Highly Dynamic Transcriptional Signature of Distinct Macrophage Subsets during Sterile Inflammation, Resolution, and Tissue Repair

Macrophage gene expression determines phagocyte responses and effector functions. Macrophage plasticity has been mainly addressed in in vitro models that do not account for the environmental complexity observed in vivo. In this study, we show that microarray gene expression profiling revealed a highly dynamic landscape of transcriptomic changes of Ly6CposCX3CR1lo and Ly6CnegCX3CR1hi macrophage populations during skeletal muscle regeneration after a sterile damage. Systematic gene expression analysis revealed that the time elapsed, much more than Ly6C status, was correlated with the largest differential gene expression, indicating that the time course of inflammation was the predominant driving force of macrophage gene expression. Moreover, Ly6Cpos/Ly6Cneg subsets could not have been aligned to canonical M1/M2 profiles. Instead, a combination of analyses suggested the existence of four main features of muscle-derived macrophages specifying important steps of regeneration: 1) infiltrating Ly6Cpos macrophages expressed acute-phase proteins and exhibited an inflammatory profile independent of IFN-γ, making them damage-associated macrophages; 2) metabolic changes of macrophages, characterized by a decreased glycolysis and an increased tricarboxylic acid cycle/oxidative pathway, preceded the switch to and sustained their anti-inflammatory profile; 3) Ly6Cneg macrophages, originating from skewed Ly6Cpos cells, actively proliferated; and 4) later on, restorative Ly6Cneg macrophages were characterized by a novel profile, indicative of secretion of molecules involved in intercellular communications, notably matrix-related molecules. These results show the highly dynamic nature of the macrophage response at the molecular level after an acute tissue injury and subsequent repair, and associate a specific signature of macrophages to predictive specialized functions of macrophages at each step of tissue injury/repair.

[1]  Michael D. Schneider,et al.  Monocyte/Macrophage-derived IGF-1 Orchestrates Murine Skeletal Muscle Regeneration and Modulates Autocrine Polarization. , 2015, Molecular therapy : the journal of the American Society of Gene Therapy.

[2]  F. Rossi,et al.  Nilotinib reduces muscle fibrosis in chronic muscle injury by promoting TNF-mediated apoptosis of fibro/adipogenic progenitors , 2015, Nature Medicine.

[3]  E. Abraham,et al.  Pyruvate Dehydrogenase Kinase 1 Participates in Macrophage Polarization via Regulating Glucose Metabolism , 2015, The Journal of Immunology.

[4]  R. Ransohoff,et al.  A dynamic spectrum of monocytes arising from the in situ reprogramming of CCR2+ monocytes at a site of sterile injury , 2015, The Journal of experimental medicine.

[5]  Abhishek K. Jha,et al.  Network integration of parallel metabolic and transcriptional data reveals metabolic modules that regulate macrophage polarization. , 2015, Immunity.

[6]  Matthew E. Ritchie,et al.  limma powers differential expression analyses for RNA-sequencing and microarray studies , 2015, Nucleic acids research.

[7]  J. Stender,et al.  Environment Drives Selection and Function of Enhancers Controlling Tissue-Specific Macrophage Identities , 2015, Cell.

[8]  I. Amit,et al.  Tissue-Resident Macrophage Enhancer Landscapes Are Shaped by the Local Microenvironment , 2014, Cell.

[9]  J. McQualter,et al.  SAA drives proinflammatory heterotypic macrophage differentiation in the lung via CSF‐1R‐dependent signaling , 2014, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[10]  S. Goerdt,et al.  Macrophage activation and polarization: nomenclature and experimental guidelines. , 2014, Immunity.

[11]  P. Libby,et al.  Chigh Monocytes Depend on Nr 4 a 1 to Balance Both Inflammatory and Reparative Phases in the Infarcted Myocardium , 2014 .

[12]  L. McManus,et al.  Altered macrophage phenotype transition impairs skeletal muscle regeneration. , 2014, The American journal of pathology.

[13]  Tom C. Freeman,et al.  Transcriptome-Based Network Analysis Reveals a Spectrum Model of Human Macrophage Activation , 2014, Immunity.

[14]  L. Nagy,et al.  Tissue LyC6− Macrophages Are Generated in the Absence of Circulating LyC6− Monocytes and Nur77 in a Model of Muscle Regeneration , 2013, The Journal of Immunology.

[15]  B. Chazaud,et al.  Monocyte/macrophage interactions with myogenic precursor cells during skeletal muscle regeneration , 2013, The FEBS journal.

[16]  B. Viollet,et al.  AMPKα1 regulates macrophage skewing at the time of resolution of inflammation during skeletal muscle regeneration. , 2013, Cell metabolism.

[17]  S. Moestrup,et al.  The Haptoglobin-CD163-Heme Oxygenase-1 Pathway for Hemoglobin Scavenging , 2013, Oxidative medicine and cellular longevity.

