Transcriptional Regulatory Networks across Species:Evolution, Inference, and Refinement

The determination of transcriptional regulatory networks is key to the understanding of biological systems. However, the experimental determination of transcriptional regulatory networks in the laboratory remains difficult and time-consuming, while current computational methods to infer these networks (typically from gene-expression data) achieve only modest accuracy. The latter can be attributed in part to the limitations of a single-organism approach. Computational biology has long used comparative and, more generally, evolutionary approaches to extend the reach and accuracy of its analyses. We therefore use an evolutionary approach to the inference of regulatory networks, which enables us to study evolutionary models for these networks as well as to improve the accuracy of inferred networks. Since the regulatory networks evolve along with the genomes, we consider that the regulatory networks for a family of organisms are related to each other through the same phylogenetic tree. These relationships contain information that can be used to improve the accuracy of inferred networks. Advances in the study of evolution of regulatory networks provide evidence to establish evolutionary models for regulatory networks, which is an important component of our evolutionary approach. We use two network evolutionary models, a basic model that considers only the gains and losses of regulatory connections during evolution, and an extended model that also takes into account the duplications and losses of genes. With the network evolutionary models, we design refinement algorithms to make use of the phylogenetic relationships to refine noisy regulatory networks for a family of organisms. These refinement algorithms include: RefineFast and RefineML, which are two-step iterative algorithms, and ProPhyC and ProPhyCC, which are based on a probabilistic phylogenetic model. For each algorithm we first design it with the basic network evolutionary model and then generalize it to the extended evolutionary model. All these algorithms are computationally efficient and are supported by extensive experimental results showing that they yield substantial improvement in the quality of the input noisy networks. In particular, ProPhyC and ProPhyCC further improve the performance of RefineFast and RefineML. Besides the four refinement algorithms mentioned above, we also design an algorithm based on transfer learning theory called tree transfer learning (TTL). TTL differs from the previous four refinement algorithms in the sense that it takes the gene-expression data for the family of organisms as input, instead of their inferred noisy networks. TTL then learns the network structures for all the organisms at once, meanwhile taking advantage of the phylogenetic relationships. Although this approach outperforms an inference algorithm used alone, it does not perform better than ProPhyC, which indicates that the ProPhyC framework makes good use of the phylogenetic information

Development of Computational Techniques for Regulatory DNA Motif Identification Based on Big Biological Data

Accurate regulatory DNA motif (or motif) identification plays a fundamental role in the elucidation of transcriptional regulatory mechanisms in a cell and can strongly support the regulatory network construction for both prokaryotic and eukaryotic organisms. Next-generation sequencing techniques generate a huge amount of biological data for motif identification. Specifically, Chromatin Immunoprecipitation followed by high throughput DNA sequencing (ChIP-seq) enables researchers to identify motifs on a genome scale. Recently, technological improvements have allowed for DNA structural information to be obtained in a high-throughput manner, which can provide four DNA shape features. The DNA shape has been found as a complementary factor to genomic sequences in terms of transcription factor (TF)-DNA binding specificity prediction based on traditional machine learning models. Recent studies have demonstrated that deep learning (DL), especially the convolutional neural network (CNN), enables identification of motifs from DNA sequence directly. Although numerous algorithms and tools have been proposed and developed in this field, (1) the lack of intuitive and integrative web servers impedes the progress of making effective use of emerging algorithms and tools; (2) DNA shape has not been integrated with DL; and (3) existing DL models still suffer high false positive and false negative issues in motif identification. This thesis focuses on developing an integrated web server for motif identification based on DNA sequences either from users or built-in databases. This web server allows further motif-related analysis and Cytoscape-like network interpretation and visualization. We then proposed a DL framework for both sequence and shape motif identification from ChIP-seq data using a binomial distribution strategy. This framework can accept as input the different combinations of DNA sequence and DNA shape. Finally, we developed a gated convolutional neural network (GCNN) for capturing motif dependencies among long DNA sequences. Results show that our developed web server enables providing comprehensive motif analysis functionalities compared with existing web servers. The DL framework can identify motifs using an optimized threshold and disclose the strong predictive power of DNA shape in TF-DNA binding specificity. The identified sequence and shape motifs can contribute to TF-DNA binding mechanism interpretation. Additionally, GCNN can improve TF-DNA binding specificity prediction than CNN on most of the datasets

In silico pathway reconstruction: Iron-sulfur cluster biogenesis in Saccharomyces cerevisiae

