Diagnosis of T-cell-mediated kidney rejection by biopsy-based proteomic biomarkers and machine learning

Abstract

BackgroundBiopsy-based diagnosis is essential for maintaining kidney allograft longevity by ensuring prompt treatment for graft complications. Although histologic assessment remains the gold standard, it carries significant limitations such as subjective interpretation, suboptimal reproducibility, and imprecise quantitation of disease burden. It is hoped that molecular diagnostics could enhance the efficiency, accuracy, and reproducibility of traditional histologic methods.MethodsQuantitative label-free mass spectrometry analysis was performed on a set of formalin-fixed, paraffin-embedded (FFPE) biopsies from kidney transplant patients, including five samples each with diagnosis of T-cell-mediated rejection (TCMR), polyomavirus BK nephropathy (BKPyVN), and stable (STA) kidney function control tissue. Using the differential protein expression result as a classifier, three different machine learning algorithms were tested to build a molecular diagnostic model for TCMR.ResultsThe label-free proteomics method yielded 800-1350 proteins that could be quantified with high confidence per sample by single-shot measurements. Among these candidate proteins, 329 and 467 proteins were defined as differentially expressed proteins (DEPs) for TCMR in comparison with STA and BKPyVN, respectively. Comparing the FFPE quantitative proteomics data set obtained in this study using label-free method with a data set we previously reported using isobaric labeling technology, a classifier pool comprised of features from DEPs commonly quantified in both data sets, was generated for TCMR prediction. Leave-one-out cross-validation result demonstrated that the random forest (RF)-based model achieved the best predictive power. In a follow-up blind test using an independent sample set, the RF-based model yields 80% accuracy for TCMR and 100% for STA. When applying the established RF-based model to two public transcriptome datasets, 78.1%-82.9% sensitivity and 58.7%-64.4% specificity was achieved respectively.ConclusionsThis proof-of-principle study demonstrates the clinical feasibility of proteomics profiling for FFPE biopsies using an accurate, efficient, and cost-effective platform integrated of quantitative label-free mass spectrometry analysis with a machine learning-based diagnostic model. It costs less than 10 dollars per test.

Revolutionizing Kidney Transplantation: Connecting Machine Learning and Artificial Intelligence with Next-Generation Healthcare—From Algorithms to Allografts

“…Fran et al [79] employed a quantitative label-free mass spectrometry analysis on biopsies to develop a molecular diagnostic model for T-cell-mediated rejection (TCMR). Their random forest (RF) model achieved an accuracy of 0.80 for TCMR detection and 100% accuracy for stable kidney function in blind tests.…”

The transformative potential of artificial intelligence in solid organ transplantation

“…Efforts have been made recently to use AI to integrate transcriptomic or proteomic data to establish diagnostic models ( 27 ). For instance, Fang et al used quantitative label-free mass spectrometry analysis on formalin-fixed, paraffin-embedded biopsies from kidney transplant patients to identify proteomic biomarkers for T-cell-mediated kidney rejection ( 28 ). The label-free proteomics method allowed for quantifying 800–1,350 proteins per sample with high confidence.…”