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LINE-1-mediated retrotransposition in human pluripotent stem cells: Consequences for genomic stability of hES and hiPS cells and its derivatives

人类多能干细胞中 LINE-1 介导的逆转录转座：对 hES 和 hiPS 细胞及其衍生物基因组稳定性的影响

基本信息

批准号：
198400446
负责人：
Professor Dr. Ulrich Martin
金额：
--
依托单位：
Paul-Ehrlich-Institut - Bundesinstitut für Impfstoffe und biomedizinische Arzneimittel
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2011
资助国家：
德国
起止时间：
2010-12-31 至 2016-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/198400446?language=en
关键词：
LINE mediated retrotransposition human pluripotent

项目摘要

The use of pluripotent stem cells derived from the inner cell mass of developing blastocysts (human embryonic stem cells, hESCs) or generated by reprogramming of fibroblasts (human induced pluripo-tent stem cells, hiPSCs) holds great therapeutic promise for regenerative medicine. However, a vital unanswered question is whether these cells or their derivatives are truly safe for administration. Spe-cifically, it is unclear whether the integrity of the genome of pluripotent stem cells is maintained during their generation, expansion and differentiation. The appearance of genetic mutations during their ex-pansion or differentiation could undermine stem cell therapies. Such mutations could be induced by human non-LTR retrotransposons (LINE1 or L1, Alu, SVA) which represent the currently mobilized group of human endogenous retroelements. About 35% of the human genome are the consequence of the mobilization of non-LTR retrotransposons which is executed by the L1-encoded protein machin-ery. We will investigate whether hESCs and hiPSCs differ in their support for L1 retrotransposition and analyze the extent of genomic destabilization of these cells by L1- retrotransposition. We want to determine both the role of L1 activity in the formation of karyotypic abnormalities that are frequently observed in pluripotent stem cells in tissue culture, and the effect of differentiation on L1 mobilization rates. We will evaluate if there are any L1 integration preferences for specific genomic regions in hESC and hiPSC lines that could potentially affect neighbouring gene expression. The proposed ap-proaches will help to elucidate whether L1-mediated retrotransposition can contribute to genome fluid-ity and variability of hESCs and hiPSCs, and will be useful to assess the activity of endogenous mobile elements in pluripotent stem cells in order to evaluate their safety for therapy.

使用源自发育囊胚内细胞团（人胚胎干细胞，hESC）或通过成纤维细胞重编程产生的多能干细胞（人诱导多能干细胞，hiPSC）为再生医学带来了巨大的治疗前景。然而，一个尚未解答的重要问题是这些细胞或其衍生物是否真的可以安全使用。具体来说，尚不清楚多能干细胞在其生成、扩增和分化过程中是否保持基因组的完整性。在其扩增或分化过程中出现的基因突变可能会破坏干细胞疗法。这种突变可以由人类非 LTR 逆转录转座子（LINE1 或 L1、Alu、SVA）诱导，这些逆转录转座子代表了当前动员的人类内源性逆转录因子组。大约 35% 的人类基因组是由 L1 编码的蛋白质机器执行的非 LTR 逆转录转座子动员的结果。我们将研究 hESC 和 hiPSC 在支持 L1 逆转录转座方面是否存在差异，并分析 L1 逆转录转座对这些细胞的基因组不稳定的程度。我们想要确定 L1 活性在组织培养的多能干细胞中经常观察到的核型异常形成中的作用，以及分化对 L1 动员率的影响。我们将评估 hESC 和 hiPSC 系中特定基因组区域是否存在任何可能影响邻近基因表达的 L1 整合偏好。所提出的方法将有助于阐明L1介导的逆转录转座是否有助于hESC和hiPSC的基因组流动性和变异性，并将有助于评估多能干细胞中内源性移动元件的活性，以评估其治疗的安全性。