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The role of resident lung mesenchymal stromal cells in the pathogenesis of bronchopulmonary dysplasia (BPD).

常驻肺间充质基质细胞在支气管肺发育不良（BPD）发病机制中的作用。

基本信息

批准号：
417891009
负责人：
Dr. Ivana Mizik, Ph.D.
金额：
--
依托单位：
Klinik und Poliklinik für Kinder- und Jugendmedizin
依托单位国家：
德国
项目类别：
Research Fellowships
财政年份：
2019
资助国家：
德国
起止时间：
2018-12-31 至 2020-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/417891009?language=en
关键词：
role resident lung mesenchymal stromal

项目摘要

Bronchopulmonary dysplasia (BPD), the chronic lung disease that follows ventilator and O2 therapy for acute respiratory failure, is the most common complication of prematurity and accounts for much of the long-term morbidity. BPD is characterized by impaired lung development and lacks effective therapies. Exogenous (bone marrow or umbilical cord) mesenchymal stromal cells (MSC) prevent O2-induced lung injury in newborn rodents. An apparent contradiction is the observation that premature babies are more likely to develop BPD if MSCs are present in their tracheal aspirates. Thus, in order to facilitate the clinical translation of this promising cell therapy, we will clarify the intriguing role of resident lung (L-)MSC in neonatal lung injury and repair.Hypothesis: The repair function of L MSCs is perturbed in experimental BPD and L-MSC dysfunction may contribute to the pathogenesis of BPD. Aim 1: To characterize L-MSCs in the normal developing mouse lung and determine if the function of L-MSCs is perturbed in experimental BPD. 1.1. We will isolate L-MSCs from normal mice and determine their characteristics (cell surface markers, differentiation and clonogenic potential and cytokine and extracellular matrix protein production). 1.2. We will use these same endpoints to compare L-MSCs from normal mice to mice with ventilation-induced lung injury (VILI) mimicking BPD. Mice will be exposed to LPS and mechanically ventilated for 8 hours. We will also perform RNAseq to compare the transcription of normal L-MSC compared to L-MSC isolated from mice with VILI. In addition, we will perform in vitro functional assays to compare the healing potential of L-MSCs. We will compare the effect of L-MSCs from control mice and from VILI mice on proliferation and wound healing of freshly isolated alveolar epithelial cells and proliferation and network formation on matrigel of lung endothelial cells.Aim 2: To determine if L-MSCs contribute to the pathogenesis of BPD. In vivo, we will inject normal mouse pups or pups exposed to hyperoxia (n=10/group) at P4 (onset of alveolar development) with either control L-MSCs (isolated previously from normally developing mice) or L-MSCs isolated from VILI mice and compare the effect on lung function and structure at P14. The strengths of this proposal include the novelty in exploring the role of endogenous L-MSCs, the readily availability of the animal models, well-established techniques to isolate and characterize L-MSCs, lung function and structure as well as the direct clinical implications. Indeed, the functional in vitro assays may serve as potency assays to predict the in vivo bioactivity of the cells. Furthermore, a better understanding of L-MSCs tissue specific function may contribute to develop superior exogenous MSC-based therapies. The relevance of our findings may extend to adult lung diseases characterized by alveolar damage such as emphysema and acute respiratory distress syndrome.

支气管肺发育不良(BPD)是呼吸机和氧气治疗急性呼吸衰竭后的慢性肺部疾病，是早产儿最常见的并发症，也是长期发病率的主要原因。BPD的特点是肺发育受损，缺乏有效的治疗方法。外源性(骨髓或脐带)间充质基质细胞(MSC)可预防O2诱导的新生啮齿动物肺损伤。一个明显的矛盾是观察到，如果早产儿的气管吸液中存在骨髓间充质干细胞，那么早产儿更有可能患上BPD。因此，为了便于这一有希望的细胞疗法的临床翻译，我们将阐明驻留肺(L-)间充质干细胞在新生儿肺损伤和修复中的有趣作用。假设：L骨髓间充质干细胞的修复功能在实验性BPD中受到干扰，L-MSC功能障碍可能参与了BPD的发病。目的1：鉴定L-间充质干细胞在正常发育小鼠肺中的特性，以确定L-间充质干细胞在实验性双侧肺损伤中的功能是否受到干扰。1.1.我们将从正常小鼠中分离L-MSCs，并对其特性(细胞表面标志、分化和克隆形成能力以及细胞因子和细胞外基质蛋白的产生)进行测定。1.2.我们将使用这些相同的终点来比较正常小鼠和模拟BPD的呼吸机诱导肺损伤(VILI)小鼠的L-MSCs。将小鼠暴露于脂多糖中，机械通风8小时。我们还将进行RNAseq，以比较正常的L-MSC和从VILI小鼠分离的L-MSC的转录。此外，我们还将进行体外功能分析，以比较L-MSCs的愈合潜力。我们将比较来自对照组和VILI小鼠的L-MSCs对新鲜分离的肺泡上皮细胞的增殖和创面愈合以及对肺内皮细胞基质的增殖和网络形成的影响。目的：探讨L-MSCs是否参与了BPD的发病机制。在体内，我们将正常小鼠或高氧暴露小鼠(n=10只/组)的幼鼠注射到P4(肺泡发育开始时)与对照组L-MSCs(先前从正常发育的小鼠分离)或从VILI小鼠分离的L-MSCs，并比较它们在P14对肺功能和结构的影响。这一建议的优点包括在探索内源性L-MSCs的作用方面的新颖性、动物模型的易得性、成熟的分离和鉴定L-MSCs的技术、肺功能和结构以及直接的临床意义。事实上，体外功能分析可以作为预测细胞体内生物活性的效力分析。此外，更好地了解L-MSCs的组织特异性功能可能有助于开发更好的基于外源性MSC的疗法。我们的发现可能延伸到以肺泡损伤为特征的成人肺部疾病，如肺气肿和急性呼吸窘迫综合征。