Analyzing DOT1L-dependent hippocampus development using single-cell high throughput sequencing

使用单细胞高通量测序分析 DOT1L 依赖性海马发育

• 批准号: 449569068
• 负责人: Professorin Dr. Tanja Vogel
• 金额: --
• 依托单位: Anatomie II: Abteilung für Molekulare Embryologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/449569068?language=en
• 关键词: Analyzing; DOT1L; dependent; hippocampus; development

Epigenetic processes are implicated in central nervous system development and function. However, there is still limited insight into the role of individual chromatin modifiers and their targets in physiological and patho-physiological conditions in the central nervous system. It is largely unknown how chromatin modifications impact on hippocampus development. The histone methyltransferase Disruptor of Telomeric Silencing 1-Like (DOT1L) confers histone H3 lysine 79 (H3K79) methylations. Our preliminary data indicate that studying the function of DOT1L will give unique opportunity to narrow the gap of knowledge of how this epigenetic modifier affects hippocampal development and function. Importantly, our data indicate that DOT1L deficiency interferes with the correct exertion of fundamental processes during hippocampus development. We observe altered development of the dentate gyrus (DG) as well as the cornu ammonis (CA) fields. Our current insights suggest that the derailed developmental program is reflected by disturbed neuronal differentiation that occurs prematurely and confers incorrect cell identities.We here propose to extend a preliminary single-cell RNA data set from embryonic day (E) 16.5 control and DOT1L-deficient mouse hippocampus to further developmental time points, spanning from E12.5 until birth. Characterising the emerging data set covering various stages of development shall allow gaining comprehensive knowledge about differentiation trajectories from diverse, but partly unknown stem cell populations to the neurons of the hippocampus. Further insights into molecular alterations underlying derailed transcriptional programs in DOT1L deficient hippocampus shall come from studying H3K79 methylation and accessibility of regulatory regions, especially enhancers. We propose to use a diverse repertoire of bioinformatics analyses to explore the emerging comprehensive data set to 1) decipher the impact of H3K79 methylation in control of transcriptional programs necessary to specify the heterogeneous neuronal subpopulation, 2) infer transcriptional programs that contribute to hippocampal fissure and DG formation, 3) resolve different subsets of neural stem cells contributing to differentiation of DG and CA fields, and 4) determine gene activities needed and fine-tuned at the boundary of the subiculum and the CA neuroepithelium. We aim to decipher master regulators that determine differentiation of the diverse neurons in the hippocampus and these candidate genes shall be functionally validated in vivo through gain- or loss-of-function experiments.

表观遗传过程与中枢神经系统的发育和功能有关。然而，仍然有有限的洞察个别染色质修饰剂和它们的目标在中枢神经系统的生理和病理生理条件下的作用。染色质修饰如何影响海马发育在很大程度上是未知的。端粒沉默1样组蛋白甲基转移酶破坏因子（DOT1L）赋予组蛋白H3赖氨酸79（H3K79）甲基化。我们的初步数据表明，研究DOT1L的功能将提供独特的机会，缩小这种表观遗传修饰剂如何影响海马发育和功能的知识差距差距。重要的是，我们的数据表明，DOT1L缺乏干扰海马发育过程中基本过程的正确发挥。我们观察到改变发展的齿状回（DG）以及角ammonis（CA）领域。我们目前的见解表明，脱轨的发展计划是反映了干扰神经元分化，过早发生，并赋予不正确的细胞idents.We在这里建议延长一个初步的单细胞RNA数据集从胚胎天（E）16.5控制和DOT1L缺陷小鼠海马进一步发展的时间点，从E12.5直到出生。表征涵盖不同发育阶段的新兴数据集应允许获得关于从不同但部分未知的干细胞群体到海马神经元的分化轨迹的全面知识。DOT1L缺陷海马中出轨转录程序的分子改变的进一步见解将来自研究H3K79甲基化和调节区域，特别是增强子的可及性。我们建议使用生物信息学分析的多样性来探索新兴的综合数据集，以1）破译H3K79甲基化在控制指定异质神经元亚群所必需的转录程序中的影响，2）推断有助于海马裂和DG形成的转录程序，3）解析有助于DG和CA区分化的神经干细胞的不同子集，以及4）确定在下托和CA神经上皮的边界处所需和微调的基因活性。我们的目标是破译主调节器，确定不同的神经元在海马和这些候选基因的分化应在体内通过获得或丧失功能的实验进行功能验证。