K.G. Jebsen Coeliac Disease Research Centre, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
K.G. Jebsen Coeliac Disease Research Centre, Institute of Clinical Medicine, University of Oslo, Oslo, Norway; Department of Informatics, The Faculty of Mathematics and Natural Sciences, University of Oslo, Oslo, Norway.
Gastroenterology. 2024 Aug;167(3):493-504.e10. doi: 10.1053/j.gastro.2024.03.006. Epub 2024 Mar 11.
BACKGROUND & AIMS: Histologic evaluation of gut biopsies is a cornerstone for diagnosis and management of celiac disease (CeD). Despite its wide use, the method depends on proper biopsy orientation, and it suffers from interobserver variability. Biopsy proteome measurement reporting on the tissue state can be obtained by mass spectrometry analysis of formalin-fixed paraffin-embedded tissue. Here we aimed to transform biopsy proteome data into numerical scores that give observer-independent measures of mucosal remodeling in CeD.
A pipeline using glass-mounted formalin-fixed paraffin-embedded sections for mass spectrometry-based proteome analysis was established. Proteome data were converted to numerical scores using 2 complementary approaches: a rank-based enrichment score and a score based on machine learning using logistic regression. The 2 scoring approaches were compared with each other and with histology analyzing 18 patients with CeD with biopsies collected before and after treatment with a gluten-free diet as well as biopsies from patients with CeD with varying degree of remission (n = 22). Biopsies from individuals without CeD (n = 32) were also analyzed.
The method yielded reliable proteome scoring of both unstained and H&E-stained glass-mounted sections. The scores of the 2 approaches were highly correlated, reflecting that both approaches pick up proteome changes in the same biological pathways. The proteome scores correlated with villus height-to-crypt depth ratio. Thus, the method is able to score biopsies with poor orientation.
Biopsy proteome scores give reliable observer and orientation-independent measures of mucosal remodeling in CeD. The proteomic method can readily be implemented by nonexpert laboratories in parallel to histology assessment and easily scaled for clinical trial settings.
对肠道活检进行组织学评估是诊断和治疗乳糜泻(CeD)的基石。尽管这种方法应用广泛,但它依赖于适当的活检方向,并且存在观察者间的变异性。通过对福尔马林固定石蜡包埋组织进行质谱分析,可以获得活检蛋白质组测量对组织状态的报告。在这里,我们旨在将活检蛋白质组数据转化为数字评分,为 CeD 提供观察者独立的黏膜重塑测量。
建立了一种使用载玻片固定的福尔马林石蜡包埋切片进行基于质谱的蛋白质组分析的流水线。使用 2 种互补方法将蛋白质组数据转换为数字评分:基于排名的富集评分和基于逻辑回归的机器学习评分。这 2 种评分方法相互比较,并与分析 18 例 CeD 患者的活检进行比较,这些患者在接受无麸质饮食治疗前后采集了活检,以及分析不同缓解程度的 CeD 患者的活检(n=22)。还分析了无 CeD 个体的活检(n=32)。
该方法可靠地对未经染色和 H&E 染色的载玻片切片进行蛋白质组评分。这 2 种方法的评分高度相关,反映出这 2 种方法都能检测到相同生物学途径中的蛋白质组变化。蛋白质组评分与绒毛高度与隐窝深度比相关。因此,该方法能够对定向不良的活检进行评分。
活检蛋白质组评分可提供 CeD 黏膜重塑的可靠观察者和定向独立测量。非专业实验室可以很容易地将蛋白质组方法与组织学评估并行实施,并容易扩展到临床试验设置。