Tandem Mass Tag-Based Serum Proteome Profiling for Biomarker Discovery in Young Duchenne Muscular Dystrophy Boys.
Tandem Mass Tag-Based Serum Proteome Profiling for Biomarker Discovery in Young Duchenne Muscular Dystrophy Boys.
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DOI：
10.1021/acsomega.0c03206
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发表时间：
2020-10-20
期刊：
影响因子：
4.1
通讯作者：
Hathout Y
中科院分区：
文献类型：
--
作者：
Alayi TD;Tawalbeh SM;Ogundele M;Smith HR;Samsel AM;Barbieri ML;Hathout Y
血液可及分子生物标记物正成为评估Duchenne肌营养不良症(DMD)疾病进展和治疗反应的极具吸引力的工具，特别是在非常年轻的患者中，对他们来说，其他结果指标仍然是主观的和具有挑战性的。在这项研究中，我们标准化了一种高度特异和可重复性的多重质谱学方法，使用串联质量标签(TMT)策略，结合从血清中去除丰富的蛋白质和高pH反相多肽分级。4岁DMD男童(n=9)和年龄匹配的健康对照组(n=9)的差异蛋白质组图谱显示，与健康对照组相比，DMD组有38个升高的和50个降低的血清蛋白质(调整后P<；0.05，FDR<；0.05)。正如预期的那样，我们确认了之前报道的生物标记物，但也发现了新的生物标记物。这些包括新的肌肉损伤相关生物标记物，如端乙素、平滑蛋白样蛋白1、Cofilin-1和凝集素，其他肌肉特异性酶，如UTP-葡萄糖-1-磷酸尿苷转移酶、天冬氨酸氨基转移酶、丙酮酸激酶PKM、乳转铁蛋白、组织α-L岩藻糖苷酶、泛氨酸酶和非球蛋白-1，以及一些促炎和细胞黏附相关生物标记物，如白涎素、巨噬细胞受体MARCO、Vitronectin、Galectin-3结合蛋白和ProSAAS。包括血清去除、样品处理和质谱分析在内的工作流程被发现是重复性的，随着时间的推移，变异系数为20%。此外，与其他基于多重亲和力的方法相比，该方法被发现在特异性方面更好。这些发现证明了基于TMT的质谱学方法在检测和鉴定无症状的年轻DMD患者的血清生物标记物方面的特异性和可靠性。
Blood-accessible molecular biomarkers are becoming highly attractive tools to assess disease progression and response to therapies in Duchenne muscular dystrophy (DMD) especially in very young patients for whom other outcome measures remain subjective and challenging. In this study, we have standardized a highly specific and reproducible multiplexing mass spectrometry method using the tandem mass tag (TMT) strategy in combination with depletion of abundant proteins from serum and high-pH reversed-phase peptide fractionation. Differential proteome profiling of 4 year-old DMD boys (n = 9) and age-matched healthy controls (n = 9) identified 38 elevated and 50 decreased serum proteins (adjusted P < 0.05, FDR <0.05) in the DMD group relative to the healthy control group. As expected, we confirmed previously reported biomarkers but also identified novel biomarkers. These included novel muscle injury-associated biomarkers such as telethonin, smoothelin-like protein 1, cofilin-1, and plectin, additional muscle-specific enzymes such as UTP–glucose-1-phosphate uridylyltransferase, aspartate aminotransferase, pyruvate kinase PKM, lactotransferrin, tissue alpha-l-fucosidase, pantetheinase, and ficolin-1, and some pro-inflammatory and cell adhesion-associated biomarkers such as leukosialin, macrophage receptor MARCO, vitronectin, galectin-3-binding protein, and ProSAAS. The workflow including serum depletion, sample processing, and mass spectrometry analysis was found to be reproducible and stable over time with CV < 20%. Furthermore, the method was found to be superior in terms of specificity compared to other multiplexing affinity-based methods. These findings demonstrate the specificity and reliability of TMT-based mass spectrometry methods in detection and identification of serum biomarkers in presymptomatic young DMD patients.
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DOI：
10.1155/2013/219452
发表时间：
2013
期刊：
International journal of proteomics
影响因子：
--
作者：
Hale JE
通讯作者：
Hale JE
影响因子：
3.4
作者：
Mcdonald, Craig M.;Henricson, Erik K.;Abresch, R. Ted;Florence, Julaine M.;Eagle, Michelle;Gappmaier, Eduard;Glanzman, Allan M.;Spiegel, Robert;Barth, Jay;Elfring, Gary;Reha, Allen;Peltz, Stuart
通讯作者：
Peltz, Stuart
影响因子：
11.2
作者：
Mendell, Jerry R.;Shilling, Chris;Weiss, Robert B.
通讯作者：
Weiss, Robert B.
影响因子：
3.3
作者：
Khoonsari, Payam Emami;Musunri, Sravani;Kultima, Kim
通讯作者：
Kultima, Kim
DOI：
10.1073/pnas.1507719112
发表时间：
2015-06-09
影响因子：
11.1
作者：
Hathout Y;Brody E;Clemens PR;Cripe L;DeLisle RK;Furlong P;Gordish-Dressman H;Hache L;Henricson E;Hoffman EP;Kobayashi YM;Lorts A;Mah JK;McDonald C;Mehler B;Nelson S;Nikrad M;Singer B;Steele F;Sterling D;Sweeney HL;Williams S;Gold L
通讯作者：
Gold L