2011-05-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/645671摘要：背景：微小RNA (microRNA)對生物體之影響是非常廣泛的，而且是較新的研究題材。我們研究團隊以過去十年之急性骨髓性白血病病人為基礎，建立了一個完整之臨床、染色體、與基因突變之資料庫。我們預期建立這群病人之微小RNA 的表達概況 (profile)，有助於了解微小RNA 在這些病人的致病角色、與各種臨床表現與基因突變之關聯性、並發展可能的治療標的。目標：我們擬在三年內完成以下目標(1) 完成約 200 位病人之微小RNA 之表達概況，以此為基礎，與這些病人之臨床、生物、與基因突變特徵相整合。(2) 找到可以影響治療反應與個人化醫療有關之微小 RNA。(3) 找尋可以影響微小 RNA 表達或功能之DNA 甲基化。(4) 找尋可以成為治療標的的微小 RNA。實驗設計與方法：目標 (1) 與 (2)：運用Applied Biosystems 即時定量聚合脢鏈鎖反應的平台，建立約二百位急性骨髓性白血病病人之微小RNA 之表達概況，並結合子計畫一之結果，希望藉由這個計畫聯結微小RNA 與急性骨髓性白血病之臨床與生物特徵。目標 (3)：我們將結合子計畫二之結果，加上生物資訊的工具，希望找到某些與微小RNA 之表達或功能有相關之DNA 甲基化。目標 (4)：我們將把可能有致病或抑病之微小RNA視為可能之治療標的，並以細胞培養與動物模型 (子計畫四) 作為驗證與進一步研討之工具。預期成果：我們可以去探討任何我們有興趣之臨床與生物特徵，並立即得到相關之微小RNAsignatures。這就好像一個寶庫，可以由我們盡情地去挖掘新的科學現象。我們將由這些微小RNA 發掘這些有趣的臨床與生物特徵之致病機轉。由於微小RNA 與DNA 甲基化有密切之互相調控，我們也將與正在進行之mRNA profile 與本整合型計劃之子計畫二之DNA 甲基化探討互相對照，預期可以發現多種新的兩者之關聯性。最後，我們預期這些新發現之微小RNA 可以是治療之新標的；有致癌性的微小RNA (oncomir)可以用locked nucleic acid (LNA)技術予以抑制；或是有抑癌性之微小RNA，可以在小鼠模型中直接以靜脈注射方式補充，而達到抑癌之效果。因此，我們將與整合型計劃之子計畫四之小鼠模型結合，驗證這些微小RNA 作為治療之可行性。<br> Abstract: Background: Since its discovery in 1993, microRNA has been found to be present from algae to humansand playing a critical role in development and tumorigenesis. By complementing with the 3’-untranslatedregion, microRNA inhibits target gene translation. Because an microRNA may have hundreds of targets,we can expect the tremendous impact of microRNA in biology. In addition, microRNA is a relatively newarea of research. We can expect fruitful and novel results by exploring this topic. In the past years, wehave established a database consisting of complete clinical, cytogenetic, and genetic information in 500acute myeloid leukemia (AML) patients. By this database, we have published dozens of papers relating toclinical and biological characterization of various genetic mutations in previous years. However, the rolesof microRNAs underlying these genetic alterations are still elusive.Specific Aims: We propose four aims in this sub-project, as the following:(1) To establish AML patients’ microRNA profiles, which will serve as a “seed” for integration with clinical,biological, and molecular data.(2) To search for microRNAs which can be helpful in risk stratification for AML and achievement ofpersonalized medicine.(3) To establish links between microRNA and epigenetic deregulations in AML.(4) To search for specific microRNA which can possibly serve as a target for treatment of AML.Design and Methods:For Aim (1) and (2) : We are now planning to measure microRNA profiles in these patients by AppliedBiosystems real-time PCR platform, followed by integration of microRNA and the previouslyestablished information and the new findings in sub-project 1.For Aim (3): We will integrate microRNA profiling with sub-project 2, methylome profiling, andcombine bioinfomatic tools to search for relationship between specific microRNA and DNAmethylation.For Aim (4): We will validate the potential therapeutic targets by in vitro cell culture system and then inanimal model in sub-project 4.Anticipated results: We expect that this combined database will facilitate us a versatile tool to sort outnovel microRNA signatures related to any phenotypes we are interested in, for example, short AMLsurvivors with NPM1 mutation but lacking FLT3-ITD mutation, a genotype thought to be a favorableprognostic factor. We believe microRNA signatures specific to these patients would be helpful to find riskmolecular factors which differentiate these unfortunate patients from the others. This database integratingclinical, biological, mRAN, methylome, and microRNA will be like a treasure, in which we can dig tosearch for many important questions. We expect to link these microRNAs with the pathogenesis pathways.Many important pathogenetic pathways have been discovered from microRNA, such as microRNA-335,which was discovered by comparison of miRNomes of breast cancer cells with and without metastasispotential, turned out to be a suppressor of metastasis, and the down-regulation of this microRNA inadvanced tumors was related to hypermethylation of the promoter. We will also compare the microRNAarray with mRNA array and methylome (sub-project 2) to establish links among these molecules, amongwhich reciprocal regulation has been frequently reported. Finally, these microRNAs may serve astherapeutic targets. By knocking down oncomir by locked nucleic acid (LNA) technology, or by augmenting tumor suppressor microRNA by direct injection into animals, tumor growth is expected to beinhibited. We will test this hypothesis in our sub-project 4, an animal model project. We believe thisproject will bring immense impact to research and treatment of AML.微小RNA急性骨髓性白血病信息RNAmicroRNAAMLmRNA