急性淋巴细胞白血病预后相关的非编码RNA调控网络和标志物研究

Adult acute lymphoblastic leukemia (ALL) is a common type of leukemia with bad and different prognoses, of which the mechanism is unclear. Based on our previous work in ALL, we plan to study the mechanism of adult ALL prognosis from the non-coding RNA (ncRNA) regulatory network, and try to find some marker molecules for prognosis. We will introduce the long non-coding RNA (lncRNA) into our previous constructed transcription factor (TF) and miRNA co-regulatory network. Thus we will construct new TF and ncRNA regulatory modules and apply them into ALL prognosis related network analysis. In this study, we will use high-throughput sequencing technology to detect the expression of coding genes and ncRNAs in multiple groups of ALL samples with different prognosis, which includes samples of untreatment, complete remission, non-remission, and recurrence. Then the specifically expressed genes and ncRNAs will be identified for the good and bad prognosis samples, respectively. And further the TF and ncRNA regulatory modules and networks for both good and bad prognosis ALL samples will be constructed. We will analyze the networks to find some key regulatory factors and modules, and discuss their roles in ALL prognosis. Finally, we will perform experiments to validate the targets in the regulatory modules and their effects in cell proliferation and migration etc. We will also detect the expression of these key genes in much more clinical samples to test their prognosis performance and discuss their potentialities as prognosis markers. This study will provide clues for the adult ALL risk stratification and improvement of the curative effect. It has potential application value and significance on disease study.

成人急性淋巴细胞白血病（ALL）总体预后不好、异质性大且机制不明。基于前期的ALL研究，本项目拟从非编码RNA（ncRNA）调控网络研究成人ALL预后的机制并筛选预后靶标。我们将在已有的miRNA和转录因子（TF）共调控网络研究方法基础上引入长非编码RNA，构建新的TF和ncRNA调控模块并应用于成人ALL预后研究中。首先通过测序鉴定多组不同预后的成人ALL样本（包括初治、缓解、未缓解和复发等）的基因表达，获得不同预后样本组特异表达的基因和ncRNA。然后分别构建不同预后样本中特异的TF和ncRNA调控模块和网络。综合分析并筛选网络中的关键调控因子和模块，探讨其在预后中的作用机制。最后验证调控因子和模块对靶基因及对细胞增殖、迁移等的影响，并在临床样本中检测其与预后的相关性及作为预后标志物的可行性。本项目为提高成人ALL预后效果奠定基础，具有重要的应用价值和学术意义。

白血病是恶性程度很高的血液肿瘤，其在儿童及青少年人中位居恶性肿瘤死亡率的首位。急性淋巴细胞白血病（Acute lymphoblastic leukemia，ALL）是白血病的常见类型，包括急性T淋巴细胞白血病和急性B淋巴细胞白血病，其预后异质性很大，其分子机制不清楚。因此，本项目旨在研究和解析不同预后效果的分子机制和发现新的预后靶标，将有利于对病人进行分层治疗，从而提高疗效、延长生存期。在本研究中，我们升级了动物转录因子数据库AnimalTFDB，整合研究了人类转录因子ChIP-Seq数据，构建了hTFtarget人类转录因子靶基因数据库，建立和升级了lncRNASNP数据库，系统分析了转录因子TF对包括lncRNA在内的靶基因的调控，以及lncRNA与miRNA的结合调控研究。针对获取的ALL病人的mRNA和miRNA表达数据和病人的预后信息。分析了ALL复发和缓解病人初治时的基因表达和miRNA表达差异，并构建了转录因子调控网络，筛选了核心调控模块和分子。在复发网络中，我们发现miR-1246/1248和转录因子WT1/SOX4/REL调控核心基因如NOTCH2的表达，另一方面，在缓解网络中，miR-429-MYCN-MFHAS1的调控模块可能起重要重要。实验验证表明，miR-1246/1248可以通过NOTCH2信号通路促进T淋细胞系的增殖。同时我们还针对K562细胞分泌的微泡可以诱导正常造型干祖细胞恶性转变为白血病细胞的机制进行了研究。通过该诱导过程多时间点样本的测序和调控网络分析，发现微泡中的转录因子和miRNA对多个癌症相关通路进行调控，同时发现miR-146b-5p调控着多个NOTCH通路的核心基因。随后我们对miR-146b-5p及其靶基因NUMB在该癌变诱导过程中的作用进行了验证，发现该miRNA可以显著促进癌症过程，NUMB可以显著延缓该过程。进一步证明miR-146b-5p可以影响基因组不稳定性和增加ROS水平。该工作证明了癌细胞的微泡可以通过携带和释放多种转录因子和miRNA等分子诱导癌症的发生。