2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/648896摘要：用單核苷酸多態性（SNP）陣列來做全基因的掃瞄，已成功地確定了兒童急性淋巴細胞白血病（ALL）的血癌細胞存在DNA拷貝數異常（copy number abnormalities, CNAs）和雜合性缺失（loss of heterozygosity, LOH）的異常。1幾個確定的基因異常，包括在IKZF1基因缺失 (deletion)，JAK基因突變，以及IL15多態性，已成為新的預後指標。2-4我們的初步努力用multiplex PCR配合毛細管電泳系統來偵測IKZF1基因缺失，在兒童B細胞急性淋巴細胞白血病患者確認IKZF1基因缺失是一個預後不良因子。與此同時，我們也用單核苷酸多態性陣列檢驗一部份病人的檢體，並確定了在許多染色體區域有基因的缺損或是放大。另一方面，我們和校園合作的基因體核心實驗室 ， 初步成果顯示臺灣的兒童白血病具有特定的微核醣核酸(microRNA)變化影響到病人的預後（請參閱初步研究）。有了這些基礎，我們提出本研究計劃，研究以下事項：1. 台灣兒童急性淋巴細胞白血病的基因變異 。2. 希望由單核苷酸多態性陣列發現新的預後指標，宿主因素，來確定高危險群病人3. 研究微核醣核酸表達與淋巴球發育的重要基因，如PAX5，IKZF1，EBF1，和Notch1之間的關係。為了實現第一個目標：了解台灣兒童急性淋巴細胞白血病基因變異，我們會在第一年用單核苷酸多態性陣列完成共100-120白血病檢體。每個病人將包括白血病的DNA和個別病人的germline DNA，並根據其臨床結果建立預後指標。第二個目標是第二個財政年度，我們將繼續執行毛細管電泳系統檢測IKZF1缺失和用高分辨率熔解（HRM）來偵測JAK2突變熱點（R683G）。將來應用這些技術，做為單核苷酸多態性陣列驗證。我們將用另外4個醫療中心200個病人檢體，以驗證我們所找到新的預後因子。在最後一年的研究，我們將結合微核醣核酸陣列和單核苷酸多態性陣列去探討微核醣核酸表達量的改變在這幾個PAX5，IKZF1，EBF1和Notch1重要基因有變異的時候。我們將利用染色質免疫沉澱（ChIP）來驗證結果。最終我們的研究目標是增進台灣兒童急性淋巴細胞白血病風險導向治療以及發現台灣病童獨有的宿主因素。<br> Abstract: Genome-wide study by single-nucleotide polymorphism (SNP) arrays has successfully identified the presence of DNA copy number abnormalities (CNAs) and loss of heterozygosity (LOH) in childhood acute lymphoblastic leukemia (ALL).1 Several identified gene abnormalities, including deletions in the IKZF1 gene, the JAK mutations, and variants of IL15, have become new prognostic markers in association with treatment response.2-4 Our preliminary efforts by multiplex PCRs coupled with Capillary Electrophoresis System has identified IKZF1 gene deletion in our childhood B-cell ALL patients and we have also validated IKZF1 deletion to be a marker for poor prognosis. Meanwhile, we have also profiled a small set of our patients by more advanced SNP arrays and identified amplifications or deletions in many chromosomal regions. On the other hand, we have collaborated with the Microarray Core on campus and identified the microRNA expression signatures for childhood ALL by microRNA arrays (Please refer to Preliminary studies). With these, we propose this project to study the followings:(1) Genetic alterations underlying the childhood ALL in Taiwan(2) New prognostic markers, host factors, or a gene signature based on SNP arrays to identify high-risk patients(3) The up- and down-regulated microRNAs in association with the expression of developmentally important genes i.e., PAX5, IKZF1, EBF1, and Notch1.To achieve the first aim: genetic alterations underlying childhood ALL in Taiwan, we will profile a total of 100-120 leukemic samples by SNP arrays in the first fiscal year. The sample will include leukemic DNA and remission germline DNA of individual patients and the prognostic signature will be constructed according to their clinical outcome. To the second aim for the second fiscal year, we will continue to perform multiplex PCRs by Capillary Electrophoresis System to detect IKZF1 deletion and JAK2 mutation hot spot (R683G) by High Resolution Melting (HRM) and apply these techniques to the following gene signature validation. With the initial success in the construction of prognostic signature, in the second year of this proposal, we will use another 200 childhood ALL samples from another 4 medical centers to validate the prognostic signature. In the last fiscal year, we will combine microRNA arrays and SNP arrays to study the different expressions of miRNAs with aberrations of PAX5, IKZF1, EBF1 and Notch1. We will also use Chromatin immunoprecipitation (CHiP) to validate the results. The ultimate goals of our study are to apply risk-directed therapy and detect host factors unique to childhood ALL patients in Taiwan.兒童急性淋巴性白血病單核?酸多態性陣列CREBBP deletionsABDchildhood acute lymphoblastic leukemiasingle-nucleotide polymorphism arrayCREBBP deletionsABD