Understanding the multicellular organization of stem cells is vital for determining the mechanisms that coordinate cell fate decision-making during differentiation; these mechanisms range from neighbor-to-neighbor communication to tissue-level biochemical gradients. Current methods for quantifying multicellular patterning tend to capture the spatial properties of cell colonies at a fixed scale and typically rely on human annotation. We present a computational pipeline that utilizes topological data analysis to generate quantitative, multiscale descriptors which capture the shape of data extracted from 2D multichannel microscopy images. By applying our pipeline to certain stem cell colonies, we detected subtle differences in patterning that reflect distinct spatial organization associated with loss of pluripotency. These results yield insight into putative directed cellular organization and morphogen-mediated, neighbor-to-neighbor signaling. Because of its broad applicability to immunofluorescence microscopy images, our pipeline is well-positioned to serve as a general-purpose tool for the quantitative study of multicellular pattern formation.

Boolean networks have been used in a variety of settings, as models for general complex systems as well as models of specific systems in diverse fields, such as biology, engineering, and computer science. Traditionally, their properties as dynamical systems have been studied through simulation studies, due to a lack of mathematical structure. This paper uses a common mathematical technique to identify a class of Boolean networks with a "simple" structure and describes an algorithm to construct arbitrary extensions of a collection of simple Boolean networks. In this way, all Boolean networks can be obtained from a collection of simple Boolean networks as building blocks. The paper furthermore provides a formula for the number of extensions of given simple networks and, in some cases, provides a parametrization of those extensions. This has potential applications to the construction of networks with particular properties, for instance in synthetic biology, and can also be applied to develop efficient control algorithms for Boolean network models.

Understanding how stem cells organize to form early tissue layers remains an important open question in developmental biology. Helpful in understanding this process are biomarkers or features that signal when a significant transition or decision occurs. We show such features from the spatial layout of the cells in a colony are sufficient to train neural networks to classify stem cell colonies according to differentiation protocol treatments each colony has received. We use topological data analysis to derive input information about the cells' positions to a four-layer feedforward neural network. We find that despite the simplicity of this approach, such a network has performance similar to the traditional image classifier ResNet. We also find that network performance may reveal the time window during which differentiation occurs across multiple conditions.

Over the past several decades, algebraic geometry has provided innovative approaches to biological experimental design that resolved theoretical questions and improved computational efficiency. However, guaranteeing uniqueness and perfect recovery of models are still open problems. In this work we study the problem of uniqueness of wiring diagrams. We use as a modeling framework polynomial dynamical systems and utilize the correspondence between simplicial complexes and square-free monomial ideals from Stanley-Reisner theory to develop theory and construct an algorithm for identifying input data sets $V\subset \mathbb F_p^n$ that are guaranteed to correspond to a unique minimal wiring diagram regardless of the experimental output. We apply the results on a tumor-suppression network mediated by epidermal derived growth factor receptor and demonstrate how careful experimental design decisions can lead to a unique minimal wiring diagram identification. One of the insights of the theoretical work is the connection between the uniqueness of a wiring diagram for a given $V\subset \mathbb F_p^n$ and the uniqueness of the reduced Gr\"obner basis of the polynomial ideal $I(V)\subset \mathbb F_p[x_1,\ldots, x_n]$. We discuss existing results and introduce a new necessary condition on the points in $V$ for uniqueness of the reduced Gr\"obner basis of $I(V)$. These results also point to the importance of the relative proximity of the experimental input points on the number of minimal wiring diagrams, which we then study computationally. We find that there is a concrete heuristic way to generate data that tends to result in fewer minimal wiring diagrams.

Due to cost concerns, it is optimal to gain insight into the connectivity of biological and other networks using as few experiments as possible. Data selection for unique network connectivity identification has been an open problem since the introduction of algebraic methods for reverse engineering for almost two decades. In this manuscript we determine what data sets uniquely identify the unsigned wiring diagram corresponding to a system that is discrete in time and space. Furthermore, we answer the question of uniqueness for signed wiring diagrams for Boolean networks. Computationally, unsigned and signed wiring diagrams have been studied separately, and in this manuscript we also show that there exists an ideal capable of encoding both unsigned and signed information. This provides a unified approach to studying reverse engineering that also gives significant computational benefits.

Understanding the temporal evolution of cells poses a significant challenge in developmental biology. This study embarks on a comparative analysis of various machine-learning techniques to classify sequences of cell colony images, thereby aiming to capture dynamic transitions of cellular states. Utilizing transfer learning with advanced classification networks, we achieved high accuracy in single-timestamp image categorization. We introduce temporal models—LSTM, R-Transformer, and ViViT—to explore the effectiveness of integrating temporal features in classification, comparing their performance against non-temporal models. This research benchmarks various machine learning approaches in understanding cellular dynamics, setting a foundation for future studies to enhance our understanding of cellular developments with computational methods, contributing significantly to biological research advancements.### Competing Interest StatementThe authors have declared no competing interest.

