Inference and analysis of cell-cell communication using CellChat

Understanding global communications among cells requires accurate representation of cell-cell signaling links and effective systems-level analyses of those links. We constructed a database of interactions among ligands, receptors and their cofactors that accurately represents known heteromeric molecular complexes. Based on mass action models, we then developed CellChat, a tool that is able to quantitively infer and analyze intercellular communication networks from single-cell RNA-sequencing (scRNA-seq) data. CellChat predicts major signaling inputs and outputs for cells and how those cells and signals coordinate for functions using network analysis and pattern recognition approaches. Through manifold learning and quantitative contrasts, CellChat classifies signaling pathways and delineates conserved and context-specific pathways across different datasets. Applications of CellChat to several mouse skin scRNA-seq datasets for embryonic development and adult wound healing shows its ability to extract complex signaling patterns, both previously known as well as novel. Our versatile and easy-to-use toolkit CellChat and a web-based Explorer (http://www.cellchat.org/) will help discover novel intercellular communications and build a cell-cell communication atlas in diverse tissues.

[1]  Leland McInnes,et al.  UMAP: Uniform Manifold Approximation and Projection , 2018, J. Open Source Softw..

[2]  Long Cai,et al.  Giotto, a pipeline for integrative analysis and visualization of single-cell spatial transcriptomic data , 2019, bioRxiv.

[3]  A. Danchin The specification of the immune response revisited , 1982, Survey of immunologic research.

[4]  Mauro J. Muraro,et al.  Dermal Condensate Niche Fate Specification Occurs Prior to Formation and Is Placode Progenitor Dependent. , 2019, Developmental cell.

[5]  David van Dijk,et al.  Manifold learning-based methods for analyzing single-cell RNA-sequencing data , 2018 .

[6]  C. Dickson,et al.  A crucial role for Fgfr2-IIIb signalling in epidermal development and hair follicle patterning , 2003, Development.

[7]  Ulrike von Luxburg,et al.  A tutorial on spectral clustering , 2007, Stat. Comput..

[8]  Qing Nie,et al.  Inferring spatial and signaling relationships between cells from single cell transcriptomic data , 2020, Nature Communications.

[9]  Jean-Michel Marin,et al.  Confidence bands for Brownian motion and applications to Monte Carlo simulation , 2007, Stat. Comput..

[10]  J. Bernhagen,et al.  MIF: a key player in cutaneous biology and wound healing , 2011, Experimental dermatology.

[11]  Bonnie Berger,et al.  Geometric Sketching Compactly Summarizes the Single-Cell Transcriptomic Landscape. , 2019, Cell systems.

[12]  D. D. Cros Fibroblast growth factor and epidermal growth factor in hair development. , 1993 .

[13]  Jean-Loup Guillaume,et al.  Fast unfolding of communities in large networks , 2008, 0803.0476.

[14]  Kan Liu,et al.  Giotto, a toolbox for integrative analysis and visualization of spatial expression data , 2020 .

[15]  T. Doetschman,et al.  The TGF-beta2 isoform is both a required and sufficient inducer of murine hair follicle morphogenesis. , 1999, Developmental biology.

[16]  Y. Kluger,et al.  Single-Cell Analysis Reveals a Hair Follicle Dermal Niche Molecular Differentiation Trajectory that Begins Prior to Morphogenesis. , 2019, Developmental cell.

[17]  T. Koh,et al.  Blocking Interleukin-1β Induces a Healing-Associated Wound Macrophage Phenotype and Improves Healing in Type 2 Diabetes , 2013, Diabetes.

[18]  Altuna Akalin,et al.  netSmooth: Network-smoothing based imputation for single cell RNA-seq , 2017, bioRxiv.

[19]  Amos Tanay,et al.  Dissecting cellular crosstalk by sequencing physically interacting cells , 2020, Nature Biotechnology.

[20]  Mirjana Efremova,et al.  CellPhoneDB: inferring cell–cell communication from combined expression of multi-subunit ligand–receptor complexes , 2020, Nature Protocols.

[21]  W. Border,et al.  Transforming Growth Factor β in Tissue Fibrosis , 1994 .

[22]  Panos M. Pardalos,et al.  Quantification of network structural dissimilarities , 2017, Nature Communications.

