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      PAGA: graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells

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          Abstract

          Single-cell RNA-seq quantifies biological heterogeneity across both discrete cell types and continuous cell transitions. Partition-based graph abstraction (PAGA) provides an interpretable graph-like map of the arising data manifold, based on estimating connectivity of manifold partitions ( https://github.com/theislab/paga). PAGA maps preserve the global topology of data, allow analyzing data at different resolutions, and result in much higher computational efficiency of the typical exploratory data analysis workflow. We demonstrate the method by inferring structure-rich cell maps with consistent topology across four hematopoietic datasets, adult planaria and the zebrafish embryo and benchmark computational performance on one million neurons.

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          The online version of this article (10.1186/s13059-019-1663-x) contains supplementary material, which is available to authorized users.

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          Most cited references11

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          Characterization of the single-cell transcriptional landscape by highly multiplex RNA-seq.

          Our understanding of the development and maintenance of tissues has been greatly aided by large-scale gene expression analysis. However, tissues are invariably complex, and expression analysis of a tissue confounds the true expression patterns of its constituent cell types. Here we describe a novel strategy to access such complex samples. Single-cell RNA-seq expression profiles were generated, and clustered to form a two-dimensional cell map onto which expression data were projected. The resulting cell map integrates three levels of organization: the whole population of cells, the functionally distinct subpopulations it contains, and the single cells themselves-all without need for known markers to classify cell types. The feasibility of the strategy was demonstrated by analyzing the transcriptomes of 85 single cells of two distinct types. We believe this strategy will enable the unbiased discovery and analysis of naturally occurring cell types during development, adult physiology, and disease.
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            Extracting a Cellular Hierarchy from High-dimensional Cytometry Data with SPADE

            Multiparametric single-cell analysis is critical for understanding cellular heterogeneity. Despite recent technological advances in single-cell measurements, methods for analyzing high-dimensional single-cell data are often subjective, labor intensive and require prior knowledge of the biological system under investigation. To objectively uncover cellular heterogeneity from single-cell measurements, we present a novel computational approach, Spanning-tree Progression Analysis of Density-normalized Events (SPADE). We applied SPADE to cytometry data of mouse and human bone marrow. In both cases, SPADE organized cells in a hierarchy of related phenotypes that partially recapitulated well-described patterns of hematopoiesis. In addition, SPADE produced a map of intracellular signal activation across the landscape of human hematopoietic development. SPADE revealed a functionally distinct cell population, natural killer (NK) cells, without using any NK-specific parameters. SPADE is a versatile method that facilitates the analysis of cellular heterogeneity, the identification of cell types, and comparison of functional markers in response to perturbations.
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              Single-cell mapping of gene expression landscapes and lineage in the zebrafish embryo

              High-throughput mapping of cellular differentiation hierarchies from single-cell data promises to empower systematic interrogations of vertebrate development and disease. Here, we applied single-cell RNA sequencing to >92,000 cells from zebrafish embryos during the first day of development. Using a graph-based approach, we mapped a cell state landscape that describes axis patterning, germ layer formation, and organogenesis. We tested how clonally related cells traverse this landscape by developing a transposon-based barcoding approach ("TracerSeq") for reconstructing single-cell lineage histories. Clonally related cells were often restricted by the state landscape, including a case in which two independent lineages converge on similar fates. Cell fates remained restricted to this landscape in chordin-deficient embryos. We provide web-based resources for further analysis of the single-cell data.
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                Author and article information

                Contributors
                alex.wolf@helmholtz-muenchen.de
                fabian.theis@helmholtz-muenchen.de
                Journal
                Genome Biol
                Genome Biol
                Genome Biology
                BioMed Central (London )
                1474-7596
                1474-760X
                19 March 2019
                19 March 2019
                2019
                : 20
                : 59
                Affiliations
                [1 ]ISNI 0000 0004 0483 2525, GRID grid.4567.0, Helmholtz Center Munich – German Research Center for Environmental Health, Institute of Computational Biology, ; Neuherberg, Munich, Germany
                [2 ]ISNI 0000000121885934, GRID grid.5335.0, Department of Haematology and Wellcome and Medical Research Council Cambridge Stem Cell Institute, , University of Cambridge, ; Cambridge, UK
                [3 ]ISNI 0000 0001 1014 0849, GRID grid.419491.0, Berlin Institute for Medical Systems Biology, Max-Delbrück Center for Molecular Medicine, ; Berlin, Germany
                [4 ]ISNI 0000 0000 9241 5705, GRID grid.24381.3c, Department of Medicine, Karolinska Institutet and Karolinska University Hospital, ; Stockholm, Sweden
                [5 ]ISNI 0000000123222966, GRID grid.6936.a, Department of Mathematics, Technische Universität München, ; Munich, Germany
                Author information
                http://orcid.org/0000-0002-8760-7838
                Article
                1663
                10.1186/s13059-019-1663-x
                6425583
                30890159
                78469211-bed0-42f0-b5a0-4ffcaff9ece9
                © The Author(s) 2019

                Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 5 November 2018
                : 26 February 2019
                Categories
                Method
                Custom metadata
                © The Author(s) 2019

                Genetics
                Genetics

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