Wu Shuang, Drive Ian, Luo Meihui, Miyazaki Hikaru, Shambhu Smitha, Popov Dimitry, Yang Liyuan, Wang Jing, Ma Jia, Guo Junfeng, Atha Jarron, Tata Aleksandra, Hoffman Eric A, Wang Yujiong, Tata Purushothama Rao, Jensen Martin B, Engelhardt John F, Sontake Vishwaraj, Liu Xiaoming
Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in Western China, College of Life Science, Ningxia University, Yinchuan, Ningxia 750021, PR China.
bioRxiv. 2025 May 14:2025.05.08.652970. doi: 10.1101/2025.05.08.652970.
Idiopathic pulmonary fibrosis (IPF) is a debilitating lung disease with limited therapeutic options. The development of effective therapies has been hindered by the lack of models that recapitulate key features of human disease. Here we report a bleomycin-induced ferret PF model characterized by an irreversible decrease in pulmonary compliance and an increase of opacification, accompanied by "honeycomb cyst-like" structures and "proximalization" of distal lung epithelium. Cellular and molecular analysis by single-nucleus RNA sequencing revealed a significant shift in distal lung epithelium towards proximal epithelial phenotype. Importantly, a histopathological pattern of bronchiolization encompassing divergent atypical epithelial cells and KRT17/TP63/KRT5 "basaloid-like" cells was present in the distal fibrotic lung lesions. Trajectory analysis revealed AT2 cells transition through multiple cell-states in bleomycin injured ferret lungs, particularly AT2 to KRT8/KRT7/SOX4 and eventually to KRT8/KRT7/SFN/TP63/KRT5 "basaloid-like" cells. Further, immunofluorescence analyses demonstrated KRT7 and KRT8 populations reside overlaying the ACTA2-positive myofibroblasts in fibrotic foci, implying their pro-fibrogenic activity similar to human IPF lungs. Collectively, our results provide evidence that bleomycin-induced lung injury in ferrets recapitulates pathophysiological, cellular, and molecular features of human IPF, suggesting that they may be a reliable model for understanding mechanisms of IPF pathogenesis and for testing therapeutic strategies for treatment of IPF.
特发性肺纤维化（IPF）是一种使人衰弱的肺部疾病，治疗选择有限。由于缺乏能够重现人类疾病关键特征的模型，有效疗法的开发受到了阻碍。在此，我们报告了一种博来霉素诱导的雪貂肺纤维化模型，其特征为肺顺应性不可逆降低和肺混浊增加，伴有“蜂窝状囊肿样”结构以及远端肺上皮的“近端化”。通过单核RNA测序进行的细胞和分子分析显示，远端肺上皮向近端上皮表型发生了显著转变。重要的是，在远端纤维化肺病变中存在一种细支气管化生的组织病理学模式，包括不同的非典型上皮细胞和KRT17/TP63/KRT5“基底样”细胞。轨迹分析显示，在博来霉素损伤的雪貂肺中，II型肺泡上皮细胞（AT2）经历多种细胞状态转变，特别是从AT2转变为KRT8/KRT7/SOX4，最终转变为KRT8/KRT7/SFN/TP63/KRT5“基底样”细胞。此外，免疫荧光分析表明，KRT7和KRT8细胞群体位于纤维化病灶中α-平滑肌肌动蛋白（ACTA2）阳性肌成纤维细胞之上，这意味着它们具有与人类IPF肺相似的促纤维化活性。总体而言，我们的结果提供了证据，表明博来霉素诱导的雪貂肺损伤重现了人类IPF的病理生理、细胞和分子特征，这表明它们可能是理解IPF发病机制以及测试IPF治疗策略的可靠模型。
