Contact: [email protected] Bioinformatics (2012) 28 (15), 1990–1997 doi:10.1093/bioinformatics/bts296 The authors wish to add one citation to a relevant conference report, Gu, J., Xuan, J., Wang, Y., Riggins, R.B. and Clarke R. (2010) Identification of transcriptional regulatory networks by learning the marginal function of outlier sum statistic. Proceedings of International Conference on Machine Learning and Applications , 281–286. The formatted reference is given below and should read in the sentence: In particular, a novel statistic for testing the confidence of target genes, namely, outlier sum of regression t -statistic ( Gu et al. , 2010 ), is specifically designed to pin-down confident target genes; based on this statistic, a Gibbs sampling strategy is used to sample target genes in a high probability as governed by the underlying distribution. The authors apologize for this oversight.

Reliable inference of transcription regulatory networks is a challenging task in computational biology. Network component analysis (NCA) has become a powerful scheme to uncover regulatory networks behind complex biological processes. However, the performance of NCA is impaired by the high rate of false connections in binding information. In this paper, we integrate stability analysis with NCA to form a novel scheme, namely stability-based NCA (sNCA), for regulatory network identification. The method mainly addresses the inconsistency between gene expression data and binding motif information. Small perturbations are introduced to prior regulatory network, and the distance among multiple estimated transcript factor (TF) activities is computed to reflect the stability for each TF's binding network. For target gene identification, multivariate regression and t-statistic are used to calculate the significance for each TF-gene connection. Simulation studies are conducted and the experimental results show that sNCA can achieve an improved and robust performance in TF identification as compared to NCA. The approach for target gene identification is also demonstrated to be suitable for identifying true connections between TFs and their target genes. Furthermore, we have successfully applied sNCA to breast cancer data to uncover the role of TFs in regulating endocrine resistance in breast cancer.

MOTIVATION: Identification of transcriptional regulatory networks (TRNs) is of significant importance in computational biology for cancer research, providing a critical building block to unravel disease pathways. However, existing methods for TRN identification suffer from the inclusion of excessive 'noise' in microarray data and false-positives in binding data, especially when applied to human tumor-derived cell line studies. More robust methods that can counteract the imperfection of data sources are therefore needed for reliable identification of TRNs in this context. RESULTS: In this article, we propose to establish a link between the quality of one target gene to represent its regulator and the uncertainty of its expression to represent other target genes. Specifically, an outlier sum statistic was used to measure the aggregated evidence for regulation events between target genes and their corresponding transcription factors. A Gibbs sampling method was then developed to estimate the marginal distribution of the outlier sum statistic, hence, to uncover underlying regulatory relationships. To evaluate the effectiveness of our proposed method, we compared its performance with that of an existing sampling-based method using both simulation data and yeast cell cycle data. The experimental results show that our method consistently outperforms the competing method in different settings of signal-to-noise ratio and network topology, indicating its robustness for biological applications. Finally, we applied our method to breast cancer cell line data and demonstrated its ability to extract biologically meaningful regulatory modules related to estrogen signaling and action in breast cancer. AVAILABILITY AND IMPLEMENTATION: The Gibbs sampler MATLAB package is freely available at http://www.cbil.ece.vt.edu/software.htm. CONTACT: [email protected] SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

BACKGROUND: Genes work coordinately as gene modules or gene networks. Various computational approaches have been proposed to find gene modules based on gene expression data; for example, gene clustering is a popular method for grouping genes with similar gene expression patterns. However, traditional gene clustering often yields unsatisfactory results for regulatory module identification because the resulting gene clusters are co-expressed but not necessarily co-regulated. RESULTS: We propose a novel approach, motif-guided sparse decomposition (mSD), to identify gene regulatory modules by integrating gene expression data and DNA sequence motif information. The mSD approach is implemented as a two-step algorithm comprising estimates of (1) transcription factor activity and (2) the strength of the predicted gene regulation event(s). Specifically, a motif-guided clustering method is first developed to estimate the transcription factor activity of a gene module; sparse component analysis is then applied to estimate the regulation strength, and so predict the target genes of the transcription factors. The mSD approach was first tested for its improved performance in finding regulatory modules using simulated and real yeast data, revealing functionally distinct gene modules enriched with biologically validated transcription factors. We then demonstrated the efficacy of the mSD approach on breast cancer cell line data and uncovered several important gene regulatory modules related to endocrine therapy of breast cancer. CONCLUSION: We have developed a new integrated strategy, namely motif-guided sparse decomposition (mSD) of gene expression data, for regulatory module identification. The mSD method features a novel motif-guided clustering method for transcription factor activity estimation by finding a balance between co-regulation and co-expression. The mSD method further utilizes a sparse decomposition method for regulation strength estimation. The experimental results show that such a motif-guided strategy can provide context-specific regulatory modules in both yeast and breast cancer studies.

