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Interstitial macrophages (IMs), especially those derived from monocytes, play an unknown role in IPF pathogenesis. By using single-cell RNA sequencing (scRNA-seq), bleomycin (BLM)-induced pulmonary fibrosis mouse lungs were analyzed to characterize the cellular landscape and heterogeneity of macrophages in this model. scRNA-seq was used to identify distinct interstitial macrophage subpopulations in fibrotic lungs, with monocyte-derived macrophages exhibiting a pro-fibrotic gene expression profile enriched in wound healing, extracellular matrix (ECM) remodeling, and pro-fibrotic cytokine production functions. A pseudotime analysis revealed that IMs originated from monocytes and differentiated along a specific trajectory. A cell\u2013cell communication analysis demonstrated strong interactions between monocyte-derived interstitial macrophages (Mo-IMs) and fibroblasts through the transforming growth factor beta (TGF\u03b2), secreted phosphoprotein 1 (SPP1), and platelet-derived growth factor (PDGF) signaling pathways. Flow cytometry validated the presence and expansion of Mo-IMs subpopulations in BLM-treated mice. This study reveals the cellular heterogeneity and developmental trajectory of lung macrophages in early BLM-induced pulmonary fibrosis, highlighting the crucial role of Mo-IMs with a pro-fibrotic phenotype in IPF pathogenesis via interactions with fibroblasts. Targeting these specific macrophage subpopulations and associated signaling pathways may provide novel therapeutic strategies for IPF.<\/jats:p>","DOI":"10.3390\/ijms252111669","type":"journal-article","created":{"date-parts":[[2024,10,30]],"date-time":"2024-10-30T09:11:24Z","timestamp":1730279484000},"page":"11669","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Single-Cell RNA Sequencing Reveals Monocyte-Derived Interstitial Macrophages with a Pro-Fibrotic Phenotype in Bleomycin-Induced Pulmonary Fibrosis"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4473-4949","authenticated-orcid":false,"given":"Shunli","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caixia","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengyao","family":"Lei","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tong","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinxin","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suxia","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuting","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohan","family":"Bi","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fanshuo","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuyou","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0899-1270","authenticated-orcid":false,"given":"Ziling","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Pathology, Fudan University Shanghai Cancer Center, Shanghai 200032, China"},{"name":"Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianghua","family":"Yi","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1811","DOI":"10.1056\/NEJMra1705751","article-title":"Idiopathic Pulmonary Fibrosis","volume":"378","author":"Lederer","year":"2018","journal-title":"N. 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