CN111965289A - 尿液中髓母细胞瘤的代谢标志物及其用途 - Google Patents
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Abstract
本发明涉及尿液中髓母细胞瘤的代谢标志物及其用途;具体地,本发明涉及检测受试者尿液中代谢物水平的试剂在制备用于诊断和/或预后髓母细胞瘤的试剂盒和/或芯片中的用途,其中所述的代谢物选自以下中的一种或多种:尿硫蝶呤、20‑氧‑白三烯E4、皮酮四醇、四氢可的松。
Description
技术领域
本发明涉及生物技术领域,更具体地涉及尿液中髓母细胞瘤的代谢标志物及其在早期诊断和术后病情监测中的用途。
背景技术
髓母细胞瘤(Medulloblastoma,MB)是儿童中最常见的恶性中枢神经系统肿瘤(参见Brandes,A.A.and M.K.Paris,Review of the prognostic factors inmedulloblastoma of children and adults.Crit Rev Oncol Hematol,2004.50(2):p.121-8;Saran,F.,Recent advances in paediatric neuro-oncology.Curr OpinNeurol,2002.15(6):p.671-7),占儿童脑肿瘤的8%~10%(参见Archer,T.C.,E.L.Mahoney,and S.L.Pomeroy,Medulloblastoma:Molecular Classification-BasedPersonal Therapeutics.Neurotherapeutics,2017.14(2):p.265-273)。早发现,早治疗可有效提高病患的五年生存率(参见Ramaswamy,V.等人,Recurrence patterns acrossmedulloblastoma subgroups:an integrated clinical and molecularanalysis.Lancet Oncol,2013.14(12):p.1200-7;Packer,R.J.等人,Phase III study ofcraniospinal radiation therapy followed by adjuvant chemotherapy for newlydiagnosed average-risk medulloblastoma.J Clin Oncol,2006.24(25):p.4202-8;Packer,R.J.,Risk-adapted craniospinal radiotherapy followed by high-dosechemotherapy and stem-cell rescue in children with newly diagnosedmedulloblastoma.Curr Neurol Neurosci Rep,2007.7(2):p.130,132;和Tarbell,N.J.等人,High-risk medulloblastoma:a pediatric oncology group randomized trial ofchemotherapy before or after radiation therapy(POG9031).J Clin Oncol,2013.31(23):p.2936-41)。临床常采用的MB诊断方法主要用动态对比增强计算机断层扫描(CT)和磁共振成像(MRI)(Cassia,G.S.E.等人,Childhood Medulloblastoma Revisited.TopMagn Reson Imaging,2018.27(6):p.479-502)。目前的方法对一些小的髓母细胞瘤很难确认。此外,即使由经验丰富的放射科医师反复检查图像,对于MB和脑部良性疾病、其他脑部恶性肿瘤的区分也具有一定的难度(参见Bennett,C.D.等人,Tissue metaboliteprofiles for the characterisation of paediatric cerebellar tumours.Sci Rep,2018.8(1):p.11992)。因此,发展新的、特异性的、非侵入性的诊断方法对MB的早期临床干预及治疗具有重要的意义,同时也可以减少对其他脑部疾病的不必要的治疗。
