Gene network information is believed to be beneficial for disease module and pathway identification, but has not been explicitly utilized in the standard random forest (RF) algorithm for gene expression data analysis. We investigate the performance of a network-guided RF where the network information is summarized into a sampling probability of predictor variables which is further used in the construction of the RF. Our results suggest that network-guided RF does not provide better disease prediction than the standard RF. In terms of disease gene discovery, if disease genes form module(s), network-guided RF identifies them more accurately. In addition, when disease status is independent from genes in the given network, spurious gene selection results can occur when using network information, especially on hub genes. Our empirical analysis on two balanced microarray and RNA-Seq breast cancer datasets from The Cancer Genome Atlas (TCGA) for classification of progesterone receptor (PR) status also demonstrates that network-guided RF can identify genes from PGR-related pathways, which leads to a better connected module of identified genes.

Deep learning based approaches have proven promising to model omics data. However, one of the current limitations compared to statistical and traditional machine learning approaches is the lack of explainability, which not only reduces the reliability, but limits the potential for acquiring novel knowledge from unpicking the "black-box" models. Here we present XOmiVAE, a novel interpretable deep learning model for cancer classification using high-dimensional omics data. XOmiVAE is able to obtain contribution values of each gene and latent dimension for a specific prediction, and the correlation between genes and the latent dimensions. It is also revealed that XOmiVAE can explain both the supervised classification and the unsupervised clustering results from the deep learning network. To the best of our knowledge, XOmiVAE is one of the first activated-based deep learning interpretation method to explain novel clusters generated by variational autoencoders. The results generated by XOmiVAE were validated by both the biomedical knowledge and the performance of downstream tasks. XOmiVAE explanations of deep learning based cancer classification and clustering aligned with current domain knowledge including biological annotation and literature, which shows great potential for novel biomedical knowledge discovery from deep learning models. The top XOmiVAE selected genes and dimensions shown significant influence to the performance of cancer classification. Additionally, we offer important steps to consider when interpreting deep learning models for tumour classification. For instance, we demonstrate the importance of choosing background samples that makes biological sense and the limitations of connection weight based methods to explain latent dimensions.

Over the past decades, statisticians and machine-learning researchers have developed literally thousands of new tools for the reduction of high-dimensional data in order to identify the variables most responsible for a particular trait. These tools have applications in a plethora of settings, including data analysis in the fields of business, education, forensics, and biology (such as microarray, proteomics, brain imaging), to name a few. In the present work, we focus our investigation on the limitations and potential misuses of certain tools in the analysis of the benchmark colon cancer data (2,000 variables; Alon et al., 1999) and the prostate cancer data (6,033 variables; Efron, 2010, 2008). Our analysis demonstrates that models that produce 100% accuracy measures often select different sets of genes and cannot stand the scrutiny of parameter estimates and model stability. Furthermore, we created a host of simulation datasets and "artificial diseases" to evaluate the reliability of commonly used statistical and data mining tools. We found that certain widely used models can classify the data with 100% accuracy without using any of the variables responsible for the disease. With moderate sample size and suitable pre-screening, stochastic gradient boosting will be shown to be a superior model for gene selection and variable screening from high-dimensional datasets.