[23]  Shihua Zhang,et al.  A General Joint Matrix Factorization Framework for Data Integration and Its Systematic Algorithmic Exploration , 2020, IEEE Transactions on Fuzzy Systems.

[24]  B. Hinz,et al.  Wound‐healing defect of CD18−/− mice due to a decrease in TGF‐β1 and myofibroblast differentiation , 2005, The EMBO journal.

[25]  Darren J. Burgess,et al.  Spatial transcriptomics coming of age , 2019, Nature Reviews Genetics.

[26]  Benjamin J. Raphael,et al.  Network propagation: a universal amplifier of genetic associations , 2017, Nature Reviews Genetics.

[27]  N. Kaminski,et al.  WNT5A is a regulator of fibroblast proliferation and resistance to apoptosis. , 2009, American journal of respiratory cell and molecular biology.

[28]  G. Christensen,et al.  Wnt5a is elevated in heart failure and affects cardiac fibroblast function , 2017, Journal of Molecular Medicine.

[29]  R. Atit,et al.  Dermal β-catenin activity in response to epidermal Wnt ligands is required for fibroblast proliferation and hair follicle initiation , 2012, Development.

[30]  Michael J. T. Stubbington,et al.  Single-cell transcriptomics to explore the immune system in health and disease , 2017, Science.

[31]  A. Rezza,et al.  Wnt/β-catenin signaling in dermal condensates is required for hair follicle formation. , 2014, Developmental biology.

[32]  Benjamin D. Yu,et al.  Negative regulation of Shh levels by Kras and Fgfr2 during hair follicle development. , 2013, Developmental biology.

[33]  R. Galiano,et al.  Macrophage colony-stimulating factor accelerates wound healing and upregulates TGF-beta1 mRNA levels through tissue macrophages. , 1997, The Journal of surgical research.

[34]  J. Albina,et al.  Disruption of interleukin-1 signaling improves the quality of wound healing. , 2009, The American journal of pathology.

[35]  W. Hsu,et al.  Epidermal Wnt controls hair follicle induction by orchestrating dynamic signaling crosstalk between the epidermis and dermis , 2012, The Journal of investigative dermatology.

[36]  J. Weiss,et al.  Osteopontin and the skin: multiple emerging roles in cutaneous biology and pathology , 2009, Experimental dermatology.

[37]  Angela M. Christiano,et al.  KGF and EGF signalling block hair follicle induction and promote interfollicular epidermal fate in developing mouse skin , 2009, Development.

[38]  Kerstin B. Meyer,et al.  Single-cell reconstruction of the early maternal–fetal interface in humans , 2018, Nature.

[39]  M J Banda,et al.  Wound macrophages express TGF-alpha and other growth factors in vivo: analysis by mRNA phenotyping. , 1988, Science.

[40]  S. Millar,et al.  WNT signals are required for the initiation of hair follicle development. , 2002, Developmental cell.

[41]  Matthew J Ford,et al.  Hierarchical patterning modes orchestrate hair follicle morphogenesis , 2017, PLoS biology.

[42]  Douglas A. Lauffenburger,et al.  Analysis of Single-Cell RNA-Seq Identifies Cell-Cell Communication Associated with Tumor Characteristics , 2018, Cell reports.

[43]  Q. Nie,et al.  scAI: an unsupervised approach for the integrative analysis of parallel single-cell transcriptomic and epigenomic profiles , 2020, Genome Biology.

[44]  M. Mikkola,et al.  Fgf20 governs formation of primary and secondary dermal condensations in developing hair follicles. , 2013, Genes & development.

[45]  T. Shaw,et al.  Molecular mechanisms linking wound inflammation and fibrosis: knockdown of osteopontin leads to rapid repair and reduced scarring , 2008, The Journal of experimental medicine.

[46]  M. Mikkola,et al.  Hair follicle dermal condensation forms via Fgf20 primed cell cycle exit, cell motility, and aggregation , 2018, eLife.

[47]  Andrea Landherr,et al.  A Critical Review of Centrality Measures in Social Networks , 2010, Bus. Inf. Syst. Eng..

[48]  Jacques Colinge,et al.  SingleCellSignalR: inference of intercellular networks from single-cell transcriptomics , 2020, Nucleic acids research.

[49]  Qing Nie,et al.  Single-cell analysis reveals fibroblast heterogeneity and myeloid-derived adipocyte progenitors in murine skin wounds , 2019, Nature Communications.