代谢组学是发现疾病潜在生物标志物的有效策略(参见Srivastava,A.andD.J.Creek,Discovery and Validation of Clinical Biomarkers of Cancer:A ReviewCombining Metabolomics and Proteomics.Proteomics,2019.19(10):p.e1700448)。前期大量的代谢组研究对脑肿瘤的代谢组进行了很好的表征(参见Pandey,R.等人,Metabolomic signature of brain cancer.Mol Carcinog,2017.56(11):p.2355-2371),对MB的研究也越来越多。2010年,Cuellar-Baena,S等人应用核磁共振技术呈现了脑肿瘤组织样品的代谢谱,包括室管膜瘤、MB等。他们的研究结果揭示了MB组的代谢特征,包括膜周转增加、神经元活力降低和糖酵解改变(参见Cuellar-Baena,S.等人,Comparativemetabolic profiling of paediatric ependymoma,medulloblastoma and pilocyticastrocytoma.Int J Mol Med,2010.26(6):p.941-8)。2018年,Christopher D.Bennett等人使用NMR技术定量分析了小脑室管膜瘤、MB和其他脑肿瘤组织样本的代谢组,研究结果显示MB组中的磷酸胆碱和牛磺酸水平显著升高(参见Bennett,C.D.等人,Tissue metaboliteprofiles for the characterisation of paediatric cerebellar tumours.Sci Rep,2018.8(1):p.11992)。
前期研究表明肿瘤组织中的代谢物可以用于MB的诊断。到目前为止,尚缺乏对MB体液标志物的研究。研究发现体液代谢物可用区其他脑部肿瘤的早期诊断,提示对于MB的诊断和监测具有较大的潜力。2011年,Moroz,J等人利用小鼠模型分析了胶母细胞瘤(GBM)鼠和对照组小鼠尿液的代谢物含量。结果显示尿液中显著变化的代谢物数量在携带肿瘤的群体中比在对照动物中更多,表明代谢组学可以用作小鼠中生长的GBM细胞的监测工具(参见Moroz,J.等人,Tumour xenograft detection through quantitative analysis ofthe metabolic profile of urine in mice.Phys Med Biol,2011.56(3):p.535-56)。2016年,Zhao,H.,等应用87名胶质瘤患者进行了血浆代谢组分析。结果发现了29种差异代谢产物,其中6种代谢物可有效区分IDH1突变阳性和阴性胶质瘤患者,准确率为94.4%,以上结果表明代谢物可有效区分肿瘤表型,可能对胶质瘤的分子分型具有重要的应用价值(参见Zhao,H.等人,Metabolomics profiling in plasma samples from gliomapatients correlates with tumor phenotypes.Oncotarget,2016.7(15):p.20486-95)。
根据以往研究,应用体液标志物预警和检测MB具有潜在的临床价值。因此,本研究的目的是发现MB早期诊断及术后病情监测的尿液代谢标志物。具体地,本发明人基于MB和健康对照、脑部良性肿瘤和脑部其他恶性肿瘤的尿液代谢组学的比较,发现MB特异性诊断标志物,并在外部样本中进行验证。使用接受者操作特征(ROC)分析评估生物标记物的诊断潜力。此外,对以上MB特异性标志物进行术后含量趋势变化分析,筛选可用于MB术后病情监测的潜在标志物。
