@article{11b2035a4ca14b32a27649af52ace1eb,
title = "A Cre-deleter specific for embryo-derived brain macrophages reveals distinct features of microglia and border macrophages",
abstract = "Genetic tools to target microglia specifically and efficiently from the early stages of embryonic development are lacking. We generated a constitutive Cre line controlled by the microglia signature gene Crybb1 that produced nearly complete recombination in embryonic brain macrophages (microglia and border-associated macrophages [BAMs]) by the perinatal period, with limited recombination in peripheral myeloid cells. Using this tool in combination with Flt3-Cre lineage tracer, single-cell RNA-sequencing analysis, and confocal imaging, we resolved embryonic-derived versus monocyte-derived BAMs in the mouse cortex. Deletion of the transcription factor SMAD4 in microglia and embryonic-derived BAMs using Crybb1-Cre caused a developmental arrest of microglia, which instead acquired a BAM specification signature. By contrast, the development of genuine BAMs remained unaffected. Our results reveal that SMAD4 drives a transcriptional and epigenetic program that is indispensable for the commitment of brain macrophages to the microglia fate and highlight Crybb1-Cre as a tool for targeting embryonic brain macrophages.",
keywords = "BAMs, SMAD4, fate-mapping, macrophages ontogeny, microglia",
author = "Simone Brioschi and Belk, {Julia A.} and Vincent Peng and Martina Molgora and Rodrigues, {Patrick Fernandes} and Nguyen, {Khai M.} and Shoutang Wang and Siling Du and Wang, {Wei Le} and Grajales-Reyes, {Gary E.} and Ponce, {Jennifer M.} and Yuede, {Carla M.} and Qingyun Li and Baer, {John M.} and DeNardo, {David G.} and Susan Gilfillan and Marina Cella and Satpathy, {Ansuman T.} and Marco Colonna",
note = "Funding Information: We thank Dr. E. Lantelme for the excellent management of the flow cytometry core facility at the Pathology and Immunology Department (Washington University in St. Louis). We thank Dr. W. Beatty from the Molecular Microbiology Department (Washington University in St. Louis) for the excellent management of the imaging core. We thank Drs. M. Blurton-Jones (University of California Irvine) and D. Hume (University of Queensland) for providing the Fire knockout mice; Dr. K. Murphy (Washington University in St. Louis) for providing the Zbtb46GFP mice; and B. Bhattarai for sharing the Smad4F/F mice. We also thank Dr. Y. Zhou for helping set up the nuclei extraction protocol, J. Sun for helping collect preliminary data, A. Swain (10x Genomics) for helpful suggestions regarding the multiomics protocol, and Richard Cho (Cell Signaling) for the generous supply of antibodies. Lastly, we thank the Genome Engineering and iPSC Center (GEiC) (Washington University in St. Louis) for gRNA validation services, as well as Mike White and the Transgenic, Knockout, and Micro-Injection Core at the Pathology and Immunology Department (Washington University in St. Louis) for generating the genetically modified mice used in this study. M. Colonna is supported by the Rheumatic Diseases Research Resource-Based Center at Washington University (NIH/NIAMS P30AR073752). The Animal Behavior Core is supported in part by funding from the McDonnell Centers for Cellular and Molecular and Systems Neuroscience. This work was supported by NIH RF1AG05148501, RF1AG059082, R21AG059176, R01AI158579, and Cure Alzheimer's Fund (to M. Colonna). A.T.S. is supported by a Career Award for Medical Scientists from the Burroughs Wellcome Fund and a Pew-Stewart Scholars Award. J.A.B. is supported by a Stanford Graduate Fellowship and the National Science Foundation Graduate Research Fellowship under grant no. DGE-1656518. Cartoons and graphical abstract were created with BioRender.com. Conceptualization: S.B.; methodology, S.B. J.A.B. M.M. P.F.R. K.M.N. S.W. W.-L.W. C.M.Y. and Q.L.; software: J.A.B. and V.P.; validation: S.B. and S.D.; formal analysis: S.B. J.A.B. and V.P.; investigation: S.B. and J.A.B.; resources: G.G.-R. D.D. J.B. J.P. S.G. and M. Cella; data curation: S.B. J.A.B. and V.P.; writing manuscript: S.B.; review and editing: M.M. P.F.R. J.A.B. A.T.S. and M. Colonna; funding acquisition: S.B. and M. Colonna; supervision: M. Colonna and A.T.S. A.T.S. is a founder of Immunai and Cartography Biosciences and receives research funding from Allogene Therapeutics and Merck Research Laboratories. Funding Information: A.T.S. is a founder of Immunai and Cartography Biosciences and receives research funding from Allogene Therapeutics and Merck Research Laboratories. Funding Information: We thank Dr. E. Lantelme for the excellent management of the flow cytometry core facility at the Pathology and Immunology Department (Washington University in St. Louis). We thank Dr. W. Beatty from the Molecular Microbiology Department (Washington University in St. Louis) for the excellent management of the imaging core. We thank Drs. M. Blurton-Jones (University of California Irvine) and D. Hume (University of Queensland) for providing the Fire knockout mice; Dr. K. Murphy (Washington University in St. Louis) for providing the Zbtb46 GFP mice; and B. Bhattarai for sharing the Smad4 F/F mice. We also thank Dr. Y. Zhou for helping set up the nuclei extraction protocol, J. Sun for helping collect preliminary data, A. Swain (10x Genomics) for helpful suggestions regarding the multiomics protocol, and Richard Cho (Cell Signaling) for the generous supply of antibodies. Lastly, we thank the Genome Engineering and iPSC Center (GEiC) (Washington University in St. Louis) for gRNA validation services, as well as Mike White and the Transgenic, Knockout, and Micro-Injection Core at the Pathology and Immunology Department (Washington University in St. Louis) for generating the genetically modified mice used in this study. M. Colonna is supported by the Rheumatic Diseases Research Resource-Based Center at Washington University ( NIH / NIAMS P30AR073752). The Animal Behavior Core is supported in part by funding from the McDonnell Centers for Cellular and Molecular and Systems Neuroscience. This work was supported by NIH RF1AG05148501, RF1AG059082, R21AG059176, R01AI158579, and Cure Alzheimer{\textquoteright}s Fund (to M. Colonna). A.T.S. is supported by a Career Award for Medical Scientists from the Burroughs Wellcome Fund and a Pew-Stewart Scholars Award. J.A.B. is supported by a Stanford Graduate Fellowship and the National Science Foundation Graduate Research Fellowship under grant no. DGE-1656518. Cartoons and graphical abstract were created with BioRender.com . Publisher Copyright: {\textcopyright} 2023 Elsevier Inc.",
year = "2023",
month = may,
day = "9",
doi = "10.1016/j.immuni.2023.01.028",
language = "English",
volume = "56",
pages = "1027--1045.e8",
journal = "Immunity",
issn = "1074-7613",
number = "5",
}