MOTIVATION:While there are software packages that analyze Boolean, ternary, or other multi-state models, none compute the complete state space of function-based models over any finite set. Results: We propose Cyclone, a simple light-weight software package which simulates the complete state space for a finite dynamical system over any finite set.AVAILABILITY AND IMPLEMENTATION:Source code is freely available at https://github.com/discretedynamics/cyclone under the Apache-2.0 license.

Boolean functions can be represented in many ways including logical forms, truth tables, and polynomials. Additionally, Boolean functions have different canonical representations such as minimal disjunctive normal forms. Another canonical representation is based on the polynomial representation of Boolean functions and the biologically motivated concept of canalization: any Boolean function can be written as a nested product of canalizing layers and a polynomial that contains the variables that are never canalizing. In this paper we study the problem of identifying the canalizing layers of a Boolean function. First, we show that the problem of finding the canalizing layers is NP-hard. Second, we present several algorithms for finding the canalizing layers, discuss their complexities, and compare their performances. Third, we exhibit how the canalizing layers format can be used to find a disjunctive normal form for any nested canalizing function. Another application deals with the reverse engineering of Boolean networks with a prescribed layering format. Implementations of the developed algorithms in Python and in the computer algebra system Macaulay2 are available at https://github.com/ckadelka/BooleanCanalization.

An agent-based model (ABM) is a computational model in which the local interactions of autonomous agents with each other and with their environment give rise to global properties within a given domain. As the detail and complexity of these models has grown, so too has the computational expense of running several simulations to perform sensitivity analysis and evaluate long-term model behavior. Here, we generalize a framework for mathematically formalizing ABMs to explicitly incorporate features commonly found in biological systems: appearance of agents (birth), removal of agents (death), and locally dependent state changes. We then use our broader framework to extend an approach for estimating long-term behavior without simulations, specifically changes in population densities over time. The approach is probabilistic and relies on treating the discrete, incremental update of an ABM via "time steps" as a Markov process to generate expected values for agents at each time step. As case studies, we apply our extensions to both a simple ABM based on the Game of Life and a published ABM of rib development in vertebrates.

Design of experiments and model selection are essential steps in biological data-science pipelines and coupling them has the potential to improve computational efficiency. In particular approaches which can handle data from unspecified distributions and nonlinear models are of interest. We propose an algebra-based computational framework to unify experimental design and model selection for discrete data sets and minimal polynomial models. We use an affine transformation, called a linear shift, to identify data sets and bases of polynomial models simultaneously. We present the theoretical foundation for a web-accessible database. This framework enables us to address two illustrative questions that may arise when studying biological systems: finding the data which identifies a set of known interactions and finding identifiable interactions given a set of data. As an example, we apply the methodology to a previously constructed pharmacodynamic model of epidermal derived growth factor receptor (EGFR) signaling.

Developing efficient computational methods to assess the impact of external interventions on the dynamics of a network model is an important problem in systems biology. This paper focuses on quantifying the global changes that result from the application of an intervention to produce a desired effect, which we define as the total effect of the intervention. The type of mathematical models that we will consider are discrete dynamical systems which include the widely used Boolean networks and their generalizations. The potential interventions can be represented by a set of nodes and edges that can be manipulated to produce a desired effect on the system. We use a class of regulatory rules called nested canalizing functions that frequently appear in published models and were inspired by the concept of canalization in evolutionary biology. In this paper, we provide a polynomial normal form based on the canalizing properties of regulatory functions. Using this polynomial normal form, we give a set of formulas for counting the maximum number of transitions that will change in the state space upon an edge deletion in the wiring diagram. These formulas rely on the canalizing structure of the target function since the number of changed transitions depends on the canalizing layer that includes the input to be deleted. We also present computations on random networks to compare the exact number of changes with the upper bounds provided by our formulas. Finally, we provide statistics on the sharpness of these upper bounds in random networks.

Many biological ecosystems exhibit chaotic behavior, demonstrated either analytically using parameter choices in an associated dynamical systems model or empirically through analysis of experimental data. In this paper, we use existing software tools (COPASI, R) to explore dynamical systems and uncover regions with positive Lyapunov exponents where thus chaos exists. We evaluate the ability of the software’s optimization algorithms to find these positive values with several dynamical systems used to model biological populations. The algorithms have been able to identify parameter sets which lead to positive Lyapunov exponents, even when those exponents lie in regions with small support. For one of the examined systems, we observed that positive Lyapunov exponents were not uncovered when executing a search over the parameter space with small spacings between values of the independent variables.