发明内容
鉴于本领域的上述需求,根据本公开的一些实施方案,提供了检测尿液中代谢物水平的试剂在制备用于诊断和/或监测髓母细胞瘤预后的试剂中的用途,其中所述的代谢物选自以下一种或多种:
尿硫蝶呤(Urothion);
20-氧-白三烯E4(20-Oxo-leukotriene E4);
皮酮四醇(Cortolone);和
四氢可的松(Tetrahydrocortisone)。
在具体的实施方案中,所述诊断是指髓母细胞瘤的早期诊断;所述监测髓母细胞瘤预后是指监测髓母细胞瘤的术后病情恢复。
在具体的实施方案中,分别相较于健康对照、脑部良性肿瘤和脑部其他恶性肿瘤的患者,髓母细胞瘤患者的尿液中尿硫蝶呤、20-氧-白三烯E4、皮酮四醇和四氢可的松的含量升高;其中健康对照是指未患有髓母细胞瘤及其他疾病的健康个体,脑部其他恶性肿瘤是指非髓母细胞瘤的其他恶性肿瘤。
在具体的实施方案中,通过质谱鉴定检测尿液中代谢物的水平,其中所述质谱鉴定是在质谱全扫描模式联合靶向分析下使用的。质谱全扫描模式是同时采集质量范围100m/z到1000m/z内的所有小分子一级信息,通过多元统计分析筛选差异代谢物,进一步对差异代谢物进行靶向二级碎裂,结合数据库二级谱图,最终确定差异代谢物。
在具体的实施方案中,检测尿液中代谢物相对于对照样本的相对水平,其中所述对照样本源自健康对照、脑部良性肿瘤和脑部髓母细胞瘤以外的脑部其他恶性肿瘤的患者。
根据本发明的具体实施方案,其中脑部良性疾病选自挛型脑性瘫痪、脑电图检查、面神经、不明原因抽搐、癫痫、肢体疼痛、共济失调反射保留型、头晕、感染、蛛网膜囊肿、癫痫复杂部分性发作继发全面、椎管内占位性病变、头痛、肌肉萎、癫痫全面性发作强直阵挛性发作、脱髓鞘脑病、癫痫单纯部分性发作、部分性癫痫、膜髓鞘病、运动障碍、脑瘫、难治性癫痫和脑性瘫痪(包括但不限于脑瘫,脑梗死,透明隔囊肿,癫痫复杂部分性发作);
另一方面,根据本发明的具体实施方案,其中脑部髓母细胞瘤以外的脑部其他恶性肿瘤选自室管膜瘤、间变性室管膜瘤、皮样囊肿、脉络丛乳头状瘤、脉络丛癌、星形细胞瘤、卵黄囊瘤、神经节细胞胶质瘤、间变型室管膜瘤、毛细胞型星形细胞瘤、混合性神经元胶质肿瘤、胶质母细胞瘤、未分化的室管膜瘤、混合性胶质神经元肿瘤、室管膜下巨细胞星形细胞瘤和菊形团形成的胶质神经元肿瘤。
在具体的实施方案中,所述受试者是人。
本发明的另一方面提供了一种用于早期诊断髓母细胞瘤和/或监测髓母细胞瘤预后的试剂盒或芯片,其包含检测以下代谢物水平的试剂:尿硫蝶呤、20-氧-白三烯E4、皮酮四醇和四氢可的松。
在具体的实施方案中,根据本发明的试剂盒或芯片用于质谱鉴定的方法中。
在具体的实施方案中,所述诊断选自:髓母细胞瘤的早期诊断;所述病情监测选自髓母细胞瘤的术后病情监测。在具体的实施方案中,所述质谱鉴定是在全扫描模式联合靶向鉴定模式下使用的。
本发明的另一方面提供了一种用于在受试者中诊断髓母细胞瘤和/或监测髓母细胞瘤预后的方法,包括步骤:
1)获得受试者的尿液样本,
2)任选地,从尿液样本中提取代谢物,
3)确定受试者尿液样本中一种或多种选自以下的代谢物的水平:尿硫蝶呤、20-氧-白三烯E4、皮酮四醇、四氢可的松及其组合。
在具体的实施方案中,使用质谱方法确定尿液样本中代谢物的水平。
当采用质谱方法确定尿液中代谢物的水平时,在获得尿液样本的步骤之后,还可以包括代谢物提取,蛋白去除。在具体的实施方案中,用2倍体积乙腈提取尿液样本中的代谢物,同时去除蛋白。
在具体的实施方案中,所述质谱方法是一级全扫描模式联合靶向二级分析。具体地,首先通过一级全扫描检测尿液代谢组,多元统计分析筛选出潜在标志物,对潜在标志物进行靶向二级碎裂,结合数据库二级谱图确定潜在标志物。采用标志物一级谱峰面积进行定量。
具体地,检测尿硫蝶呤、20-氧-白三烯E4、皮酮四醇和四氢可的松中的一种或多种。
术语
