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MacArthur, B. D.

Publications and source records attributed to MacArthur, B. D..

14 recordsLinked to original sources

Charting cellular differentiation trajectories with Ricci flow

Complex biological processes, such as cellular differentiation, require an intricate rewiring of intra-cellular signalling networks. Previous characterisations of these networks revealed that promiscuity in signalling, quantified by a raised network entropy, underlies a less differentiated and malignant cell state. A theoretical connection between entropy and Ricci curvature has led to applications of discrete curvatures to characterise biological signalling networks at distinct time points during differentiation and malignancy. However, understanding and predicting the dynamics of biological network rewiring remains an open problem. Here we construct a framework to apply discrete Ricci curvature and Ricci flow to the problem of biological network rewiring. By investigating the relationship between network entropy and Forman-Ricci curvature, both theoretically and empirically on single-cell RNA-sequencing data, we demonstrate that the two measures do not always positively correlate, as has been previously suggested, and provide complementary rather than interchangeable information. We next employ discrete normalised Ricci flow, to derive network rewiring trajectories from transcriptomes of stem cells to differentiated cells, which accurately predict true intermediate time points of gene expression time courses. In summary, we present a differential geometry toolkit for investigation of dynamic network rewiring during cellular differentiation and cancer.

bioinformatics↗

Dual Dean entrainment with volume ratio modulation for efficient co-encapsulation: Extreme single-cell indexing

The future of single cell diversity screens involves ever-larger sample sizes, dictating the need for higher throughput methods with low analytical noise to accurately describe the nature of the cellular system. Current approaches are limited by the Poisson statistic, requiring dilute cell suspensions and associated losses in throughput. In this contribution, we apply Dean entrainment to both cell and bead inputs, defining different volume packets to effect efficient co-encapsulation. Volume ratio scaling was explored to identify optimal conditions. This enabled the co-encapsulation of single cells with reporter beads at rates of ~1 million cells/hour, while increasing assay signal-to-noise with cell multiplet rates of ~2.5% and capturing ~70% of cells. The method, called Pirouette-seq, extends our capacity to investigate biological systems. TOC AbstractPirouette-seq involves cell and reporter bead inertial ordering for efficient co-encapsulation, achieving a throughput of 1 million cells/hour, a 2.5% multiplet rate and a 70% cell capture efficiency.

cell biology↗

Measuring the Information Obtained from a Single-Cell Sequencing Experiment

Single-cell sequencing (sc-Seq) experiments are producing increasingly large data sets. However, large data sets do not necessarily contain large amounts of information. Here, we formally quantify the information obtained from a sc-Seq experiment and show that it corresponds to an intuitive notion of gene expression heterogeneity. We demonstrate a natural relation between our notion of heterogeneity and that of cell type, decomposing heterogeneity into that component attributable to differential expression between cell types (inter-cluster heterogeneity) and that remaining (intra-cluster heterogeneity). We test our definition of heterogeneity as the objective function of a clustering algorithm, and show that it is a useful descriptor for gene expression patterns associated with different cell types. Thus, our definition of gene heterogeneity leads to a biologically meaningful notion of cell type, as groups of cells that are statistically equivalent with respect to their patterns of gene expression. Our measure of heterogeneity, and its decomposition into inter- and intra-cluster, is non-parametric, intrinsic, unbiased, and requires no additional assumptions about expression patterns.

genomics↗

Heterogeneity and 'memory in stem cell populations

Modern single cell experiments have revealed unexpected heterogeneity in apparently functionally pure cell populations. However, we are still lacking a conceptual framework to understand this heterogeneity. Here, we propose that cellular memories - the ability of individual cells to record their developmental past and adapt their response to their environment accordingly - are an essential ingredient in any such theory. We illustrate this idea by considering a simple age-structured model of stem cell proliferation. Using this model we argue that heterogeneity is central to stem cell population function, and memories naturally explain why stem cell numbers increase through life, yet regenerative potency simultaneously declines.

cell biology↗

Single cell RNA sequence analysis of human bone marrow samples reveals new targets for isolation of skeletal stem cells using DNA-coated gold nanoparticles