[50]  Y. Saeys,et al.  NicheNet: modeling intercellular communication by linking ligands to target genes , 2019, Nature Methods.

[51]  R. Satija,et al.  Integrative single-cell analysis , 2019, Nature Reviews Genetics.

[52]  Randy L. Johnson,et al.  The signaling protein Wnt5a promotes TGFβ1-mediated macrophage polarization and kidney fibrosis by inducing the transcriptional regulators Yap/Taz , 2018, The Journal of Biological Chemistry.

[53]  Xianwen Ren,et al.  Reconstruction of cell spatial organization based on ligand-receptor mediated self-assembly , 2020, bioRxiv.

[54]  E. Birney,et al.  Mapping identifiers for the integration of genomic datasets with the R/Bioconductor package biomaRt , 2009, Nature Protocols.

[55]  M. Horan,et al.  Estrogen modulates cutaneous wound healing by downregulating macrophage migration inhibitory factor. , 2003, The Journal of clinical investigation.

[56]  P. Murphy,et al.  Chemokine Receptor CX3CR1 Mediates Skin Wound Healing by Promoting Macrophage and Fibroblast Accumulation and Function1 , 2008, The Journal of Immunology.

[57]  Elaine Fuchs,et al.  A Signaling Pathway Involving TGF-β2 and Snail in Hair Follicle Morphogenesis , 2004, PLoS biology.

[58]  D. Foreman,et al.  Neutralising antibody to TGF-beta 1,2 reduces cutaneous scarring in adult rodents. , 1994, Journal of cell science.

[59]  Carter T. Butts,et al.  Social Network Analysis with sna , 2008 .

[60]  W. Ge,et al.  Single-cell Transcriptome Profiling reveals Dermal and Epithelial cell fate decisions during Embryonic Hair Follicle Development , 2019, bioRxiv.

[61]  H. Bazzi,et al.  Transcriptional profiling of developing mouse epidermis reveals novel patterns of coordinated gene expression , 2007, Developmental dynamics : an official publication of the American Association of Anatomists.

[62]  Ka-Wai Mok,et al.  An updated classification of hair follicle morphogenesis , 2019, Experimental dermatology.

[63]  J. Ajani,et al.  iTALK: an R Package to Characterize and Illustrate Intercellular Communication , 2019, bioRxiv.

[64]  Damian Szklarczyk,et al.  STRING v11: protein–protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets , 2018, Nucleic Acids Res..

[65]  R. Derynck,et al.  Specificity, versatility, and control of TGF-β family signaling , 2019, Science Signaling.

[66]  B. Hogan,et al.  Altered wound healing in mice lacking a functional osteopontin gene (spp1). , 1998, The Journal of clinical investigation.

[67]  M. Elowitz,et al.  Challenges and emerging directions in single-cell analysis , 2017, Genome Biology.

[68]  Tao Peng,et al.  scEpath: energy landscape-based inference of transition probabilities and cellular trajectories from single-cell transcriptomic data , 2018, Bioinform..

[69]  Y. Kluger,et al.  Single-cell connectomic analysis of adult mammalian lungs , 2019, Science Advances.

[70]  E. Fuchs,et al.  Programming gene expression in developing epidermis. , 1994, Development.

[71]  F. Watt,et al.  Skin Cell Heterogeneity in Development, Wound Healing, and Cancer , 2018, Trends in cell biology.

[72]  Minoru Kanehisa,et al.  KEGG: new perspectives on genomes, pathways, diseases and drugs , 2016, Nucleic Acids Res..

[73]  Qing Nie,et al.  Cell lineage and communication network inference via optimization for single-cell transcriptomics , 2019, Nucleic acids research.

[74]  H. Sebastian Seung,et al.  Learning the parts of objects by non-negative matrix factorization , 1999, Nature.

[75]  Leland McInnes,et al.  UMAP: Uniform Manifold Approximation and Projection for Dimension Reduction , 2018, ArXiv.

[76]  C. Chuong,et al.  STAT3 signalling pathway is implicated in keloid pathogenesis by preliminary transcriptome and open chromatin analyses , 2019, Experimental dermatology.

[77]  W. Eaglstein,et al.  Interleukin-1 enhances epidermal wound healing. , 1990, Lymphokine research.