本文中的术语“尿硫蝶呤”是一种正常的钼辅助因子代谢物。它是一种从人类尿液中分离出的含硫的黄色蝶啶衍生物。研究显示,钼辅脢缺乏症患者尿液中尿硫蝶呤含量缺乏(Johnson,J.L.and Rajagopalan,K.V.(1982)Structural and metabolicrelationship between the molybdenum cofactor and urothione.Proc Natl Acad SciU S A 79(22),6856-60)。
本文中使用的术语“20-氧-白三烯E4”是白三烯E4(LTE4)通过脂质氧化得到的代谢物。白三烯E4(LTE4)是一种半胱氨酰白三烯。半胱氨酰白三烯(CysLTs)是一个强大的炎症介质家族。LTE4是由肝脏中的谷氨酰转肽酶和颗粒二肽酶催化白三烯C4得到的代谢物。(Mizutani,N.(2003)[Studies on the experimental allergic rhinitis induced byJapanese cedar pollen--role of cysteinyl leukotrienes in nasal allergicsymptoms].Yakugaku Zasshi 123(1),1-8)。
本文中的术语“皮酮四醇”是一种甾体代谢物。在孕妇(36-40周妊娠)和未怀孕妇女的尿液中以及接近足月妊娠妇女的羊水中都可以发现(Kraan,G.P.et al.(1980)Quantification of polar glucocorticosteroids in the urine of pregnant andnonpregnant women:a comparison with 6alpha-hydroxylated metabolites ofcortisol in neonatal urine and amniotic fluid.J Clin Endocrinol Metab 51(4),754-8)。
本文中的术语“四氢可的松”是一种皮质类固醇激素,参与压力应激;他能增加血压和血糖水平,抑制免疫(Stiefel,P.et al.(2002)Role of ketoconazole treatment inurinary-free cortisol-to-cortisone and tetrahydrocortisol-to-tetrahydrocortisone ratios in nonectopic Cushing's syndrome.Endocrine 18(3),279-84)。
附图说明
图1A和图1B分别显示髓母细胞瘤与健康对照组PCA得分图、和OPLS-DA得分图;
图2A和图2B分别显示髓母细胞瘤与脑部良性疾病PCA分类图、及OPLS-DA得分图;
图3A和图3B分别表示髓母细胞瘤与其他脑部恶性肿瘤PCA分类图、及OPLS-DA得分图;
图4表示皮酮四醇和四氢可的松两种代谢物组合在预测实验组样本的ROC曲线;
图5A至图5D分别表示4种代谢产物:尿硫蝶呤、20-氧-白三烯E4、12-氧-2,3-dinor-10,15-植物二烯酸、皮酮四醇、四氢可的松在健康组(对照)、髓母细胞瘤术前组(MB)和术后一月组(PO)的含量变化。
具体实施方式
实施例1:髓母细胞瘤早期特异性诊断标志物研究
以下结合附图,通过实施例进一步说明本发明,但不作为对本发明的限制。以下提供了本发明实施方案中所使用的具体材料及其来源。然而,应当理解的是,这些仅仅是示例性的,并不意图限制本发明,与如下试剂和仪器的类型、型号、品质、性质或功能相同或相似的材料均可以用于实施本发明。下述实施例中所使用的实验方法如无特殊说明,均为常规方法。下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
我们用液相色谱-高分辨质谱(LC-MS)通过全扫描模式检测尿液中代谢物,通过多元统计分析筛选两组间差异代谢物认为与髓母细胞瘤相关。标志物的鉴定是采用二级靶向分析方法,通过对二级碎片的匹配或者解析进行。
材料与试剂
