades and by expression of intrinsic factors, such as Oct4, Nanog and Sox2. The mechanism regulating these signaling cascades in ES cells is of great interest. Recently, we have demonstrated that natriuretic peptide receptor A (NPR-A), a specific receptor for atrial and brain natriuretic peptides (ANP and BNP, respectively), is expressed in pre-implantation embryos and in ES cells. Here, we examined whether NPR-A is involved in the maintenance of ES cell pluripotency. RNA interference-mediated knockdown of NPR-A resulted in phenotypic changes, indicative of differentiation, downregulation of pluripotency factors (such as Oct4, Nanog and Sox2) and upregulation of differentiation genes. NPR-A knockdown also resulted in a marked downregulation of phosphorylated Akt. Furthermore, NPR-A knockdown induced accumulation of ES cells in the G1 phase of the cell cycle. Interestingly, we found that ANP was expressed in self-renewing ES cells, whereas its level was reduced after ES cell differentiation. Treatment of ES cells with ANP upregulated the expression of Oct4, Nanog and phosphorylated Akt, and this upregulation depended on NPR-A signaling, because it was completely reversed by pretreatment with either an NPR-A antagonist or a cGMP-dependent protein kinase inhibitor. These findings provide a novel role for NPR-A in the maintenance of self-renewal and pluripotency of ES cells.     

Identification of potential pluripotency determinants for human embryonic stem cells following proteomic analysis of human and mouse fibroblast conditioned media

The unique pluripotential characteristic of human embryonic stem cells heralds their use in fields such as medicine, biotechnology, biopharmaceuticals, and developmental biology. However, the current availability of sufficient quantities of embryonic stem cells for such applications is limited, and generating sufficient numbers for downstream therapeutic applications is a key concern. In the absence of feeder layers or their conditioned media, human embryonic stem cells readily differentiate to form embryoid bodies, indicating that trophic factors secreted by the feeder layers are required for long-term proliferation and maintenance of pluripotency. Adding further complexity to the elucidation of the factors required for the maintenance of pluripotency is the variability of different fibroblast feeder layers (of mouse or human origin) to effectively support human embryonic stem cells. Currently, the deficiency of knowledge concerning the exact identity of factors within the pathways for self-renewal illustrates that a number of factors may be required to support pluripotent, undifferentiated growth of human embryonic stem cells. This study utilized a proteomic analysis (multidimensional chromatography coupled to tandem mass spectrometry) to isolate and identify proteins in the conditioned media of three mitotically inactivated fibroblast lines (human fetal, human neonatal, and mouse embryonic fibroblasts) used to support the undifferentiated growth of human embryonic stem cells. One-hundred seventy-five unique proteins were identified between the three cell lines using a 相似文献