一种用于评估肾移植活检的简单分子工具。
A Simple Molecular Tool for the Assessment of Kidney Transplant Biopsies.
机构信息
Nephrology - Kidney Transplant Unit, Rouen University Hospital, Rouen, France.
Univ Rouen Normandie, INSERM U1234, Rouen, France.
出版信息
Clin J Am Soc Nephrol. 2023 Apr 1;18(4):499-509. doi: 10.2215/CJN.0000000000000100. Epub 2023 Mar 8.
BACKGROUND
The Banff Classification for Allograft Pathology recommendations for the diagnosis of kidney transplant rejection includes molecular assessment of the transplant biopsy. However, implementation of molecular tools in clinical practice is still limited, partly due to the required expertise and financial investment. The reverse transcriptase multiplex ligation-dependent probe amplification (RT-MLPA) assay is a simple, rapid, and inexpensive assay that permits simultaneous evaluation of a restricted gene panel using paraffin-embedded tissue blocks. The aim of this study was to develop and validate a RT-MLPA assay for diagnosis and classification of rejection.
METHODS
A retrospective cohort of 220 kidney transplant biopsies from two centers, which included 52 antibody-mediated rejection, 51 T-cell-mediated rejection, and 117 no-rejection controls, was assessed. A 17-gene panel was identified on the basis of relevant pathophysiological pathways. A support vector machine classifier was developed. A subset of 109 biopsies was also assessed using the Nanostring Banff Human Organ Transplant panel to compare the two assays.
RESULTS
The support vector machine classifier train and test accuracy scores were 0.84 and 0.83, respectively. In the test cohort, the F1 score for antibody-mediated rejection, T-cell-mediated rejection, and control were 0.88, 0.86, and 0.69, respectively. Using receiver-operating characteristic curves, the area under the curve for class predictions was 0.96, 0.89, and 0.91, respectively, with a weighted average at 0.94. Classifiers' performances were highest for antibody-mediated rejection diagnosis with 94% correct predictions, compared with 88% correct predictions for control biopsies and 60% for T-cell-mediated rejection biopsies. Gene expression levels assessed by RT-MLPA and Nanostring were correlated: r = 0.68, P < 0.001. Equivalent gene expression profiles were obtained with both assays in 81% of the samples.
CONCLUSIONS
The 17-gene panel RT-MLPA assay, developed here for formalin-fixed paraffin-embedded kidney transplant biopsies, classified kidney transplant rejection with an overall accurate prediction ratio of 0.83.
PODCAST
This article contains a podcast at https://dts.podtrac.com/redirect.mp3/www.asn-online.org/media/podcast/CJASN/2023_04_10_CJN10100822.mp3.
背景
同种异体移植病理学的班夫分类建议用于诊断肾移植排斥反应，包括对移植活检进行分子评估。然而，分子工具在临床实践中的应用仍然有限，部分原因是需要专业知识和财务投资。逆转录多重连接依赖探针扩增（RT-MLPA）检测是一种简单、快速且廉价的检测方法，允许使用石蜡包埋组织块同时评估受限的基因组。本研究旨在开发和验证一种用于诊断和分类排斥反应的 RT-MLPA 检测方法。
方法
对来自两个中心的 220 例肾移植活检进行了回顾性队列研究，其中包括 52 例抗体介导的排斥反应、51 例 T 细胞介导的排斥反应和 117 例无排斥反应对照。基于相关病理生理途径确定了一个 17 基因的检测面板。开发了支持向量机分类器。还使用 Nanostring Banff 人类器官移植面板评估了 109 例活检，以比较两种检测方法。
结果
支持向量机分类器的训练和测试准确性评分分别为 0.84 和 0.83。在测试队列中，抗体介导的排斥反应、T 细胞介导的排斥反应和对照组的 F1 评分分别为 0.88、0.86 和 0.69。使用受试者工作特征曲线，分类预测的曲线下面积分别为 0.96、0.89 和 0.91，加权平均值为 0.94。抗体介导的排斥反应诊断的分类器性能最高，正确预测率为 94%，而对照活检的正确预测率为 88%，T 细胞介导的排斥反应活检的正确预测率为 60%。通过 RT-MLPA 和 Nanostring 评估的基因表达水平相关：r = 0.68，P < 0.001。两种检测方法在 81%的样本中获得了等效的基因表达谱。
结论
这里为福尔马林固定石蜡包埋的肾移植活检开发的 17 基因面板 RT-MLPA 检测方法，以 0.83 的总体准确预测率对肾移植排斥反应进行分类。
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