Network component analysis (NCA) and other methods based on the NCA model have become powerful bioinformatics tools to reconstruct underlying regulatory networks and recover hidden biological processes. However, due to the existence of experimental noises in micro array data and false information in network connectivity data (e.g., ChIP-on-chip binding data, motif information, etc.), it still remains challenging to reconstruct gene regulatory networks for real biomedical applications such as human cancer studies. In this paper, we model the relationship between the genes that share the same transcription factors (TF) from the angle of regression. We propose a statistic called outlier sum testing the conditional significance of the target genes. A Gibbs strategy is utilized in order to estimate the marginal value of outlier sum from its conditional function. Based on the outlier sum statistic we are able to extract the true target genes that carry information about transcription factor activities (TFAs) from the whole population. As a proof-of-concept, we demonstrated the efficiency and robustness of the proposed method on both simulation data and yeast cell cycle data.

MOTIVATION: The identification of gene regulatory modules is an important yet challenging problem in computational biology. While many computational methods have been proposed to identify regulatory modules, their initial success is largely compromised by a high rate of false positives, especially when applied to human cancer studies. New strategies are needed for reliable regulatory module identification. RESULTS: We present a new approach, namely multilevel support vector regression (ml-SVR), to systematically identify condition-specific regulatory modules. The approach is built upon a multilevel analysis strategy designed for suppressing false positive predictions. With this strategy, a regulatory module becomes ever more significant as more relevant gene sets are formed at finer levels. At each level, a two-stage support vector regression (SVR) method is utilized to help reduce false positive predictions by integrating binding motif information and gene expression data; a significant analysis procedure is followed to assess the significance of each regulatory module. To evaluate the effectiveness of the proposed strategy, we first compared the ml-SVR approach with other existing methods on simulation data and yeast cell cycle data. The resulting performance shows that the ml-SVR approach outperforms other methods in the identification of both regulators and their target genes. We then applied our method to breast cancer cell line data to identify condition-specific regulatory modules associated with estrogen treatment. Experimental results show that our method can identify biologically meaningful regulatory modules related to estrogen signaling and action in breast cancer. AVAILABILITY AND IMPLEMENTATION: The ml-SVR MATLAB package can be downloaded at http://www.cbil.ece.vt.edu/software.htm.

MOTIVATION: Significant efforts have been made to acquire data under different conditions and to construct static networks that can explain various gene regulation mechanisms. However, gene regulatory networks are dynamic and condition-specific; under different conditions, networks exhibit different regulation patterns accompanied by different transcriptional network topologies. Thus, an investigation on the topological changes in transcriptional networks can facilitate the understanding of cell development or provide novel insights into the pathophysiology of certain diseases, and help identify the key genetic players that could serve as biomarkers or drug targets. RESULTS: Here, we report a differential dependency network (DDN) analysis to detect statistically significant topological changes in the transcriptional networks between two biological conditions. We propose a local dependency model to represent the local structures of a network by a set of conditional probabilities. We develop an efficient learning algorithm to learn the local dependency model using the Lasso technique. A permutation test is subsequently performed to estimate the statistical significance of each learned local structure. In testing on a simulation dataset, the proposed algorithm accurately detected all the genes with network topological changes. The method was then applied to the estrogen-dependent T-47D estrogen receptor-positive (ER+) breast cancer cell line datasets and human and mouse embryonic stem cell datasets. In both experiments using real microarray datasets, the proposed method produced biologically meaningful results. We expect DDN to emerge as an important bioinformatics tool in transcriptional network analyses. While we focus specifically on transcriptional networks, the DDN method we introduce here is generally applicable to other biological networks with similar characteristics. AVAILABILITY: The DDN MATLAB toolbox and experiment data are available at http://www.cbil.ece.vt.edu/software.htm.

One of the major goals in microarray data analysis is to identify biomarkers and build a classification model for future prediction. Many traditional statistical models, based on microarray data alone, often fail in identifying biologically meaningful genes, which should have synergistic effect on determine the clinical outcomes through some interactions rather than work individually. In this paper, we proposed a network-constrained support vector machine (nSVM) for classification by incorporating prior knowledge, which could be protein-protein interactions, protein-gene regulation relationships or pathways information. Specifically, we use Laplacian matrix to represent gene-gene interaction network to regularize the objective function of SVM, which imposes the smoothness of coefficients over the network. The experimental results on simulation and real microarray datasets demonstrate that our method could not only improve classification performance compared to conventional SVM, but more importantly, it could identify significant sub-networks belonging to several pathways which might be related to underlying mechanism associated with clinical outcomes.