There is a wealth of data indicating human bone marrow derived stromal cells (HBMSCs) contain the skeletal stem cell (SSC) with the potential to differentiate along the stromal osteogenic, adipogenic and chondrogenic lineages. However, despite these advances, current methods to isolate skeletal stem cells (SSCs) from human tissues have proved challenging as no single specific marker has been identified limiting understanding of SSC fate, immunophenotype and the widespread clinical application of these cells. While a number of cell surface markers can enrich for SSCs, none of the proposed markers, alone, provide a platform to isolate single cells with the ability to form bone, cartilage, and adipose tissue in humans. The current study details the application of oligonucleotide-coated nanoparticles, spherical nucleic acids (SNAs), to rapidly isolate human cells using mRNAs signatures detected in SSCs in real time, to identify stem and progenitor skeletal populations using single cell RNA sequencing. Based on scRNA-seq of samples from 11 patients, this method was able to identify novel targets for SSC enrichment, which were assessed in a total of 80 patients. This methodology was able to isolate potential SSCs found at a frequency of <1 in 1,000,000 in human bone marrow, with a capacity for tri-lineage differentiation in vitro. The current approach provides new targets and a platform to advance SSC isolation, enrichment with significant therapeutic impact therein.

cell biology↗

Universal principles of lineage architecture and stem cell identity in renewing tissues

Adult tissues in multicellular organisms typically contain a variety of stem, progenitor and differentiated cell types arranged in a lineage hierarchy that regulates healthy tissue turnover and repair. Lineage hierarchies in disparate tissues often exhibit common features, yet the general principles regulating their architecture are not known. Here, we provide a formal framework for understanding the relationship between cell molecular states (patterns of gene, protein expression etc. in the cell) and cell types that uses notions from network science to decompose the structure of cell state trajectories into functional units. Using this framework we show that many widely experimentally observed features of cell lineage architectures - including the fact that a single adult stem cell type always resides at the apex of a lineage hierarchy - arise as a natural consequence of homeostasis, and indeed are the only possible way that lineage architectures can be constructed to support homeostasis in renewing tissues. Furthermore, under suitable feedback regulation, for example from the stem cell niche, we show that the property of stemness is entirely determined by the cell environment. Thus, we argue that stem cell identities are contextual and not determined by hard-wired, cell-intrinsic, characteristics.

systems biology↗

Mapping biology from mouse to man using transfer learning

Biomedical research often involves conducting experiments on model organisms in the anticipation that the biology learnt from these experiments will transfer to the human. Yet, it is commonly the case that biology does not transfer effectively, often for unknown reasons. Despite its importance to translational research this transfer process is not currently rigorously quantified. Here, we show that transfer learning - the branch of machine learning that concerns passing information from one domain to another - can be used to efficiently map biology from mouse to man, using the bone marrow (BM) as a representative example of a complex tissue. We first trained an artificial neural network (ANN) to accurately recognize various different cell types in mouse BM using data obtained from single-cell RNA-sequencing (scRNA-Seq) experiments. We found that this ANN, trained exclusively on mouse data, was able to identify individual human cells obtained from comparable scRNA-Seq experiments of human BM with 83% overall accuracy. However, while some human cell types were easily identified, others were not, indicating important differences in biology. To obtain a more accurate map of the human BM we then retrained the mouse ANN using scRNA-Seq data from a limited sample of human BM cells. Typically, less than 10 human cells of a given type were needed to accurately learn its representation in the updated model. In some cases, human cell identities could be inferred directly from the mouse ANN without retraining, via a process of biologically-guided zero-shot learning. These results show how machine learning can be used to reconstruct complex biology from limited data and have broad implications for biomedical research.

systems biology↗

Genomic programming of antigen cross-presentation in IRF4-expressing human Langerhans cells

Langerhans cells (LCs) in the epidermis present MHC I and MHC II-restricted antigens thereby priming either CD8 or CD4 T cell immune responses. The genomic programs and transcription factors regulating antigen presentation in LCs remain to be elucidated. We show human LCs are highly efficient in MHC I-antigen cross-presentation but lack the transcription factor IRF8 that is critical in dendritic cells. LC migration from the epidermis enhances their ability to cross-present antigens and is accompanied by the induction of the transcription factor IRF4, whose expression is correlated by scRNA-seq with genes involved in ubiquitin-dependent protein degradation. Chromatin profiling reveals enrichment of EICE and AICE composite DNA binding motifs in regulatory regions of antigen-presentation genes, which can be recognized by IRF4 in conjunction with PU.1 or BATF3 expressed in LCs. Thus, the genomic programming of human LCs including inducible expression of IRF4 with enhanced cross-presentation distinguishes them from conventional dendritic cells.