[18]  P. Taylor,et al.  Distinct bone marrow-derived and tissue resident macrophage-lineages proliferate at key stages during inflammation , 2013, Nature Communications.

[19]  K. Nakao,et al.  Secreted protein lipocalin‐2 promotes microglial M1 polarization , 2013, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[20]  A. Sica,et al.  Macrophage plasticity and polarization in tissue repair and remodelling , 2013, The Journal of pathology.

[21]  Ansuman T. Satpathy,et al.  Ly6C hi monocytes in the inflamed colon give rise to proinflammatory effector cells and migratory antigen-presenting cells. , 2012, Immunity.

[22]  Amin R. Mazloom,et al.  Gene-expression profiles and transcriptional regulatory pathways that underlie the identity and diversity of mouse tissue macrophages , 2012, Nature Immunology.

[23]  M. Rudnicki,et al.  Six1 regulates stem cell repair potential and self-renewal during skeletal muscle regeneration , 2012, The Journal of cell biology.

[24]  Daigo Hashimoto,et al.  Deciphering the transcriptional network of the DC lineage , 2012, Nature Immunology.

[25]  Charles R. Evans,et al.  The Sedoheptulose Kinase CARKL Directs Macrophage Polarization through Control of Glucose Metabolism , 2012, Cell metabolism.

[26]  Alberto Mantovani,et al.  Orchestration of metabolism by macrophages. , 2012, Cell metabolism.

[27]  Alberto Mantovani,et al.  Macrophage plasticity and polarization: in vivo veritas. , 2012, The Journal of clinical investigation.

[28]  Ruth C Lovering,et al.  Transcriptomic analyses of murine resolution-phase macrophages. , 2011, Blood.

[29]  P. Muñoz-Cánoves,et al.  p38/MKP-1–regulated AKT coordinates macrophage transitions and resolution of inflammation during tissue repair , 2011, The Journal of cell biology.

[30]  Carolyn L. Geczy,et al.  Inflammation-associated S100 proteins: new mechanisms that regulate function , 2011, Amino Acids.

[31]  Jennifer A. Lawson,et al.  Satellite cells , connective tissue fibroblasts and their interactions are crucial for muscle regeneration , 2022 .

[32]  T. Koh,et al.  Macrophage-Specific Expression of Urokinase-Type Plasminogen Activator Promotes Skeletal Muscle Regeneration , 2011, The Journal of Immunology.

[33]  A. Mantovani,et al.  Iron trafficking and metabolism in macrophages: contribution to the polarized phenotype. , 2011, Trends in immunology.

[34]  R. Ransohoff,et al.  Macrophages recruited via CCR2 produce insulin‐like growth factor‐1 to repair acute skeletal muscle injury , 2011, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[35]  P. Apostoli,et al.  Polarization dictates iron handling by inflammatory and alternatively activated macrophages , 2010, Haematologica.

[36]  T. Koh,et al.  Urokinase-type plasminogen activator increases hepatocyte growth factor activity required for skeletal muscle regeneration. , 2009, Blood.

[37]  F. Petrat,et al.  RELEASE OF REDOX-ACTIVE IRON BY MUSCLE CRUSH TRAUMA: NO LIBERATION INTO THE CIRCULATION , 2009, Shock.

[38]  Hiroyuki Aburatani,et al.  The S100A8–serum amyloid A3–TLR4 paracrine cascade establishes a pre-metastatic phase , 2008, Nature Cell Biology.

[39]  B. Pedersen,et al.  Calprotectin is released from human skeletal muscle tissue during exercise , 2008, The Journal of physiology.

[40]  N. Van Rooijen,et al.  Inflammatory monocytes recruited after skeletal muscle injury switch into antiinflammatory macrophages to support myogenesis , 2007, The Journal of experimental medicine.

[41]  G. Bassez,et al.  Muscle satellite cells and endothelial cells: close neighbors and privileged partners. , 2007, Molecular biology of the cell.

[42]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[43]  T. Braun,et al.  Mesenchymal stem cells are recruited to striated muscle by NFAT/IL-4-mediated cell fusion. , 2005, Genes & development.

[44]  F. Hanefeld,et al.  Monocyte/macrophage differentiation in dermatomyositis and polymyositis , 2004, Muscle & nerve.

[45]  Steffen Jung,et al.  Blood monocytes consist of two principal subsets with distinct migratory properties. , 2003, Immunity.

[46]  I. Kushner,et al.  Acute-phase proteins and other systemic responses to inflammation. , 1999, The New England journal of medicine.

[47]  N. Eriksen,et al.  Murine serum amyloid A3 is a high density apolipoprotein and is secreted by macrophages. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[48]  D. Harrison,et al.  Faculty of 1000 evaluation for Deciphering the transcriptional network of the dendritic cell lineage. , 2012 .

[49]  M. Albert,et al.  Muscle resident macrophages control the immune cell reaction in a mouse model of notexin-induced myoinjury. , 2010, Arthritis and rheumatism.