Exploiting gene expression profiling to identify novel minimal residual disease markers of neuroblastoma.
Exploiting gene expression profiling to identify novel minimal residual disease markers of neuroblastoma.
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DOI：
10.1158/1078-0432.ccr-08-0541
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发表时间：
2008-11-01
期刊：
影响因子：
--
通讯作者：
Cheung NK
中科院分区：
文献类型：
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作者：
Cheung IY;Feng Y;Gerald W;Cheung NK
微小残留病（MRD）是治疗转移性神经母细胞瘤（NB）的一个重要障碍。针对MRD的生物治疗可以改善结局。评价治疗疗效需要测量MRD，作为替代终点。由于肿瘤的异质性，没有单一的标记物可能是足够的。全基因组表达谱分析可以揭示肿瘤中与正常骨髓/血液中差异表达的潜在MRD标记物。应用AffyesterU-95基因芯片对48例4期肿瘤和9例缓解期骨髓进行基因表达谱分析。确定了34个肿瘤-骨髓表达比高于酪氨酸羟化酶的基因。对所有34个基因进行定量RT-PCR，以研究肿瘤细胞检测的动态范围以及这些基因在正常骨髓/血液样品和4期NB肿瘤中的表达。然后在2个周期的免疫治疗后，在从相同治疗方案收集的4期患者的骨髓中测试排名最高的标志物的预后意义。基于敏感性测定，鉴定了8个排名靠前的标记物：CCND 1、CRMP 1、DDC、GABRB 3、ISL 1、KIF 1A、PHOX 2B和TACC 2。它们在4期NB肿瘤（n=20）中大量表达，并且在正常骨髓/血液样品（n=20）中低至未检测到。此外，在2个治疗周期后取样的116个骨髓中CCND 1、DDC、GABRB 3、ISL 1、KIF 1A和PHOX 2B的表达是无进展生存和总生存的高度预后（p<0.001）。基于差异基因表达谱、严格的灵敏度和特异性测定以及注释良好的患者样本的标记物发现可以快速优先考虑和鉴定神经母细胞瘤的潜在MRD标记物。
Minimal residual disease (MRD) presents a significant hurdle to curing metastatic neuroblastoma (NB). Biologic therapies directed against MRD can improve outcome. Evaluating treatment efficacy requires MRD measurement, which serves as surrogate endpoint. Because of tumor heterogeneity, no single marker will likely be adequate. Genome-wide expression profiling can uncover potential MRD markers differentially expressed in tumors over normal marrow/blood. Gene expression array was carried out on 48 stage 4 tumors and 9 remission marrows using the Affymetrix U-95 gene chip. 34 genes with a tumor-to-marrow expression ratio higher than tyrosine hydroxylase were identified. Quantitative RT-PCR was performed on all 34 genes to study the dynamic range of tumor cell detection and the expression of these genes in normal marrow/blood samples and in stage 4 NB tumors. Top ranking markers were then tested for prognostic significance in the marrows of stage 4 patients collected from the same treatment protocol after 2 cycles of immunotherapy. Based on sensitivity assays, 8 top-ranking markers were identified: CCND1, CRMP1, DDC, GABRB3, ISL1, KIF1A, PHOX2B, and TACC2. They were abundantly expressed in stage 4 NB tumors (n=20) and had low to no detection in normal marrow/blood samples (n=20). Moreover, expression of CCND1, DDC, GABRB3, ISL1, KIF1A, and PHOX2B in 116 marrows sampled after 2 treatment cycles was highly prognostic of progression-free and overall survival (p<0.001). Marker discovery based on differential gene expression profiling, stringent sensitivity and specificity assays, and well-annotated patient samples can rapidly prioritize and identify potential MRD markers of neuroblastoma.