1)仪器:Waters H-class液相色谱仪(waters公司)LTQ-Orbitrap velos pro质谱仪(Thermofisher Scientific公司)。
2)主要试剂:乙腈(Thermofisher Scientific公司);C18反相色谱柱(3.0mm×100mm,C18,1.7μm,Waters公司)。
3)样本:实验组:74例(男44例,女30例)髓母细胞瘤患者的术前尿,76例(男43例,女33例)年龄、性别匹配的正常对照组的尿液,34例(男24例,女10例)脑部良性疾病(包括挛型脑性瘫痪、脑电图检查、面神经、不明原因抽搐、癫痫、肢体疼痛、共济失调反射保留型、头晕、感染、蛛网膜囊肿、癫痫复杂部分性发作继发全面、椎管内占位性病变、头痛、肌肉萎、癫痫全面性发作强直阵挛性发作、脱髓鞘脑病、癫痫单纯部分性发作、部分性癫痫、膜髓鞘病、运动障碍、脑瘫、难治性癫痫、脑性瘫痪(包括脑瘫、脑梗死、透明隔囊肿、癫痫复杂部分性发作))组的尿液和20例(男10例,女10例)其他脑部恶性肿瘤(包括室管膜瘤、间变性室管膜瘤、皮样囊肿、脉络丛乳头状瘤、脉络丛癌、星形细胞瘤、卵黄囊瘤、神经节细胞胶质瘤、间变性室管膜瘤、间变型室管膜瘤、毛细胞型星形细胞瘤、混合性神经元胶质肿瘤、胶质母细胞瘤、未分化的室管膜瘤、混合性胶质神经元肿瘤、室管膜下巨细胞星形细胞瘤、菊形团形成的胶质神经元肿瘤)组的尿液;验证组:38例(男23例,女15例)髓母细胞瘤患者的术前尿液,42例(男24例,女18例)年龄、性别匹配的正常对照组的尿液,17例(男13例,女4例)脑部良性疾病组的尿液和11例(男6例,女5例)其他脑部恶性肿瘤组的尿液,来自北京天坛医院。各组平均年龄不超过2岁。
1.1人尿液样品的收集
收集空腹晨尿,5000g的转速离心30min,去除沉淀。
1.2代谢物提取
取200ul尿液上清,加200ul乙腈,涡旋,4度静置30min,14000g离心10min,取上清,离心浓缩,用200ul 2%乙腈水复溶,14000g离心10min,过10kD滤膜后取10ul进样。
1.3液相分析
Waters H-class液相色谱仪,色谱柱:waters HSS T3 C18(3.0X100mm,1.7um),柱温50℃;流动相A为0.1%甲酸水,流动相为乙腈;分析梯度为:0-1min,2%B;1-8min,2%B-98%B;8-8.1min,98%B-100%B;8.1-12min,100%B;12-12.1min,100%-2%B;12.1-17min,2%B;流速为0.5ml/min;进样体积为10ul。
1.4质谱分析
UPLC质谱串联LTQ-Orbitrap velos(Thermo Fisher Scientific,SanJose,CA,USA)质谱,采用电喷雾离子源正离子模式;鞘气为氮气和辅助气,流速分别为45arbitraryunits and 10arbitrary units;质谱扫描范围为100–1000m/z;spray voltages设为4.2KV;离子传输管温度350℃。数据采用高分辨傅里叶转换模式(FT)获取,一级分辨率为60000;二级分辨率为15000。
1.5质谱数据分析
由UPLC-LTQ orbitrap获得的原始数据,采用Waters公司的商业组学分析软件progenesis QI(Version 2.0,Nonlinear Dynamics,UK)进行处理。该软件可自动完成峰对齐,峰识别和峰校正等前处理程序,最终输出三维矩阵,即由保留时间和精确质荷比组成的谱峰索引变量、样本名称和峰强度/面积组成。获得的数据矩阵导入多变量统计软件SIMCA-P software 14.0(Umetrics AB,Umea,Sweden)进行PCA分析,可视化组间变化趋势。组间差异变量通过OPLS-DA模型获取的VIP值进行筛选,VIP值大于1,非参检验p值小于0.05的变量认为是组间显著性差异变量,筛选为髓母细胞瘤潜在标志物。对筛选的差异变量进行二级碎裂,采用HCD(High collision dissociation)碎裂方式,根据具体代谢物选择20,40,60eV能量。将二级碎片采用progenesis QI软件进行解卷积,搜索HMDB(HUMAN METABOLOMEDATABASE)数据库,确定差异代谢物结构。
