Cantu Abiud, Gutierrez Manuel Cantu, Dong Xiaoyu, Leek Connor, Anguera Montserrat, Lingappan Krithika
bioRxiv. 2023 Aug 13:2023.08.09.552532. doi: 10.1101/2023.08.09.552532.
Recovery from lung injury during the neonatal period requires the orchestration of many biological pathways. The modulation of such pathways can drive the developing lung towards proper repair or persistent maldevelopment that can lead to a disease phenotype. Sex as a biological variable can regulate these pathways differently in the male and female lung exposed to neonatal hyperoxia. In this study, we assessed the contribution of cellular diversity in the male and female neonatal lung following injury. Our objective was to investigate sex and cell-type specific transcriptional changes that drive repair or persistent injury in the neonatal lung and delineate the alterations in the immune-endothelial cell communication networks using single cell RNA sequencing (sc-RNAseq) in a murine model of hyperoxic injury. We generated transcriptional profiles of >55,000 cells isolated from the lungs of postnatal day 1 (PND 1) and postnatal day 21 (PND 21) neonatal male and female C57BL/6 mice exposed to 95% FiO between PND 1-5 (saccular stage of lung development). We show the presence of sex-based differences in the transcriptional states of lung endothelial and immune cells at PND 1 and PND 21. Furthermore, we demonstrate that biological sex significantly influences the response to injury, with a greater number of differentially expressed genes showing sex-specific patterns than those shared between male and female lungs. Pseudotime trajectory analysis highlighted genes needed for lung development that were altered by hyperoxia. Finally, we show intercellular communication between endothelial and immune cells at saccular and alveolar stages of lung development with sex-based biases in the crosstalk and identify novel ligand-receptor pairs. Our findings provide valuable insights into the cell diversity, transcriptional state, developmental trajectory, and cell-cell communication underlying neonatal lung injury, with implications for understanding lung development and possible therapeutic interventions while highlighting the crucial role of sex as a biological variable.
新生儿期肺损伤的恢复需要多种生物学途径的协调作用。这些途径的调节可促使发育中的肺朝着正常修复或持续发育异常的方向发展，而持续的发育异常可能导致疾病表型。性别作为一种生物学变量，在暴露于新生儿高氧环境的雄性和雌性肺中对这些途径的调节可能有所不同。在本研究中，我们评估了损伤后雄性和雌性新生儿肺中细胞多样性的作用。我们的目的是研究性别和细胞类型特异性的转录变化，这些变化驱动新生儿肺的修复或持续损伤，并使用单细胞RNA测序（sc-RNAseq）在高氧损伤小鼠模型中描绘免疫内皮细胞通讯网络的改变。我们生成了从出生后第1天（PND 1）和出生后第21天（PND 21）的新生儿雄性和雌性C57BL/6小鼠肺中分离出的超过55,000个细胞的转录谱，这些小鼠在PND 1至5期间（肺发育的囊状期）暴露于95%的FiO₂。我们显示在PND 1和PND 21时，肺内皮细胞和免疫细胞的转录状态存在基于性别的差异。此外，我们证明生物性别显著影响对损伤的反应，与雄性和雌性肺共有的差异表达基因相比，更多的差异表达基因呈现出性别特异性模式。伪时间轨迹分析突出了高氧改变的肺发育所需基因。最后，我们展示了肺发育囊状期和肺泡期内皮细胞与免疫细胞之间的细胞间通讯，在相互作用中存在基于性别的偏向，并鉴定了新的配体-受体对。我们的研究结果为新生儿肺损伤背后的细胞多样性、转录状态、发育轨迹和细胞间通讯提供了有价值的见解，对理解肺发育和可能的治疗干预具有启示意义，同时突出了性别作为生物学变量的关键作用。
