Leir Shih-Hsing, Tkachenko Svyatoslav, Paranjapye Alekh, Van Wettere Arnaud J, Kerschner Jenny L, Perisse Iuri Viotti, Marriott Cheyenne M, Patrick Tayler, Liu Ying, White Kenneth L, Polejaeva Irina A, Harris Ann
Department of Genetics and Genome Sciences, Case Western Reserve University School of Medicine, Cleveland, OH, 44106-4955, USA.
Department of Genetics, University of Pennsylvania, Philadelphia, PA, USA.
Mol Med. 2025 Jun 11;31(1):231. doi: 10.1186/s10020-025-01266-7.
The Lung is the major focus of therapeutic approaches for the inherited disorder cystic fibrosis (CF) as without treatment lung disease is life-limiting. However, the initiating events that predispose the CF lung to cycles of infection, inflammation and resultant tissue damage are still unclear. Inflammation may occur in the CF lung prior to birth in human and several large animal models suggesting an in utero origin for the disease and encouraging further studies prior to birth.
Here we used the sheep model of CF (CFTR) and age-matched wild-type (WT) sheep of the same breed to investigate the single cell transcriptomes of proximal and distal lung tissue at 80 days and 120 days of gestation and at term (147 days). Single cell RNA-seq was performed on tissues from 4 to 7 animals of each genotype (WT and CFTR) at each time point.
At term, FOXJ1-expressing ciliated cells are overrepresented in both lung regions from CFTR lambs, while secretory epithelial and basal cells are underrepresented in proximal lung, as are T cells and monocytes in distal lung. The imbalance in ciliated and basal cells was confirmed by immunohistochemistry. At 120 days of gestation, lymphoid cells are slightly more abundant in proximal and distal lung from CFTR animals compared to WT, consistent with the transient CF-associated inflammatory response in utero. At 80 days of gestation, T and B cells are underrepresented in both lung regions.
The differences in epithelial cell abundance observed in the CFTR lambs at term may reflect sequelae from the loss of CFTR on lung development and differentiation in utero. These findings provide novel insights into the cellular mechanisms of pathology and may be relevant to the design of new therapeutic approaches for CF lung disease.
肺是遗传性疾病囊性纤维化（CF）治疗方法的主要关注焦点，因为未经治疗的肺部疾病会限制生命。然而，使CF肺易发生感染、炎症及由此导致的组织损伤循环的起始事件仍不清楚。在人类和几种大型动物模型中，CF肺在出生前可能就已发生炎症，这表明该疾病起源于子宫内，从而鼓励在出生前进行进一步研究。
在此，我们使用CF（CFTR）绵羊模型和同品种、年龄匹配的野生型（WT）绵羊，来研究妊娠80天、120天和足月（147天）时近端和远端肺组织的单细胞转录组。在每个时间点，对每种基因型（WT和CFTR）的4至7只动物的组织进行单细胞RNA测序。
足月时，CFTR羔羊的两个肺区域中表达FOXJ1的纤毛细胞比例过高，而近端肺中的分泌上皮细胞和基底细胞比例过低，远端肺中的T细胞和单核细胞比例也过低。免疫组织化学证实了纤毛细胞和基底细胞的失衡。与WT相比，妊娠120天时，CFTR动物近端和远端肺中的淋巴细胞略多，这与子宫内短暂的CF相关炎症反应一致。妊娠80天时，两个肺区域中的T细胞和B细胞比例过低。
足月时在CFTR羔羊中观察到的上皮细胞丰度差异可能反映了CFTR缺失对子宫内肺发育和分化的后遗症。这些发现为病理的细胞机制提供了新的见解，可能与CF肺部疾病新治疗方法的设计相关。