结果
1.尿液代谢组区分髓母细胞瘤和健康对照组
非监督PCA得分图显示(图1A)实验组髓母细胞瘤组和健康对照组呈现出一定的区分度。进一步采用监督OPLS-DA构建模型,两组区分度更加明显(图1B)。最终筛选出53个代谢物在实验组和验证组均有显著差异。进一步应用ROC曲线评估差异代谢物对髓母细胞瘤和健康对照组的区分效果。结果显示有46个代谢物的曲线下面积(AUC)均大于0.7,提示对髓母细胞瘤均有较好的预测价值。
2.尿液代谢组区分髓母细胞瘤和脑部良性疾病组
非监督PCA得分图显示(图2A)实验组髓母细胞瘤组和脑部良性疾病组呈现出一定的区分度。进一步采用监督OPLS-DA构建模型,两组区分度更加明显(图2B)。最终筛选出40个代谢物在实验组和验证组均有差异。进一步应用ROC曲线评估差异代谢物对髓母细胞瘤和脑部良性疾病组的区分效果。结果显示有23个代谢物的曲线下面积(AUC)均大于0.7,提示对髓母细胞瘤均有较好的预测价值。
3.尿液代谢组区分髓母细胞瘤和其他脑部恶性肿瘤组
非监督PCA得分图显示(图3A)实验组髓母细胞瘤组和其他脑部恶性肿瘤组呈现出一定的区分度。进一步采用监督OPLS-DA构建模型,两组区分度更加明显(图3B)。最终筛选出26个代谢物在实验组和验证组均有差异。进一步应用ROC曲线评估差异代谢物对髓母细胞瘤和其他脑部恶性肿瘤组的区分效果。结果显示有12个代谢物的曲线下面积(AUC)均大于0.7,提示对髓母细胞瘤均有较好的预测价值。
综合分析以上1、2和3中的分析结果,我们发现有14个代谢物在以上3个实施例中对髓母细胞瘤均具有较好的预测效果(ROC>0.7),但是,通过进一步的文献调研,我们删除了尚未在人源样品中有明确报道的代谢物10种,这些代谢物可能是由于饮食等原因导致出现在尿液样本中,最后仅保留了已经明确报道为人尿液代谢物的4种,他们的检测水平恒定在一个范围内。我们所选的4种在健康人群中是比较稳定的,检测水平恒定在一定的范围内,具有作为标志物的潜力。
见表1,2、3。我们认为这4个尿液代谢物可作为髓母细胞瘤早期诊断的特异性标志物。
鉴于多个代谢物联合预测可取得更好的效果,结合生物学功能,我们发现2个代谢物,皮酮四醇和四氢可的松联合可有效区分髓母细胞瘤和非髓母细胞瘤组(包括健康对照组、脑部良性疾病组和其他脑部恶性肿瘤组)。采用随机森林算法构建ROC曲线,其曲线下面积可达0.858(图4)。
表1、4个潜在标志物区分髓母细胞瘤和健康对照
表2、4个潜在标志物区分髓母细胞瘤和良性脑肿瘤
表3、4个潜在标志物区分髓母细胞瘤和其他脑部恶性肿瘤组
实施例2:髓母细胞瘤术后病情监测标志物研究
材料与试剂
1)仪器:Waters H-class液相色谱仪(waters公司)LTQ-Orbitrap velos pro质谱仪(Thermofisher Scientific公司)。
2)主要试剂:乙腈(Thermofisher Scientific公司);C18反相色谱柱(3.0mm×100mm,C18,1.7μm,Waters公司)。
3)样本:112例(男67例,女45例)髓母细胞瘤患者的术前尿液样本,80例(男51例,女29例)髓母细胞瘤患者的术后一月的尿液样本;118例(男67例,女51例)年龄、性别匹配的正常对照组的尿液;来自北京天坛医院。各组平均年龄不超过2岁。
2.1人尿液样品的收集
收集空腹晨尿,5000g的转速离心30min,去除沉淀。
2.2代谢物提取
取200ul尿液上清,加200ul乙腈,涡旋,4度静置30min,14000g离心10min,取上清,离心浓缩,用200ul 2%乙腈水复溶,14000g离心10min,过10kD滤膜后取10ul进样。
2.3液相分析
Waters H-class
