Non-coding RNA In Stem Cell Biology

项目摘要 多能干细胞在早期胚胎发育过程中是必不可少的，并具有巨大的潜力， 治疗应用。在这两种情况下，正确调节这些细胞的分裂和分化是至关重要的 为了一个有利的结果。目前，我们对干细胞如何维持其功能的理解存在重大差距。 多能状态，或者向分化过渡。几种类型的非编码RNA富集在 干细胞，并已提出在这些调节过程中发挥作用，特别是通过影响 细胞的染色质状态。我们将利用大量的成人多能干细胞，即新生细胞， 的Planarian Schmidtea meditriquea，以解决两种类型的非编码RNA的作用和机制， 在体内干细胞生物学中。该项目的一个分支涉及长链非编码RNA（lncRNA）在 多能性和分化。然而，许多体外筛选表明lncRNA在细胞凋亡中的作用。 尽管存在围绕多能性的调节，但已经证明了很少的体内作用的情况。我们将使用in 体内筛选真涡虫干细胞功能，首先鉴定功能上重要的lncRNA，然后将其 进一步研究机械。这将使我们能够集中精力研究lncRNA最相关的例子- 介导的干细胞调节。该项目的另一个分支重点关注PIWI-RNA相互作用的作用 在干细胞生物学中，PIWI蛋白质是PIRNA（piRNA）及其结合伴侣PIWI蛋白质的一种。这些小RNA主要是 已知它们在动物生殖系中的转座子沉默中的作用，但也是干细胞功能所必需的， 特别是在再生动物中。我们的目标是确定该要求的基础是什么，通过确定 干细胞生物学的哪些阶段受到影响以及皮尔纳途径失败的后果是什么。 此外，皮尔纳功能的几个方面在生殖细胞中不容易解决，我们将使用 Planarian干细胞，以促进我们对皮尔纳生物学的这些机制方面的理解。这个项目 将为非编码RNA的功能和机制提供新的见解，并将对我们的研究产生广泛的影响。 了解多能干细胞和一般基因组稳定性的调节。这些见解 对于多能细胞在治疗应用中的安全使用以及对于靶向治疗 与内源性干细胞功能障碍相关的病症，包括不育症、出生缺陷和衰老。 此外，该项目还涉及与细胞身份丧失有关的疾病，如癌症。

PROJECT SUMMARY Pluripotent stem cells are essential during early embryonic development, and hold great potential for therapeutic applications. In both cases, correct regulation of division and differentiation of these cells is crucial for a favorable outcome. Currently, there are major gaps in our understanding of how stem cells maintain their pluripotent state, or make the transition to differentiation. Several types of non-coding RNA are enriched in stem cells, and have been proposed to function in these regulatory processes, in particular by affecting the chromatin state of the cells. We will leverage a prolific population of adult pluripotent stem cells, the neoblasts of the planarian Schmidtea mediterranea, to address the role and mechanism of two types of non-coding RNA in in vivo stem cell biology. One branch of this project concerns the role of long non-coding RNAs (lncRNAs) in pluripotency and differentiation. Whereas many in vitro screens have suggested a role for lncRNAs in the regulation surrounding pluripotency, few cases of in vivo effects have been demonstrated. We will use an in vivo screen of planarian stem cell function to first identify functionally important lncRNAs, which will then be studied further mechanistically. This will allow us to focus our efforts on the most relevant examples of lncRNA- mediated stem cell regulation. The other branch of this project focuses on the role of PIWI-interacting RNAs (piRNAs), and their binding partners the PIWI proteins, in stem cell biology. These small RNAs are mostly known for their role in transposon silencing in the animal germline, but are also essential for stem cell function, especially in regenerating animals. We aim to determine what the basis for that requirement is, by determining which stages of stem cell biology are affected and what the consequences of failure of the piRNA pathway are. Furthermore several aspects of piRNA function are not easily addressed in germ cells, and we will use the planarian stem cells to further our understanding of those mechanistic aspects of piRNA biology. This project will provide new insights in non-coding RNA function and mechanism, and will have broad impact on our understanding of pluripotent stem cells and regulation of genomic stability in general. These insights are essential for the safe use of pluripotent cells in therapeutic applications, as well as for targeted treatment of conditions related dysfunction of endogenous stem cells including forms of infertility, birth defects, and aging. Furthermore, this project has relevance to conditions related to loss of cell identity, such as cancer.