Studying potential interplay between active demethylation and WT1-dependent transcriptional regulation during glial differentiation.

研究神经胶质细胞分化过程中主动去甲基化和 WT1 依赖性转录调控之间的潜在相互作用。

• 批准号: BB/N005759/1
• 负责人: Alexey Ruzov
• 金额: $ 63.98万
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FN005759%2F1
• 关键词: Studying; potential; interplay; between; active

Cellular differentiation is governed by the switching on or off of certain genes. Methylation of cytosine contributes to the regulation of gene activity. Old patterns of DNA methylation (5-methylcytosine, 5mC) are erased and new ones are established during differentiation but it is still unclear how 5mC is removed from DNA. We showed that 5mC is being specifically modified during early brain development, which leads to its subsequent removal from certain regions of DNA in neurons and glial cells, suggesting a specific mechanism for DNA demethylation. This, most likely, contributes to the reprogramming of undifferentiated cells into mature neurons and glial cells. WT1 (Wilms' tumour 1) is a protein, which, according to several reports and our preliminary data, may interact with this modified form of 5mC and may be important for brain development. The aim of this proposal is to understand the biological significance of potential interplay between WT1 and modified 5mC taking place in embryonic brain. We propose to study the mechanisms of this process using mouse embryonic stem cells (mESCs) as a model. We will differentiate mESCs into glial lineages and will identify the genes which are being modified during glial differentiation. Moreover, we will compare these genes with the genes which are regulated by WT1 during glial differentiation. Finally we will test how the depletion of WT1 and proteins involved into the 5mC modification affect glial differentiation. The proposed research program would have multiple implications for basic science, cancer research and regenerative medicine.

细胞分化是由某些基因的开启或关闭所控制的。胞嘧啶的甲基化有助于基因活性的调节。在分化过程中，旧的DNA甲基化模式（5-甲基胞嘧啶，5 mC）被清除，新的模式被建立，但仍不清楚5 mC是如何从DNA中去除的。我们发现，5 mC在早期大脑发育过程中被特异性修饰，这导致其随后从神经元和神经胶质细胞的某些DNA区域中去除，这表明DNA去甲基化的特定机制。这很可能有助于将未分化的细胞重新编程为成熟的神经元和神经胶质细胞。WT 1（Wilms'tumor 1）是一种蛋白质，根据几份报告和我们的初步数据，它可能与这种修饰形式的5 mC相互作用，可能对大脑发育很重要。该提案的目的是了解胚胎脑中WT 1和修饰的5 mC之间潜在相互作用的生物学意义。我们建议使用小鼠胚胎干细胞（mESCs）作为模型来研究这一过程的机制。我们将分化成神经胶质细胞系的胚胎干细胞，并将确定在神经胶质细胞分化过程中被修改的基因。此外，我们将这些基因与神经胶质细胞分化过程中受WT 1调控的基因进行比较。最后，我们将测试WT 1和参与5 mC修饰的蛋白质的缺失如何影响胶质细胞分化。拟议的研究计划将对基础科学、癌症研究和再生医学产生多重影响。

项目成果

Detecting and Mapping N6-Methyladenosine on RNA/DNA Hybrids.

检测和绘制 RNA/DNA 杂交体上的 N6-甲基腺苷。

• DOI: 10.1007/978-1-0716-2477-7_22
• 发表时间: 2022
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Abakir A

LINE-1 transcription in round spermatids is associated with accretion of 5-carboxylcytosine in their open reading frames.

• DOI: 10.1038/s42003-021-02217-8
• 发表时间: 2021-06-07
• 期刊: Communications biology
• 影响因子: 5.9
• 作者: Blythe MJ;Kocer A;Rubio-Roldan A;Giles T;Abakir A;Ialy-Radio C;Wheldon LM;Bereshchenko O;Bruscoli S;Kondrashov A;Drevet JR;Emes RD;Johnson AD;McCarrey JR;Gackowski D;Olinski R;Cocquet J;Garcia-Perez JL;Ruzov A
• 通讯作者: Ruzov A

Detection of Low-Abundance DNA Modifications Using Signal Amplification-Based Immunocytochemistry.

• DOI: 10.1007/978-1-0716-0876-0_14
• 发表时间: 2021
• 期刊: Methods in molecular biology
• 作者: Abdulkadir Abakir;A. Ruzov

5-formylcytosine and 5-hydroxymethyluracil as surrogate markers of TET2 and SF3B1 mutations in myelodysplastic syndrome, respectively.

5-甲酰胞嘧啶和 5-羟甲基尿嘧啶分别作为骨髓增生异常综合征 TET2 和 SF3B1 突变的替代标记。

• DOI: 10.3324/haematol.2019.224030
• 发表时间: 2020
• 期刊: Haematologica
• 影响因子: 10.1
• 作者: Gackowski,Daniel;Gawronski,Maciej;Kerr,Cassandra;Radivoyevitch,Tomas;Zarakowska,Ewelina;Starczak,Marta;Abakir,Abdulkadir;Ruzov,Alexey;Maciejewski,JaroslawP;Olinski,Ryszard
• 通讯作者: Olinski,Ryszard

Analysis of 5-Carboxylcytosine Distribution Using DNA Immunoprecipitation.

• DOI: 10.1007/978-1-0716-0876-0_24
• 发表时间: 2021
• 期刊: Methods in molecular biology
• 作者: Abdulkadir Abakir;Fahad Alenezi;A. Ruzov