色谱柱:waters HSS T3 C18(3.0X100mm,1.7um),柱温50℃;流动相A为0.1%甲酸水,流动相为乙腈;分析梯度为:0-1min,2%B;1-8min,2%B-98%B;8-8.1min,98%B-100%B;8.1-12min,100%B;12-12.1min,100%-2%B;12.1-17min,2%B;流速为0.5ml/min;进样体积为10ul。
2.4质谱分析
UPLC质谱串联LTQ-Orbitrap velos(Thermo Fisher Scientific,SanJose,CA,USA)质谱,采用电喷雾离子源正离子模式;鞘气为氮气和辅助气,流速分别为45arbitraryunits and 10arbitrary units;质谱扫描范围为100–1000m/z;spray voltages设为4.2KV;离子传输管温度350℃。数据采用高分辨傅里叶转换模式(FT)获取,一级分辨率为60000;二级分辨率为15000。
2.5数据分析
由UPLC-LTQ orbitrap获得的原始数据,采用Waters公司的商业组学分析软件progenesis QI(Version 2.0,Nonlinear Dynamics,UK)进行处理。该软件可自动完成峰对齐,峰识别和峰校正等前处理程序,最终输出三维矩阵,即由保留时间和精确质荷比组成的谱峰索引变量、样本名称和峰强度/面积组成。从获得的数据中提取实施例1得出的4个髓母细胞瘤相关的代谢物的含量,比较健康组、术前组和术后一月组这4个特异性标志物的含量变化。筛选术后含量显著趋近于正常组的代谢物(术后vs.术前:p值小于0.05),认为是髓母细胞瘤术后病情监测的潜在标志物。
结果
髓母细胞瘤术后病情监测标志物的筛选
4个髓母细胞瘤早期诊断标志物在健康组、髓母细胞瘤术前组和术后一月组的含量变化如图5A至图5D所示,结果显示,4个代谢物的含量在术后一月较术前发生显著变化,并且术后含量明显趋近于正常对照组,说明这些代谢物均可做为髓母细胞瘤术后病情检测的潜在标志物。
Claims (10)
1.检测受试者尿液中代谢物水平的试剂在制备用于诊断髓母细胞瘤的试剂盒和/或芯片中的用途,其中所述的代谢物选自以下中的一种或多种:尿硫蝶呤、20-氧-白三烯E4、皮酮四醇和四氢可的松。
2.根据权利要求1所述的用途,其中所述诊断是指髓母细胞瘤的早期诊断。
3.根据权利要求1所述的用途,其中所述诊断可用于区分髓母细胞瘤患者与健康受试者、脑部良性肿瘤和脑部髓母细胞瘤以外的其他恶性肿瘤的患者。
4.根据权利要求1-3中任一项所述的用途,其中相较于健康对照、脑部良性肿瘤和脑部其他恶性肿瘤的患者,如果受试者尿液中的代谢物水平具有以下一种或多种变化,则诊断所述受试者患有髓母细胞瘤:尿硫蝶呤、20-氧-白三烯E4、皮酮四醇和四氢可的松的含量升高;其中健康对照是指未患有髓母细胞瘤及其他疾病的健康个体。
5.根据权利要求1-3中任一项所述的用途,其中通过质谱鉴定检测受试者尿液中的代谢物水平。
6.根据权利要求1-3中任一项所述的用途,其中,所述受试者是人。
7.检测受试者尿液中代谢物水平的试剂在制备用于监测髓母细胞瘤患者预后的试剂盒和/或芯片中的用途,其中所述的代谢物选自以下中的一种或多种:尿硫蝶呤、20-氧-白三烯E4、皮酮四醇和四氢可的松。
8.根据权利要求7所述的用途,其中监测髓母细胞瘤患者的预后是指监测髓母细胞瘤患者的术后病情恢复。
9.根据权利要求8所述的用途,其中如果髓母细胞瘤患者在术后尿液中代谢物水平具有以下变化,则所述髓母细胞瘤患者在术后恢复良好:尿硫蝶呤、20-氧-白三烯E4、皮酮四醇和四氢可的松中的一种或多种恢复至健康对照样本的水平,所述健康对照样本源自未患有髓母细胞瘤及其他疾病的健康个体的尿液。
10.一种用于早期诊断髓母细胞瘤和/或监测髓母细胞瘤患者预后的试剂盒或芯片,其包含检测以下代谢物水平的试剂:尿硫蝶呤、20-氧-白三烯E4和皮酮四醇;优选地,所述试剂盒或芯片用于质谱鉴定检测中。
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