immunology↗

A Topological Data Analysis Network Model of Asthma Based on Blood Gene Expression Profiles

Stratified medicine requires discretisation of disease populations for targeted treatments. We have developed and applied a discrete Morse theory clustering algorithm to a Topological Data Analysis (TDA) network model of 498 gene expression profiles of peripheral blood from asthma and healthy participants. The Morse clustering algorithm defined nine clusters, BC1-9, representing molecular phenotypes with discrete phenotypes including Type-1, 2 & 17 cytokine inflammatory pathways. The TDA network model and clusters were also characterised by activity of glucocorticoid receptor signalling associated with different expression profiles of glucocorticoid receptor (GR), according to microarray probesets targeted to the start or end of the GR mRNAs 3 UTR; suggesting differential GR mRNA processing as a possible driver of asthma phenotypes including steroid insensitivity.

molecular biology↗

Cell division history determines hematopoietic stem cell potency

Loss of stem cell self-renewal may underpin aging. Here, we combined single cell profiling, deep-learning, mathematical modelling and in vivo functional studies to explore how hematopoietic stem cell (HSC) division patterns evolve with age. We trained an artificial neural network (ANN) to accurately identify cell types in the hematopoietic hierarchy and predict their age from their gene expression patterns. We then used this ANN to compare daughter cell identities immediately after HSC divisions, and found that HSC self-renewal declines sharply with age. Furthermore, while HSC cell divisions are deterministic and intrinsically-regulated in young and old age, they become stochastic and niche-sensitive in mid-life. Analysis of evolving division patterns indicated that the self-renewal ability an individual HSC depends upon number times it has divided previously. We propose a model of HSC proliferation that accurately predicts the accumulation of HSCs with age, and reconciles the stochastic and instructive views of fate commitment.

cell biology↗

GenePy - a score for estimating gene pathogenicity in individuals using next-generation sequencing data

NGS is a revolutionising diagnosis and treatment of rare diseases. However, its relatively modest application in common diseases is limited by analytical approaches.\n\nInstead of variant-level approaches, typical for rare disease or large cohort analyses, contemporary investigation of common polygenic disorders requires the development of tools combining mutational burden and biological impact of a personalised set of mutations into single gene scores. GenePy (https://github.com/UoS-HGIG/GenePy) is a gene score for transforming sequencing data capable of estimating whole-gene pathogenicity on a per-patient basis.\n\nGenePy implements known deleteriousness metrics, incorporates allele frequency and individual zygosity information. Individuals harbouring multiple rare highly deleterious mutations accumulate extreme gene scores while the majority of genes usually achieve very low scores. Following correction for gene length, GenePy intuitively prioritises genes within individuals and affords gene/pathway score comparison between groups of individuals. Herein, we generate GenePy scores from whole-exome sequencing data for [~]15,000 genes across a cohort of 508 individuals. We describe score attributes and model behaviour under various biological conditions.\n\nWe demonstrate proof of concept that GenePy sensitively identifies known causal genes by calculating GenePy scores for NOD2 (an established causal Crohns Disease gene), in a modest cohort of patients for comparison against controls. This test of GenePy using a positive control gene demonstrates markedly more significant results (p=1.37 x 10-4) compared to the most commonly applied tool for combining common and rare variation.\n\nIn addition to increasing the biological information content for each variant, the per gene-per individual nature of GenePy transforms the utility of sequencing data. GenePy scores are intuitive when assessing for individual patients or for comparing between groups. Because GenePy intrinsically reflects pathogenicity at the gene level, this specifically facilitates downstream data integration (e.g. into machine learning, network and topological analyses) with transcriptomic and proteomic data that also report at the gene level.\n\nAuthor SummaryRapid technological advances have made DNA sequencing an effective, economic tool for detecting genomic variation. Detecting rare variation at the individual level is proving very successful in identifying the genetic causes of disease when just single mutations are sufficient to manifest disease. However, interpreting genomic data is much less straightforward for common diseases such as asthma, arthritis or heart disease where many genetic changes across multiple genes combine with the environment to bring about disease symptoms.\n\nWe have developed a new scoring system called GenePy that generates whole gene pathogenicity scores for indiviual patients. The score corrects for the length of the gene and is intuitive to use. Unlike many mutation deleteriousness metrics, GenePy also takes into account the population frequency of the variant and the number of copies of any given mutation and combines data for as many variants as are present in a given gene for any one individual.\n\nIn this paper we apply the GenePy scoring system to a cohort of over 500 individuals for whom we have sequencing data across all genomics regions that code for protein. We descibe how GenePy performs and demonstrate superior sensitivity to detect known causal genes in a common autoimmune condition.

