MOTIVATION: Correctly identifying epitope-binding T-cell receptors (TCRs) is important to both understand their underlying biological mechanism in association to some phenotype and accordingly develop T-cell mediated immunotherapy treatments. Although the importance of the CDR3 region in TCRs for epitope recognition is well recognized, methods for profiling their interactions in association to a certain disease or phenotype remains less studied. We developed EpicPred to identify phenotype-specific TCR-epitope interactions. EpicPred first predicts and removes unlikely TCR-epitope interactions to reduce false positives using the Open-set Recognition (OSR). Subsequently, multiple instance learning was used to identify TCR-epitope interactions specific to a cancer type or severity levels of COVID-19 infected patients. RESULTS: From six public TCR databases, 244 552 TCR sequences and 105 unique epitopes were used to predict epitope-binding TCRs and to filter out non-epitope-binding TCRs using the OSR method. The predicted interactions were used to further predict the phenotype groups in two cancer and four COVID-19 TCR-seq datasets of both bulk and single-cell resolution. EpicPred outperformed the competing methods in predicting the phenotypes, achieving an average AUROC of 0.80 ± 0.07. AVAILABILITY AND IMPLEMENTATION: The EpicPred Software is available at https://github.com/jaeminjj/EpicPred.

BACKGROUND: Selecting informative genes or eliminating uninformative ones before any downstream gene expression analysis is a standard task with great impact on the results. A carefully curated gene set significantly enhances the likelihood of identifying meaningful biomarkers. METHOD: In contrast to the conventional forward gene search methods that focus on selecting highly informative genes, we propose a backward search method, DenoiseIt, that aims to remove potential outlier genes yielding a robust gene set with reduced noise. The gene set constructed by DenoiseIt is expected to capture biologically significant genes while pruning irrelevant ones to the greatest extent possible. Therefore, it also enhances the quality of downstream comparative gene expression analysis. DenoiseIt utilizes non-negative matrix factorization in conjunction with isolation forests to identify outlier rank features and remove their associated genes. RESULTS: DenoiseIt was applied to both bulk and single-cell RNA-seq data collected from TCGA and a COVID-19 cohort to show that it proficiently identified and removed genes exhibiting expression anomalies confined to specific samples rather than a known group. DenoiseIt also showed to reduce the level of technical noise while preserving a higher proportion of biologically relevant genes compared to existing methods. The DenoiseIt Software is publicly available on GitHub at https://github.com/cobi-git/DenoiseIt.

Recent advances in single-cell RNA sequencing (scRNA-seq) technology have enabled the acquisition of RNA at the single-cell level, which showed that the expression level of genes is highly variable across and within the cell types. Even well-known housekeeping genes showed high expression variance in a single condition and within the same cell types. Previous studies made efforts to identify stably expressed genes and use them as a yardstick for robust gene expression normalization. On the other hand, drugs were shown to be less effective on genes with high expression variance. Thus, identifying both stably and variably expressed genes is an important task, especially at the single-cell level. In this study, using the Kullback-Leibler divergence method, we proposed a metric to measure the expression stability of each gene. Using private scRNA-seq data composed of 25 severe COVID-19 patients and 40 healthy individuals, we identified variably expressed genes specific to COVID-19-infected patients and healthy cohorts.