Ilya Shmulevich

From Boolean to probabilistic Boolean networks as models of genetic regulatory networks

Mathematical and computational modeling of genetic regulatory networks promises to uncover the fundamental principles governing biological systems in an integrative and holistic manner. It also paves the way toward the development of systematic approaches for effective therapeutic intervention in disease. The central theme in this paper is the Boolean formalism as a building block for modeling complex, large-scale, and dynamical networks of genetic interactions. We discuss the goals of modeling genetic networks as well as the data requirements. The Boolean formalism is justified from several points of view. We then introduce Boolean networks and discuss their relationships to nonlinear digital filters. The role of Boolean networks in understanding cell differentiation and cellular functional states is discussed. The inference of Boolean networks from real gene expression data is considered from the viewpoints of computational learning theory and nonlinear signal processing, touching on computational complexity of learning and robustness. Then, a discussion of the need to handle uncertainty in a probabilistic framework is presented, leading to an introduction of probabilistic Boolean networks and their relationships to Markov chains. Methods for quantifying the influence of genes on other genes are presented. The general question of the potential effect of individual genes on the global dynamical network behavior is considered using stochastic perturbation analysis. This discussion then leads into the problem of target identification for therapeutic intervention via the development of several computational tools based on first-passage times in Markov chains. Examples from biology are presented throughout the paper.

On Learning Gene Regulatory Networks Under the Boolean Network Model

Boolean networks are a popular model class for capturing the interactions of genes and global dynamical behavior of genetic regulatory networks. Recently, a significant amount of attention has been focused on the inference or identification of the model structure from gene expression data. We consider the Consistency as well as Best-Fit Extension problems in the context of inferring the networks from data. The latter approach is especially useful in situations when gene expression measurements are noisy and may lead to inconsistent observations. We propose simple efficient algorithms that can be used to answer the Consistency Problem and find one or all consistent Boolean networks relative to the given examples. The same method is extended to learning gene regulatory networks under the Best-Fit Extension paradigm. We also introduce a simple and fast way of finding all Boolean networks having limited error size in the Best-Fit Extension Problem setting. We apply the inference methods to a real gene expression data set and present the results for a selected set of genes.

Binary analysis and optimization-based normalization of gene expression data

MOTIVATIONnMost approaches to gene expression analysis use real-valued expression data, produced by high-throughput screening technologies, such as microarrays. Often, some measure of similarity must be computed in order to extract meaningful information from the observed data. The choice of this similarity measure frequently has a profound effect on the results of the analysis, yet no standards exist to guide the researcher.nnnRESULTSnTo address this issue, we propose to analyse gene expression data entirely in the binary domain. The natural measure of similarity becomes the Hamming distance and reflects the notion of similarity used by biologists. We also develop a novel data-dependent optimization-based method, based on Genetic Algorithms (GAs), for normalizing gene expression data. This is a necessary step before quantizing gene expression data into the binary domain and generally, for comparing data between different arrays. We then present an algorithm for binarizing gene expression data and illustrate the use of the above methods on two different sets of data. Using Multidimensional Scaling, we show that a reasonable degree of separation between different tumor types in each data set can be achieved by working solely in the binary domain. The binary approach offers several advantages, such as noise resilience and computational efficiency, making it a viable approach to extracting meaningful biological information from gene expression data.

Differential gene and protein expression in primary breast malignancies and their lymph node metastases as revealed by combined cDNA microarray and tissue microarray analysis.

Metastatic disease is a major adverse prognostic factor in breast carcinoma. Lymph node metastases often represent the first step in the metastatic process.

Steady‐state analysis of genetic regulatory networks modelled by probabilistic Boolean networks

Probabilistic Boolean networks (PBNs) have recently been introduced as a promising class of models of genetic regulatory networks. The dynamic behaviour of PBNs can be analysed in the context of Markov chains. A key goal is the determination of the steady-state (long-run) behaviour of a PBN by analysing the corresponding Markov chain. This allows one to compute the long-term influence of a gene on another gene or determine the long-term joint probabilistic behaviour of a few selected genes. Because matrix-based methods quickly become prohibitive for large sizes of networks, we propose the use of Monte Carlo methods. However, the rate of convergence to the stationary distribution becomes a central issue. We discuss several approaches for determining the number of iterations necessary to achieve convergence of the Markov chain corresponding to a PBN. Using a recently introduced method based on the theory of two-state Markov chains, we illustrate the approach on a sub-network designed from human glioma gene expression data and determine the joint steadystate probabilities for several groups of genes.

CONTROL OF STATIONARY BEHAVIOR IN PROBABILISTIC BOOLEAN NETWORKS BY MEANS OF STRUCTURAL INTERVENTION

Probabilistic Boolean Networks (PBNs) were recently introduced as models of gene regulatory networks. The dynamical behavior of PBNs, which are probabilistic generalizations of Boolean networks, ca...

