Development of a new macrophage-specific TRAP mouse (MacTRAP) and definition of the renal macrophage translational signature

• Published in: Scientific reports Vol. 10; no. 1; p. 7519
• Main Authors: Hofmeister, Andreas, Thomaßen, Maximilian C., Markert, Sabrina, Marquardt, André, Preußner, Mathieu, Rußwurm, Martin, Schermuly, Ralph T., Steinhoff, Ulrich, Gröne, Hermann-Josef, Hoyer, Joachim, Humphreys, Benjamin D., Grgic, Ivica
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 05.05.2020; Nature Publishing Group
• Subjects: 38/35; 38/39; 38/91; 45/90; 631/114/2403; 64/110; 64/60; 692/308/575; 692/4022/272; 82/1; Animals; Clear cell-type renal cell carcinoma; Dendritic cells; Dendritic Cells - metabolism; Gene expression; Gene Expression Profiling; Genetic Techniques; Green Fluorescent Proteins - metabolism; Homeostasis; Humanities and Social Sciences; Immunohistochemistry; Inflammation; Kidney - cytology; Kidneys; Macrophages; Macrophages - metabolism; Mice; Mice, Transgenic; multidisciplinary; Protein Biosynthesis; Ribonucleic acid; Ribosomes - metabolism; RNA; RNA - metabolism; RNA-Seq; Science; Science (multidisciplinary); Transgenes; Transgenic mice; Translation
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-63514-6

Tissue macrophages play an important role in organ homeostasis, immunity and the pathogenesis of various inflammation-driven diseases. One major challenge has been to selectively study resident macrophages in highly heterogeneous organs such as kidney. To address this problem, we adopted a Translational Ribosome Affinity Purification (TRAP)- approach and designed a transgene that expresses an eGFP-tagged ribosomal protein (L10a) under the control of the macrophage-specific c-fms promoter to generate c-fms-eGFP-L10a transgenic mice (Mac TRAP ). Rigorous characterization found no gross abnormalities in Mac TRAP mice and confirmed transgene expression across various organs. Immunohistological analyses of Mac TRAP kidneys identified eGFP-L10a expressing cells in the tubulointerstitial compartment which stained positive for macrophage marker F4/80. Inflammatory challenge led to robust eGFP-L10a upregulation in kidney, confirming Mac TRAP responsiveness in vivo . We successfully extracted macrophage-specific polysomal RNA from Mac TRAP kidneys and conducted RNA sequencing followed by bioinformatical analyses, hereby establishing a comprehensive and unique in vivo gene expression and pathway signature of resident renal macrophages. In summary, we created, validated and applied a new, responsive macrophage-specific TRAP mouse line, defining the translational profile of renal macrophages and dendritic cells. This new tool may be of great value for the study of macrophage biology in different organs and various models of injury and disease.