2014-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649103摘要：大約 70%到 80%的兒童 B細胞急性淋巴性白血病可以用染色體的數目和特殊的轉位來做分類，而這些分類也有臨床預後的意義。隨著基因體分析的普及，現在幾乎所有的兒童 B細胞急性淋巴性白血病都可以依據基因的變化來分類。其中有新分類的類BCR-ABL1 急性淋巴性白血病(或是稱為類費城染色體急性淋巴性白血病)，大約佔了兒童 B細胞急性淋巴性白血病的 10%左右。其中的一半有 CRLF2 這個基因的轉位並同時有 JAK1 or JAK2 基因突變。最近用全 RNA 定序或全基因定序發現非 CRLF2 轉位的類BCR-ABL1 B細胞急性淋巴性白血病有其他 kinases 或是 cytokine receptor 轉位如ABL1、ABL2、EPOR、JAK2以及 PDGFRB。知道這些是重要的，因為有越來越多的案例報告這些病人對 Tyrosine kinase inhibitors 有效。 本研究將用定量 PCR來診斷類 BCR-ABL1 兒童 B 細胞急性淋巴性白血病，目前已經有超過 20多種新的 fusion kinases。我們將設計 cDNA capture arrays來偵測這些 fusion kinases，並做藥物測試。我們並將研究為何不同的 kinases 有相同的基因表達，最後我們將探討這些不同的 partner genes 如何影響這些 kinase的功能。 本計劃的目的以及目標 (1) 診斷台灣的類 BCR-ABL1兒童 B 細胞急性淋巴性白血病並分析其預後意義。 (2) 診斷這些案例的 fusion kinases。 (3) 做這些 fusion kinases的藥物測試。 (4) 這些 fusion kinases 的致病機轉。 方法 大約有 200例兒童 B細胞急性淋巴性白血病，我們將使用 Q-RT-PCR來診斷類BCR-ABL1 B細胞急性淋巴性白血病。並使用 RT-PCR 來診斷目前已知的 fusion kinases，若為陰性，再使用 cDNA capture arrays 或是 RNA-seq來診斷。我們亦將這些novel fusion kinase clone 至MSCV，並在 Ba/F3細胞株做藥物測試。並探討這些 partner genes 如何影響這些 kinases。 預期成果 診斷台灣兒童類 BCR-ABL1 B 細胞急性淋巴性白血病，並發現新的 fusion kinase，做藥物測試，提高這類高危險群病人的存活率。 <br> Abstract: Traditionally, about 70 to 80 % of B-ALL cases were classified by chromosomal numbers and specific genetic rearrangements into prognostically relevant subgroups. With the advent of genome-wide analyses, almost all B-ALL cases can be classified genetically. A new subtype, term BCR-ABL1-like (or Philadelphia chromosome-like) ALL, accounts for nearly 10% of B-ALL cases and had a similar gene expression profiles to BCR-ABL1 ALL. Approximately half of BCR-ABL1-like ALL cases harbor CRLF2 rearrangements and concomitant JAK1/2 mutations. Recent transcriptome and whole genome sequencing (WGS) has shown that non-CRLF2-rearranged BCR-ABL1-like ALL cases harbor a diverse of genomic alterations that activate cytokine receptors and tyrosine kinases including ABL1, ABL2, EPOR, JAK2 and PDGFRB. To identify these fusions is important because there are emerging reports of responsiveness of refractory BCR-ABL1-like ALL to appropriate Tyrosine kinase inhibitor (TKI) treatment. In this project, we will make the diagnosis of BCR-ABL1-like ALL by low density array (Mullighan 2013 ASH ).The prognosis of this subtype of ALL will be analyzed. There are more than 20 novel fusion kinases detected by RNA-seq or WGS (Robert, manuscript in preparation). We will design cDNA capture arrays to target these kinase domains and sequencings to detect these fusions. In vitro drug sensitivity test will be performed for novel fusions. Finally, we will investigate why these different kinase fusions had similar gene expression profiles and how different partner genes affect these functions of these kinases. The specific Aims and Goals of this project (1) To make the diagnosis of childhood BCR-ABL1- like ALL in Taiwan and investigations of its prognostic impacts. (2) To identify the genetic alterations of these samples. (3) In vitro drug testing of kinase fusions. (4) Cellular mechanisms of these kinasese fusions. Methodology and Approaches Total around 200 patient samples will be tested for BCR-ABL1-like gene expression. There prognostic impacts will be demonstrated. For positive cases, we will use RT-PCR to detect known fusions. If all negative, capture arrays coupled with NGS or RNA-seq to find the novel kinase fusions. We will clone new fusions into MSCV and Ba/F3 cells and test in vitro drug sensitivity. Prospective results of the project We hope to identify the childhood BCR-ABL1-Like ALL in Taiwan. The clinicians might uase TKI to treat this subtype of high-risk patients and improve the outcomes.類 BCR-ABL1 兒童 B 細胞急性淋巴性白血病ABL1ABL2EPORJAK2PDGFRBfusion kinasecDNA capture arraysRNA-seqchildhood BCR-ABL1- like ALLABL1ABL2EPORJAK2PDGFRBfusion kinasecDNA capture arraysRNA-seq。