Interrogating the topological robustness of gene regulatory circuits by randomization

Abstract

One of the most important roles of cells is performing their cellular tasks properly for survival. Cells usually achieve robust functionality, for example, cell-fate decision-making and signal transduction, through multiple layers of regulation involving many genes. Despite the combinatorial complexity of gene regulation, its quantitative behavior has been typically studied on the basis of experimentally verified core gene regulatory circuitry, composed of a small set of important elements. It is still unclear how such a core circuit operates in the presence of many other regulatory molecules and in a crowded and noisy cellular environment. Here we report a new computational method, named random circuit perturbation (RACIPE), for interrogating the robust dynamical behavior of a gene regulatory circuit even without accurate measurements of circuit kinetic parameters. RACIPE generates an ensemble of random kinetic models corresponding to a fixed circuit topology, and utilizes statistical tools to identify generic properties of the circuit. By applying RACIPE to simple toggle-switch-like motifs, we observed that the stable states of all models converge to experimentally observed gene state clusters even when the parameters are strongly perturbed. RACIPE was further applied to a proposed 22-gene network of the Epithelial-to-Mesenchymal Transition (EMT), from which we identified four experimentally observed gene states, including the states that are associated with two different types of hybrid Epithelial/Mesenchymal phenotypes. Our results suggest that dynamics of a gene circuit is mainly determined by its topology, not by detailed circuit parameters. Our work provides a theoretical foundation for circuit-based systems biology modeling. We anticipate RACIPE to be a powerful tool to predict and decode circuit design principles in an unbiased manner, and to quantitatively evaluate the robustness and heterogeneity of gene expression.

Author summary

Cells are able to robustly carry out their essential biological functions, possibly because of multiple layers of tight regulation via complex, yet well-designed, gene regulatory networks involving a substantial number of genes. State-of-the-art genomics technology has enabled the mapping of these large gene networks, yet it remains a tremendous challenge to elucidate their design principles and the regulatory mechanisms underlying their biological functions such as signal processing and decision-making. One of the key barriers is the absence of accurate kinetics for the regulatory interactions, especially from in vivo experiments. To this end, we have developed a new computational modeling method, Random Circuit Perturbation (RACIPE), to explore the dynamic behaviors of gene regulatory circuits without the requirement of detailed kinetic parameters. RACIPE takes a network topology as the input, and generates an unbiased ensemble of models with varying kinetic parameters. Each model is subjected to simulation, followed by statistical analysis for the ensemble. We tested RACIPE on several gene circuits, and found that the predicted gene expression patterns from all of the models converge to experimentally observed gene state clusters. We expect RACIPE to be a powerful method to identify the role of network topology in determining network operating principles.