Kruijt Mirjam, de la Morena-Barrio Maria Eugenia, Corral Javier, Cobbaert Christa M, Ruhaak L Renee
Department of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center, Leiden, the Netherlands.
Servicio de Hematología, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, CIBERER-ISCIII, Murcia, Spain.
J Thromb Haemost. 2025 Jan;23(1):210-221. doi: 10.1016/j.jtha.2024.10.005. Epub 2024 Oct 17.
Although P5 (preventive, personalized, predictive, participatory, psychocognitive) medicine and patient-focused healthcare are gaining ground in various healthcare areas, the diagnosis of antithrombin deficiency (ATD) is still based on crude diagnostic tests, clustering patients into clinically heterogeneous subgroups whereby relevant thrombophilia phenotypes may go unnoticed. Clinical pathways and the majority of evidence are based on these tests; therefore, generic treatment is still the norm.
To unravel the heterogeneity of ATD, a mass spectrometry (liquid chromatography coupled to multiple-reaction-monitoring mass spectrometry [LC-MRM-MS])-based test for antithrombin was developed allowing molecular characterization of the antithrombin proteoforms in patient plasma. This study provides the first insight into the tests' clinical performance.
Plasma from 91 unrelated ATD patients and 41 patients with a congenital disorder of glycosylation affecting antithrombin glycosylation were characterized functionally, genetically, and analyzed by LC-MRM-MS. An established data analysis strategy was applied for quantitation and molecular characterization of antithrombin proteoforms.
The test recognized patients with a quantitative defect, discriminated between type I and type II ATD, and identified variant proteoforms. Overall, the diagnostic sensitivity for ATD was 100% for LC-MRM-MS compared with 81.1% by the functional test. Type II ATD, a subtype prone to misdiagnosis, revealed an even larger difference of 100% identification by LC-MRM-MS vs 56.8% by functional test.
The qualitative and quantitative mass spectrometry-based AT-test can serve as a platform for investigating the molecular basis of the clinical heterogeneity of ATD. This "precision diagnostics" approach for ATD can lower diagnostic uncertainty and modernize the ATD diagnostic and clinical pathways.
尽管P5（预防、个性化、预测、参与、心理认知）医学和以患者为中心的医疗保健在各个医疗领域逐渐兴起，但抗凝血酶缺乏症（ATD）的诊断仍基于粗略的诊断测试，将患者归为临床异质性亚组，从而可能忽略相关的血栓形成倾向表型。临床路径和大多数证据都基于这些测试；因此，通用治疗仍是常态。
为了解开ATD的异质性，开发了一种基于质谱（液相色谱-多反应监测质谱联用[LC-MRM-MS]）的抗凝血酶测试，可对患者血浆中的抗凝血酶蛋白亚型进行分子表征。本研究首次深入了解了该测试的临床性能。
对91例无亲缘关系的ATD患者和41例患有影响抗凝血酶糖基化的先天性糖基化障碍患者的血浆进行功能、基因特征分析，并通过LC-MRM-MS进行分析。采用既定的数据分析策略对抗凝血酶蛋白亚型进行定量和分子表征。
该测试能够识别出存在定量缺陷的患者，区分I型和II型ATD，并鉴定出变异蛋白亚型。总体而言，LC-MRM-MS对ATD的诊断敏感性为100%，而功能测试为81.1%。II型ATD是一种容易误诊的亚型，LC-MRM-MS的识别率为100%，而功能测试为56.8%，两者差异更大。
基于质谱的定性和定量抗凝血酶测试可作为研究ATD临床异质性分子基础的平台。这种针对ATD的“精准诊断”方法可降低诊断不确定性，并使ATD诊断和临床路径现代化。
