cAMP/PKA通路介导的恶性胶质瘤细胞诱导分化的miRNA组学研究

Differentiation therapy of malignant glioma is a novel strategy for glioma therapy which has scientific significance and promising prospects.Our previous study demonstrate that cholera toxin induces malignant glioma cell differentiation via cAMP/PKA pathway.Furthermore, we find that miR-335 is closely associated with cholera toxin-induced glioma cell differentiation, indicating the pivotal role of miRNAs in differentiation therapy. These basic researches were published in PNAS and in Molecular Pharmacology as Accelerated Communication. Nevertheless,the mechanism how miRNAs mediate glioma cell differentation induced by cAMP/PKA agonist has yet not been systematacially elucidated. In this programme, we plan to acquire the full scale of miRomes in cAMP/PKA agonist-induced glioma cell differentiation by in-depth analysis of miRomes. For aberrantly expressed miRNAs, gain and loss of function of miRNAs experiments are designed to determine if they mediate the glioma cell differentiation. And then, to disclose action modes of those key miRNAs,we will focus on their target genes and signal pathways.Most importantly, miRNA mimics or inhibitors will be applied in animal model of malignant glioma to assess the therapeutic potential of miRNA. We believe that this programme will help us to reveal molecular mechanisms of differeniation-inducing therapy for malignant glioma from Epigenetics perspective, and may provide new targets for glioma differentiation therapy.

恶性胶质瘤的诱导分化疗法是一个具有科学意义和应用前景的治疗新策略。申请者前期工作发现，霍乱毒素通过激活cAMP/PKA通路诱导恶性胶质瘤细胞的分化； 进一步研究证实，miRNA-335的上调与之密切相关，提示miRNA在恶性胶质瘤诱导分化中的重要调控作用。这些基础研究已发表于PNAS和Mol Pharm。然而，恶性胶质瘤诱导分化疗法的miRNAs控制机理尚未得到系统阐述。因此，本项目拟用miRNA深度测序对cAMP/PKA激动剂诱导的恶性胶质瘤细胞分化模型进行miRNA组学分析，以获得全面的miRNA转录组信息；再采用miRNA功能获得与缺失实验双向证明表达变化最明显的miRNAs是否介导诱导分化，并进一步探究其介导分化的靶基因和信号通路；最后在胶质瘤动物模型上验证miRNA能否作为诱导分化治疗靶点。本项目的完成有望从表观遗传学角度解读恶性胶质瘤诱导分化疗法的分子机制，并为临床提供新靶点。

恶性胶质瘤的诱导分化疗法是一个具有科学意义和应用前景的治疗新策略。申请者前期工作发现，霍乱毒素通过激活cAMP/PKA 通路诱导恶性胶质瘤细胞的分化； 进一步研究证实，miRNA-335 的上调与之密切相关，提示miRNA 在恶性胶质瘤诱导分化中的重要调控作用。这些基础研究已发表于PNAS 和Mol Pharm。然而，恶性胶质瘤诱导分化疗法的miRNAs控制机理尚未得到系统阐述。因此，本项目用miRNA 深度测序对cAMP/PKA 激动剂诱导的恶性胶质瘤细胞分化模型进行miRNA 组学分析，获得了全面的miRNA转录组信息；采用miRNA 功能获得与缺失实验,证明表达变化超过3倍的miRNAs 中miR-1275可以逆转cAMP/PKA 激动剂引起的增殖抑制和诱导分化；研究进一步发现，胶质瘤的诱导分化是由线粒体生成引起的代谢模式转变（从有氧酵解转为有氧氧化）引起，miR-1275参与线粒体生成；最后在胶质瘤动物模型上验证cAMP/PKA 激动剂可成功诱导分化并抑制胶质瘤生长。本项目的完成从表观遗传学角度解读恶性胶质瘤诱导分化疗法的分子机制，并为临床提供新靶点。