Identifying a paracrine fibroblast-derived factor that stimulates fetal alveolar cells, thus potentially enhancing perinatal lung transition.

鉴定一种旁分泌成纤维细胞衍生因子，可刺激胎儿肺泡细胞，从而有可能增强围产期肺转变。

• 批准号: 454012436
• 负责人: Professor Dr. Ralf Hoffmann
• 金额: --
• 依托单位: Rudolf-Schönheimer-Institut für Biochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/454012436?language=en
• 关键词: Identifying; paracrine; fibroblast; derived; factor

Preterm infants represent the largest paediatric patient group and thus a large societal challenge. Lung immaturity in these infants is frequently associated with respiratory distress leading to significant morbidity and mortality. Currently, the treatment of infants with pulmonary complications is mainly symptomatic and rather unspecific with salient unwanted side effects. This situation highlights the urgent need to develop new therapeutic approaches aiming at a stimulation of lung maturation. Lung mesenchymal-derived paracrine cell signals stimulate alveolar functions like surfactant synthesis, but the contributing factors are largely unknown. In our previous work we determined a novel function of lung fibroblast-derived conditioned medium (LF-CM), the stimulation of epithelial sodium transport and ENaC gene expression in fetal alveolar cells. Vectorial sodium transport, a major function of lung epithelial cells, enables lung fluid absorption at birth and thus perinatal lung transition, which is impaired in preterm infants and possibly alleviated by LF-CM. In the proposed project our aim therefore is to identify the factor(s) responsible for the stimulation of sodium transport. The experimental setup will start with factor binding and separation by HPLC using a 4-dimensional separation scheme, followed by mass spectrometric factor identification and confirmation with synthetic compounds. The response of ENaC mRNA expression is tested and confirmed with every obtained LF-CM fraction as biological readout. Furthermore, it is planned to establish a faster, more robust readout, e.g. the Corning Epic System, to confirm the biological activity. The second strategy involves RNA-Sequencing comparing responder and non-responder cells to identify differentially regulated pathways by LF-CM. With these strategies, we expect to identify novel endogenous mesenchymal-derived factors able to stimulate lung maturation, and/or expand the knowledge about already known factors by determining additional previously unknown effects on lung epithelia. Furthermore, we want to characterize the involved pathways to identify new strategies to improve perinatal lung transition in preterm infants. Identifying these stimulating mesenchymal-derived paracrine components will enable clinical scientists in our group and elsewhere to develop urgently needed novel therapies with optimized effects and reduced side effects, thus improving the long term health of premature infants. Furthermore, these results may also open new therapeutic approaches for adult lung failure, including COVID-19 pneumonia, which also shows extreme distubances of alveolar fluid clearance. The joint effort of the Neonatology research group and the Institute of Bioanalytical Chemistry in cooperation with the Institute of Molecular Biochemistry and the Max Planck Institute für Heart- and Lung Research in Bad Nauheim will ensure successful execution of the project within the proposed time frame.

早产儿是最大的儿科患者群体，因此是一个巨大的社会挑战。这些婴儿的肺不成熟常常与呼吸窘迫相关，导致显着的发病率和死亡率。目前，对患有肺部并发症的婴儿的治疗主要是对症治疗，且特异性不强，具有明显的不良副作用。这种情况凸显了迫切需要开发旨在刺激肺成熟的新治疗方法。肺间充质来源的旁分泌细胞信号刺激肺泡功能，如表面活性剂合成，但其影响因素在很大程度上尚不清楚。在我们之前的工作中，我们确定了肺成纤维细胞来源的条件培养基 (LF-CM) 的新功能，即刺激胎儿肺泡细胞中上皮钠转运和 ENaC 基因表达。矢量钠转运是肺上皮细胞的一项主要功能，可促进出生时肺液的吸收，从而促进围产期肺的转变，早产儿的这种情况会受到损害，而 LF-CM 可能会减轻这种情况。因此，在拟议的项目中，我们的目标是确定刺激钠转运的因素。实验设置将从使用 4 维分离方案的 HPLC 进行因子结合和分离开始，然后使用合成化合物进行质谱因子鉴定和确认。使用每个获得的 LF-CM 级分作为生物读数来测试和确认 ENaC mRNA 表达的响应。此外，计划建立更快、更稳健的读数，例如Corning Epic System，以确认生物活性。第二种策略涉及 RNA 测序，比较应答细胞和非应答细胞，以识别 LF-CM 的差异调节途径。通过这些策略，我们期望识别能够刺激肺成熟的新型内源性间充质衍生因子，和/或通过确定对肺上皮的其他先前未知的影响来扩展关于已知因子的知识。 此外，我们希望描述所涉及的途径，以确定改善早产儿围产期肺转变的新策略。识别这些刺激性间充质来源的旁分泌成分将使我们小组和其他地方的临床科学家能够开发出急需的具有优化效果和减少副作用的新疗法，从而改善早产儿的长期健康。此外，这些结果还可能为成人肺衰竭（包括 COVID-19 肺炎）开辟新的治疗方法，这也显示出肺泡液清除的极大干扰。新生儿学研究小组和生物分析化学研究所与分子生物化学研究所和巴特瑙海姆马克斯·普朗克心肺研究所合作的共同努力将确保该项目在拟议的时间范围内成功执行。