We present results and conjectures on the connection between the convexity of a neural code and the canonical form of its ideal. The connection is established through properties of the Gröbner basis of the neural ideal and the uniqueness of its reduced form. An efficient algorithm for identifying neural codes with unique reduced Gröbner bases is introduced.

In the context of modeling biological systems, it is of interest to generate ideals of points with a unique reduced Groebner basis, and the first main goal of this paper is to identify classes of ideals in polynomial rings which share this property. Moreover, we provide methodologies for constructing such ideals. We then relax the condition of uniqueness. The second and most relevant topic discussed here is to consider and identify pairs of ideals with the same number of reduced Groebner bases, that is, with the same cardinality of their associated Groebner fan.

Work by Costantino et al. (Am Assoc Adv Sci 275(5298):389–391, 1997) and Kot et al. (Bull Math Biol 54(4):619–648, 1992) demonstrate that chaotic behavior does occur in biological systems. We show that chaotic behavior can also be used to ensure the survival of the species involved in a system. We adopt the concept of permanence as a measure of survival and take advantage of present chaotic behavior to push a non-permanent system into permanence through a control algorithm. We apply the algorithm to a Lotka-Volterra type two-prey, one-predator model and a food chain model and demonstrate its effectiveness in taking advantage of chaotic behavior to achieve a desirable state for all species involved. In particular, we show that harvesting of the predator is a practical and effective control for insuring the thriving of all species in the system.

Model selection based on experimental data is an important challenge in biological data science. Particularly when collecting data is expensive or time-consuming, as it is often the case with clinical trial and biomolecular experiments, the problem of selecting information-rich data becomes crucial for creating relevant models. We identify geometric properties of input data that result in an unique algebraic model, and we show that if the data form a staircase, or a so-called linear shift of a staircase, the ideal of the points has a unique reduced Gröbner basis and thus corresponds to a unique model. We use linear shifts to partition data into equivalence classes with the same basis. We demonstrate the utility of the results by applying them to a Boolean model of the well-studied lac operon in E. coli.

BACKGROUND:At the molecular level, nonlinear networks of heterogeneous molecules control many biological processes, so that systems biology provides a valuable approach in this field, building on the integration of experimental biology with mathematical modeling. One of the biggest challenges to making this integration a reality is that many life scientists do not possess the mathematical expertise needed to build and manipulate mathematical models well enough to use them as tools for hypothesis generation. Available modeling software packages often assume some modeling expertise. There is a need for software tools that are easy to use and intuitive for experimentalists.RESULTS:This paper introduces PlantSimLab, a web-based application developed to allow plant biologists to construct dynamic mathematical models of molecular networks, interrogate them in a manner similar to what is done in the laboratory, and use them as a tool for biological hypothesis generation. It is designed to be used by experimentalists, without direct assistance from mathematical modelers.CONCLUSIONS:Mathematical modeling techniques are a useful tool for analyzing complex biological systems, and there is a need for accessible, efficient analysis tools within the biological community. PlantSimLab enables users to build, validate, and use intuitive qualitative dynamic computer models, with a graphical user interface that does not require mathematical modeling expertise. It makes analysis of complex models accessible to a larger community, as it is platform-independent and does not require extensive mathematical expertise.

Developing efficient computational methods to change the state of a cell from an undesirable condition, e.g. diseased, into a desirable, e.g. healthy, condition is an important goal of systems biology. The identification of potential interventions can be achieved through mathematical modeling of the state of a cell by finding appropriate input manipulations in the model that represent external interventions. This paper focuses on quantifying the unwanted or unplanned changes that come along with the application of an intervention to produce a desired effect, which we define as the \emph{side effects} of the intervention. The type of mathematical models that we will consider are discrete dynamical systems which include the widely used Boolean networks and their generalizations. The potential control targets can be represented by a set of nodes and edges that can be manipulated to produce a desired effect on the system. This paper presents practical tools along with applications for the analysis and control of multistate networks. The first result is a polynomial normal form representation for discrete functions that provides a partition of the inputs of the function into canalizing and non-canalizing variables and, within the canalizing ones, we categorize the input variables into layers of canalization. The second theoretical result is a set of formulas for counting the maximum number of transitions that will change in the state space upon an edge deletion in the wiring diagram. These formulas rely on the stratification of the inputs of the target function where the number of changed transitions depends on the layer of canalization that includes the input to be deleted. Applications from using these formulas to estimate the number of changes in the state space and comparisons with the actual number of changes are also presented.