genomics↗

Pattern analysis of pluripotency reveals the molecular basis of nai&#x0308;ve, primed and formative states

The molecular regulatory network underlying stem cell pluripotency has been intensively studied, and we now have a reliable ensemble model for the average pluripotent cell. However, evidence of significant cell-to-cell variability suggests that the activity of this network varies within individual stem cells, leading to differential processing of environmental signals and variability in cell fates. Here, we adapt a method originally designed for face recognition to infer regulatory network patterns within individual cells from single-cell expression data. Using this method we identify three distinct network configurations in cultured mouse embryonic stem cells - corresponding to naive and formative pluripotent states and an early primitive endoderm state - and associate these configurations with particular combinations of regulatory network activity archetypes that govern different aspects of the cells response to environmental stimuli, cell cycle status and core information processing circuitry. These results show how variability in cell identities arise naturally from alterations in underlying regulatory network dynamics and demonstrate how methods from machine learning may be used to better understand single cell biology, and the collective dynamics of cell communities.

systems biology↗

Information Theory and Stem Cell Biology

Purpose of ReviewTo outline how ideas from Information Theory may be used to analyze single cell data and better understand stem cell behaviour.\n\nRecent findingsRecent technological breakthroughs in single cell profiling have made it possible to interrogate cell-to-cell variability in a multitude of contexts, including the role it plays in stem cell dynamics. Here we review how measures from information theory are being used to extract biological meaning from the complex, high-dimensional and noisy datasets that arise from single cell profiling experiments. We also discuss how concepts linking information theory and statistical mechanics are being used to provide insight into cellular identity, variability and dynamics.\n\nSummaryWe provide a brief introduction to some basic notions from information theory and how they may be used to understand stem cell identities at the single cell level. We also discuss how work in this area might develop in the near future.

cell biology↗

Stem cell differentiation is a stochastic process with memory

Pluripotent stem cells are able to self-renew indefinitely in culture and differentiate into all somatic cell types in vivo. While much is known about the molecular basis of pluripotency, the molecular mechanisms of lineage commitment are complex and only partially understood. Here, using a combination of single cell profiling and mathematical modeling, we examine the differentiation dynamics of individual mouse embryonic stem cells (ESCs) as they progress from the ground state of pluripotency along the neuronal lineage. In accordance with previous reports we find that cells do not transit directly from the pluripotent state to the neuronal state, but rather first stochastically permeate an intermediate primed pluripotent state, similar to that found in the maturing epiblast in development. However, analysis of rate at which individual cells enter and exit this intermediate metastable state using a hidden Markov model reveals that the observed ESC and epiblast-like macrostates conceal a chain of unobserved cellular microstates, which individual cells transit through stochastically in sequence. These hidden microstates ensure that individual cells spend well-defined periods of time in each functional macrostate and encode a simple form of epigenetic memory that allows individual cells to record their position on the differentiation trajectory. To examine the generality of this model we also consider the differentiation of mouse hematopoietic stem cells along the myeloid lineage and observe remarkably similar dynamics, suggesting a general underlying process. Based upon these results we suggest a statistical mechanics view of cellular identities that distinguishes between functionally-distinct macrostates and the many functionally-similar molecular microstates associated with each macrostate. Taken together these results indicate that differentiation is a discrete stochastic process amenable to analysis using the tools of statistical mechanics.

systems biology↗