Kim Patrick Y, Tan Owen, Diakiw Sonya M, Carter Daniel, Sekerye Eric O, Wasinger Valerie C, Liu Tao, Kavallaris Maria, Norris Murray D, Haber Michelle, Chesler Lou, Dolnikov Alla, Trahair Toby N, Cheung Nai-Kong, Marshall Glenn M, Cheung Belamy B
Children's Cancer Institute Australia for Medical Research, Randwick, NSW 2031, Australia.
Bioanalytical Mass Spectrometry Facility, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, NSW, Australia.
J Proteomics. 2014 Jan 16;96:1-12. doi: 10.1016/j.jprot.2013.10.032. Epub 2013 Nov 4.
The majority of patients diagnosed with neuroblastoma present with aggressive disease. Improved detection of neuroblastoma cancer cells following initial therapy may help in stratifying patient outcome and monitoring for relapse. To identify potential plasma biomarkers, we utilised a liquid chromatography-tandem mass spectrometry-based proteomics approach to detect differentially-expressed proteins in serum from TH-MYCN mice. TH-MYCN mice carry multiple copies of the human MYCN oncogene in the germline and homozygous mice for the transgene develop neuroblastoma in a manner resembling the human disease. The abundance of plasma proteins was measured over the course of disease initiation and progression. A list of 86 candidate plasma biomarkers was generated. Pathway analysis identified significant association of these proteins with genes involved in the complement system. One candidate, complement C3 protein, was significantly enriched in the plasma of TH-MYCN(+/+) mice at both 4 and 6weeks of age, and was found to be elevated in a cohort of human neuroblastoma plasma samples, compared to healthy subjects. In conclusion, we have demonstrated the suitability of the TH-MYCN(+/+) mouse model of neuroblastoma for identification of novel disease biomarkers in humans, and have identified Complement C3 as a candidate plasma biomarker for measuring disease state in neuroblastoma patients.
This study has utilised a unique murine model which develops neuroblastoma tumours that are biologically indistinguishable from human neuroblastoma. This animal model has effectively allowed the identification of plasma proteins which may serve as potential biomarkers of neuroblastoma. Furthermore, the label-free ion count quantitation technique which was used displays significant benefits as it is less labour intensive, feasible and accurate. We have been able to successfully validate this approach by confirming the differential abundance of two different plasma proteins. In addition, we have been able to confirm that the candidate biomarker Complement C3, is more abundant in the plasma of human neuroblastoma patient plasma samples when compared to healthy counterparts. Overall we have demonstrated that this approach can be potentially useful in the identification of biomarker candidates, and that further validation of the candidates may lead to the discovery of novel, clinically useful diagnostic tools in the detection of sub-clinical neuroblastoma.
大多数被诊断为神经母细胞瘤的患者表现为侵袭性疾病。在初始治疗后改善对神经母细胞瘤癌细胞的检测可能有助于对患者预后进行分层并监测复发情况。为了鉴定潜在的血浆生物标志物，我们利用基于液相色谱 - 串联质谱的蛋白质组学方法来检测TH - MYCN小鼠血清中差异表达的蛋白质。TH - MYCN小鼠在种系中携带多个拷贝的人类MYCN癌基因，并且转基因的纯合小鼠以类似于人类疾病的方式发生神经母细胞瘤。在疾病起始和进展过程中测量血浆蛋白的丰度。生成了一份包含86种候选血浆生物标志物的清单。通路分析确定这些蛋白质与补体系统相关基因存在显著关联。一种候选物，补体C3蛋白，在4周龄和6周龄的TH - MYCN（+/ +）小鼠血浆中均显著富集，并且发现在一组人类神经母细胞瘤血浆样本中与健康受试者相比有所升高。总之，我们已经证明了神经母细胞瘤的TH - MYCN（+/ +）小鼠模型适用于鉴定人类新型疾病生物标志物，并已将补体C3鉴定为用于测量神经母细胞瘤患者疾病状态的候选血浆生物标志物。
本研究利用了一种独特的小鼠模型，该模型所产生的神经母细胞瘤肿瘤在生物学上与人类神经母细胞瘤无法区分。这种动物模型有效地使得能够鉴定出可能作为神经母细胞瘤潜在生物标志物的血浆蛋白。此外，所使用的无标记离子计数定量技术显示出显著优势，因为它劳动强度较小、可行且准确。我们通过确认两种不同血浆蛋白的差异丰度成功验证了这种方法。此外，我们已经能够确认，与健康对照相比，候选生物标志物补体C3在人类神经母细胞瘤患者血浆样本的血浆中更为丰富。总体而言，我们已经证明这种方法在鉴定生物标志物候选物方面可能有用，并且对候选物的进一步验证可能会导致发现用于检测亚临床神经母细胞瘤的新型临床有用诊断工具。