BACKGROUND: Current advances in genomics, proteomics and other areas of molecular biology make the identification and reconstruction of novel pathways an emerging area of great interest. One such class of pathways is involved in the biogenesis of Iron-Sulfur Clusters (ISC). RESULTS: Our goal is the development of a new approach based on the use and combination of mathematical, theoretical and computational methods to identify the topology of a target network. In this approach, mathematical models play a central role for the evaluation of the alternative network structures that arise from literature data-mining, phylogenetic profiling, structural methods, and human curation. As a test case, we reconstruct the topology of the reaction and regulatory network for the mitochondrial ISC biogenesis pathway in S. cerevisiae. Predictions regarding how proteins act in ISC biogenesis are validated by comparison with published experimental results. For example, the predicted role of Arh1 and Yah1 and some of the interactions we predict for Grx5 both matches experimental evidence. A putative role for frataxin in directly regulating mitochondrial iron import is discarded from our analysis, which agrees with also published experimental results. Additionally, we propose a number of experiments for testing other predictions and further improve the identification of the network structure. CONCLUSION: We propose and apply an iterative in silico procedure for predictive reconstruction of the network topology of metabolic pathways. The procedure combines structural bioinformatics tools and mathematical modeling techniques that allow the reconstruction of biochemical networks. Using the Iron Sulfur cluster biogenesis in S. cerevisiae as a test case we indicate how this procedure can be used to analyze and validate the network model against experimental results. Critical evaluation of the obtained results through this procedure allows devising new wet lab experiments to confirm its predictions or provide alternative explanations for further improving the models

Analysis of minimal metabolic networks through whole-cell in silico modelling of prokaryotes

Tese de Mestrado Integrado. Bioengenharia. Faculdade de Engenharia. Universidade do Porto. 201

Modular Algorithms for Biomolecular Network Alignment

Comparative analysis of biomolecular networks constructed using measurements from different conditions, tissues, and organisms offer a powerful approach to understanding the structure, function, dynamics, and evolution of complex biological systems. The rapidly advancing field of systems biology aims to understand the structure, function, dynamics, and evolution of complex biological systems in terms of the underlying networks of interactions among the large number of molecular participants involved including genes, proteins, and metabolites. In particular, the comparative analysis of network models representing biomolecular interactions in different species or tissues offers an important tool for identifying conserved modules, predicting functions of specific genes or proteins and studying the evolution of biological processes, among other applications. The primary focus of this dissertation is on the biomolecular network alignment problem: Given two or more network models, the problem is to optimally match the nodes and links in one network with the nodes and links of the other. The Biomolecular Network Alignment (BiNA) Toolkit developed as part of this dissertation provides a set of efficient (in terms of the running time complexity) and accurate (in terms of various evaluation criteria discussed in the literature) network alignment algorithms for biomolecular networks. BiNA is scalable, user-friendly, modular, and extensible for performing alignments on diverse types of biomolecular networks. The algorithm is applicable to (1) undirected graphs in their weighted and unweighted variations (2) undirected graphs in their labeled and unlabeled variations (3) and has been applied to align multiple networks from hundreds of nodes with a few thousand edges to networks with tens of thousands of nodes with millions of edges. The dissertation provides various applications of network comparison tools including how results from such alignments have been utilized to (1) construct phylogenetic trees based on protein-protein interaction networks, and (2) find biochemical pathways involved in ligand recognition in B cells

Eric Davidson: Steps to A Gene Regulatory Network for Development

Eric Harris Davidson was a unique and creative intellectual force who grappled with the diversity of developmental processes used by animal embryos and wrestled them into an intelligible set of principles, then spent his life translating these process elements into molecularly definable terms through the architecture of gene regulatory networks. He took speculative risks in his theoretical writing but ran a highly organized, rigorous experimental program that yielded an unprecedentedly full characterization of a developing organism. His writings created logical order and a framework for mechanism from the complex phenomena at the heart of advanced multicellular organism development. This is a reminiscence of intellectual currents in his work as observed by the author through the last 30-35 years of Davidson's life

Inferring interaction networks from transcriptomic data: methods and applications

Transcriptomic data is a treasure-trove in modern molecular biology, as it offers a comprehensive viewpoint into the intricate nuances of gene expression dynamics underlying biological systems. This genetic information must be utilised to infer biomolecular interaction networks that can provide insights into the complex regulatory mechanisms underpinning the dynamic cellular processes. Gene regulatory networks and protein-protein interaction networks are two major classes of such networks. This chapter thoroughly investigates the wide range of methodologies used for distilling insightful revelations from transcriptomic data that include association based methods (based on correlation among expression vectors), probabilistic models (using Bayesian and Gaussian models), and interologous methods. We reviewed different approaches for evaluating the significance of interactions based on the network topology and biological functions of the interacting molecules, and discuss various strategies for the identification of functional modules. The chapter concludes with highlighting network based techniques of prioritising key genes, outlining the centrality based, diffusion based and subgraph based methods. The chapter provides a meticulous framework for investigating transcriptomic data to uncover assembly of complex molecular networks for their adaptable analyses across a broad spectrum of biological domains.Comment: 48 pages, 3 figure