Quantitative immuno-mass spectrometry imaging of skeletal muscle dystrophin.
Quantitative immuno-mass spectrometry imaging of skeletal muscle dystrophin.
复制标题
骨骼肌肌营养不良蛋白的定量免疫质谱学成像。
DOI：
10.1038/s41598-020-80495-8
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发表时间：
2021-01-13
影响因子：
4.6
通讯作者：
Wanagat J
中科院分区：
文献类型：
--
作者：
Bishop DP;Westerhausen MT;Barthelemy F;Lockwood T;Cole N;Gibbs EM;Crosbie RH;Nelson SF;Miceli MC;Doble PA;Wanagat J
Duchenne肌营养不良症(DMD)的新兴和有希望的治疗干预措施因量化dystrophin的挑战而困惑。目前的方法精度低，需要大量的组织，而且很难标准化。本文提出了一种利用Gd标记的抗dystrophin抗体和激光消融-电感耦合等离子体质谱(LICMS)同时定量和定位肌肉切片中dystrophin的免疫质谱仪成像方法。Gd被量化为Dystrophin相对表达的替代指标，并在K-Means聚类分割后的小鼠和人类骨骼肌切片中得到验证，然后应用于具有不同基因突变的DMD患者，其中Dystrophin的表达被测得高达100ug/kg−1Gd。这些结果表明，免疫质谱仪成像是DMD临床前到临床研究的一种可行的方法。它可以快速定量单个组织切片中的相对肌营养不良蛋白，有效地利用宝贵的患者资源，并可能为临床翻译提供药物疗效的信息。
Emerging and promising therapeutic interventions for Duchenne muscular dystrophy (DMD) are confounded by the challenges of quantifying dystrophin. Current approaches have poor precision, require large amounts of tissue, and are difficult to standardize. This paper presents an immuno-mass spectrometry imaging method using gadolinium (Gd)-labeled anti-dystrophin antibodies and laser ablation-inductively coupled plasma-mass spectrometry to simultaneously quantify and localize dystrophin in muscle sections. Gd is quantified as a proxy for the relative expression of dystrophin and was validated in murine and human skeletal muscle sections following k-means clustering segmentation, before application to DMD patients with different gene mutations where dystrophin expression was measured up to 100 µg kg−1 Gd. These results demonstrate that immuno-mass spectrometry imaging is a viable approach for pre-clinical to clinical research in DMD. It rapidly quantified relative dystrophin in single tissue sections, efficiently used valuable patient resources, and may provide information on drug efficacy for clinical translation.