BMP4绑定LncRNA协同组蛋白H3K4甲基化修饰调控鸡PGCs特化的机制

PGCs is the progenitor cell of all gametes, the lack of which will cause infertility in humans and animals, and it is also an important medium for the preservation of poultry resources and the production of transgenic chicken. However, since the mechanism of its development is unclear, its application has met an encountered bottleneck. PGCs is derived from ESCs, they have the same DNA genetic information but completely different biological functions, which indicated that epigenetic regulation plays an important role in PGCs development. In our previous study, we found that Lnc-PGC and H3K4me2 were involved in the differentiation of chicken ESCs into PGCs. Therefore, it was hypothesized that H3K4me2 methylation may activate or inhibit gene by regulating the transcriptional activity of Lnc-PGC，It aslo may be that Lnc-PGC acts as a scaffold for H3K4 methylase complex regulating the H3K4me2 methylation site on gene promoter to activate or inhibit gene expression. Therefore, the role of Lnc-PGC and H3K4 methylation and its relationship with target gene will be tracked by functional deletion method; Exploring the Synergistic Regulation of Lnc-PGC and H3K4me2 Methylation by Molecular Biology. The aim of this study was to reveal the mechanism of LncRNA and H3K4 modification to differentiate ESCs into PGCs, and to provide theoretical support for poultry egg production and human clinical reproductive research.

PGCs是配子的祖细胞，如有缺陷会使人和动物不育，它亦是家禽资源保存和转基因鸡制作的重要介质。但因其发育调控机制不明使其应用遇到了瓶颈。PGCs是从ESCs特化而来，两者DNA遗传信息相同但生物功能完全不同，说明表观遗传起了主要调控作用。本课题前期发现Lnc-PGC和H3K4me2参与鸡PGCs特化过程且共同靶向BMP基因和信号通路。故我们推测，可能H3K4me2甲基化修饰调控了Lnc-PGC的转录活性；也可能Lnc-PGC作为H3K4me2甲基化酶复合物的支架来调控启动子上的H3K4me2甲基化位点激活或抑制基因。故本课题拟将用功能缺失法继续追踪Lnc-PGC和H3K4me2甲基化作用及其与分化标记基因的关系；用分子生物学方法探索Lnc-PGC和H3K4me2甲基化的协同调控机制；揭示它们通过分化基因介导信号通路对ESCs分化为PGCs的双价调控作用，为家禽生产和人类生殖研究提供理论支撑

按照项目申请书及合同书的计划，本课筛选出对鸡ESCs分化为PGCs的关键LncRNA并命名为LncBMP4，进一步研究了LncBMP4和H3K4me2协同靶向BMP基因和信号通路参与鸡PGCs特化过程。该课题主要研究结果如下：（1）筛选并确定鸡原始生殖细胞形成过程中LncBMP4及其靶基因BMP4功能及其调控机制，发现LncBMP4在鸡原始生殖细胞形成过程中发挥重要作用；（2）明确原始生殖细胞形成过程中LncBMP4与BMP4的关系，确定LncBMP4通过调控BMP4来促进原始生殖细胞形成的分子机制；（3）阐明鸡原始生殖细胞形成过程中组蛋白H3K4me2甲基化与BMP4的关系，结果表明组蛋白H3K4me2甲基化与BMP4互作调控PGCs特化；（4）厘清组蛋白H3K4me2甲基化和DNA甲基化协同LncBMP4调控BMP4促进鸡原始生殖细胞特化的分子机制，阐明LncBMP4作为甲基化转移酶复合物支架参与组蛋白甲基化修饰从而调节PGCs形成的分子机制。该课题的研究按照合同书计划内容正常有序地进行，并完成全部研究计划。其研究结果解析了LnRNA和甲基化等表观遗传学因素参与鸡PGCs特化的具体分子机制，为相关家禽生产和人类生殖的相关研究提供了理论参考。