MSC Exosome Treatment for BPD: Impact on Immunity and Lung Development

摘要 支气管肺发育不良(BPD)是早产儿的一种多因素慢性肺部疾病。没有单曲 预防或治疗BPD的有效治疗，需要新的治疗和降低风险的工具 与极端早产相关的进一步并发症的预防迫在眉睫。事实上，间充质基质/茎 细胞(MSC)疗法在BPD的临床前模型中显示出了希望，然而最近的研究表明 确定了MSCs的主要治疗载体包含在它们的分泌体中，并由 外显体。Exosome是大多数人表达的亚微米、脂双层包裹的细胞外小泡(EV)。 细胞。它们不同的起源、生物发生和分子组成使它们在不同的和强大的生理 角色，其中最耐人寻味的是一种细胞间通信的有效方法。MSC外切体 据报道，其组成包括小的非编码RNA、游离脂肪酸、表面受体和 蛋白质，作为MSC治疗效果的载体。因此，除了他们在其中的不同角色之外 对于健康和疾病，外切体是治疗应用的新型试剂。我们分离了外切体 来自人类MSC条件培养液，称为MEX，并表明它们抑制新生儿高氧中的BPD 老鼠模型。具体地说，一种剂量的MEX可以抑制肺部炎症、肺泡损伤、肺部 高血压，纤维化，并使长期肺功能正常化。我们已经证明墨西哥已经被占领了 通过巨噬细胞(MφS)，并因此将Mφ表型转变为消炎、抗纤维化和抗炎 改建。我们假设MφS是MEX治疗作用的关键载体，协调细胞到细胞 通讯信号，以促进正常的肺泡形成和恢复肺内平衡。我们将对此进行测试 以下具体目标中的假设：SA#1：分离并全面描述MEX 亚群并研究它们在体外和体内的作用和生物学效力的机制；SA#2：到 检测单核细胞/MφS作为MEX信号对肺细胞的介导者的作用；SA#3：研究其生物学特性 经MEX修饰的单核细胞/MφS在体内高氧诱导的BPD中的作用

ABSTRACT Bronchopulmonary dysplasia (BPD) is a multifactorial chronic lung disease of preterm infants. With no single effective therapy for either the prevention or treatment of BPD, the need for new tools to treat and reduce risk of further complications associated with extreme preterm birth is urgent. Indeed, mesenchymal stromal/stem cell (MSC) therapy had shown promise in preclinical models of BPD, however more recent studies have established that the main therapeutic vectors of MSCs is comprised in their secretome and represented by exosomes. Exosomes are submicron, lipid bilayer-enclosed extracellular vesicles (EVs) expressed by most cells. Their varied origin, biogenesis and molecular composition enroll them in diverse and potent physiological roles, the most intriguing of which is an effective method of cell-to-cell communication. The MSC exosome composition has been reported to include small noncoding RNAs, free fatty acids, surface receptors and proteins, serving as vectors of MSC therapeutic effects. Consequently, in addition to their diverse roles in health and disease, exosomes represent novel reagents for therapeutic applications. We isolated exosomes from human MSC conditioned media, termed MEx, and showed that they inhibit BPD in the neonatal hyperoxia mouse model. Specifically, one dose of Mex inhibits lung inflammation, alveolar injury, pulmonary hypertension, fibrosis, and normalizes long-term lung function. We have demonstrated that MEx are taken up by macrophages (Mφs) and, as result, shift the Mφ phenotype to inflammation resolving, antifibrotic, and anti- remodeling. We hypothesize that Mφs are key vectors of MEx therapeutic action, orchestrating cell-to-cell communication signals to promote normal alveogenesis and to restore lung homeostasis. We will test this hypothesis in the following specific aims: SA#1: To isolate and comprehensively characterize MEx subpopulations and investigate mechanisms of their action and biological potency in vitro and in vivo; SA#2: To test the role of monocytes/Mφs as mediators of MEx signals to lung cells; SA#3: To investigate the biologic function of monocytes/Mφs, modified by MEx, on hyperoxia-induced BPD in vivo.