The role of certain Post classes in Boolean network models of genetic networks

A topic of great interest and debate concerns the source of order and remarkable robustness observed in genetic regulatory networks. The study of the generic properties of Boolean networks has proven to be useful for gaining insight into such phenomena. The main focus, as regards ordered behavior in networks, has been on canalizing functions, internal homogeneity or bias, and network connectivity. Here we examine the role that certain classes of Boolean functions that are closed under composition play in the emergence of order in Boolean networks. The closure property implies that any gene at any number of steps in the future is guaranteed to be governed by a function from the same class. By means of Derrida curves on random Boolean networks and percolation simulations on square lattices, we demonstrate that networks constructed from functions belonging to these classes have a tendency toward ordered behavior. Thus they are not overly sensitive to initial conditions, and damage does not readily spread throughout the network. In addition, the considered classes are significantly larger than the class of canalizing functions as the connectivity increases. The functions in these classes exhibit the same kind of preference toward biased functions as do canalizing functions, meaning that functions from this class are likely to be biased. Finally, functions from this class have a natural way of ensuring robustness against noise and perturbations, thus representing plausible evolutionarily selected candidates for regulatory rules in genetic networks.

Growing genetic regulatory networks from seed genes

MOTIVATIONnA number of models have been proposed for genetic regulatory networks. In principle, a network may contain any number of genes, so long as data are available to make inferences about their relationships. Nevertheless, there are two important reasons why the size of a constructed network should be limited. Computationally and mathematically, it is more feasible to model and simulate a network with a small number of genes. In addition, it is more likely that a small set of genes maintains a specific core regulatory mechanism.nnnRESULTSnSubnetworks are constructed in the context of a directed graph by beginning with a seed consisting of one or more genes believed to participate in a viable subnetwork. Functionalities and regulatory relationships among seed genes may be partially known or they may simply be of interest. Given the seed, we iteratively adjoin new genes in a manner that enhances subnetwork autonomy. The algorithm is applied using both the coefficient of determination and the Boolean-function influence among genes, and it is illustrated using a glioma gene-expression dataset.nnnAVAILABILITYnSoftware for the seed-growing algorithm will be available at the website for Probabilistic Boolean Networks: http://www2.mdanderson.org/app/ilya/PBN/PBN.htm

Steady-state probabilities for attractors in probabilistic boolean networks

Boolean networks form a class of disordered dynamical systems that have been studied in physics owing to their relationships with disordered systems in statistical mechanics and in biology as models of genetic regulatory networks. Recently they have been generalized to probabilistic Boolean networks (PBNs) to facilitate the incorporation of uncertainty in the model and to represent cellular context changes in biological modeling. In essence, a PBN is composed of a family of Boolean networks between which the PBN switches in a stochastic fashion. In whatever framework Boolean networks are studied, their most important attribute is their attractors. Left to run, a Boolean network will settle into one of a collection of state cycles called attractors. The set of states from which the network will transition into a specific attractor forms the basin of the attractor. The attractors represent the essential long-run behavior of the network. In a classical Boolean network, the network remains in an attractor once there; in a Boolean network with perturbation, the states form an ergodic Markov chain and the network can escape an attractor, but it will return to it or a different attractor unless interrupted by another perturbation; in a probabilistic Boolean network, so long as the PBN remains in one of its constituent Boolean networks it will behave as a Boolean network with perturbation, but upon a switch it will move to an attractor of the new constituent Boolean network. Given the ergodic nature of the model, the steady-state probabilities of the attractors are critical to network understanding. Heretofore they have been found by simulation; in this paper we derive analytic expressions for these probabilities, first for Boolean networks with perturbation and then for PBNs.

Insulin-like growth factor binding protein 2 promotes ovarian cancer cell invasion

BackgroundInsulin-like growth factor binding protein 2 (IGFBP2) is overexpressed in ovarian malignant tissues and in the serum and cystic fluid of ovarian cancer patients, suggesting an important role of IGFBP2 in the biology of ovarian cancer. The purpose of this study was to assess the role of increased IGFBP2 in ovarian cancer cells.ResultsUsing western blotting and tissue microarray analyses, we showed that IGFBP2 was frequently overexpressed in ovarian carcinomas compared with normal ovarian tissues. Furthermore, IGFBP2 was significantly overexpressed in invasive serous ovarian carcinomas compared with borderline serous ovarian tumors. To test whether increased IGFBP2 contributes to the highly invasive nature of ovarian cancer cells, we generated IGFBP2-overexpressing cells from an SKOV3 ovarian cancer cell line, which has a very low level of endogenous IGFBP2. A Matrigel invasion assay showed that these IGFBP2-overexpressing cells were more invasive than the control cells. We then designed small interference RNA (siRNA) molecules that attenuated IGFBP2 expression in PA-1 ovarian cancer cells, which have a high level of endogenous IGFBP2. The Matrigel invasion assay showed that the attenuation of IGFBP2 expression indeed decreased the invasiveness of PA-1 cells.ConclusionsWe therefore showed that IGFBP2 enhances the invasion capacity of ovarian cancer cells. Blockage of IGFBP2 may thus constitute a viable strategy for targeted cancer therapy.