Novel strategies for therapeutic programming and delivery of mesenchymal stem cells to improve outcome of influenza virus-induced lung injury

治疗编程和间充质干细胞递送的新策略可改善流感病毒引起的肺损伤的结果

• 批准号: 319864050
• 负责人: Professor Dr. Thomas Braun
• 金额: --
• 依托单位: Medizinische Klinik V, Klinik für Infektiologie und Krankenhaushygiene
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/319864050?language=en
• 关键词: Novel; strategies; therapeutic; programming; delivery

Mesenchymal stem cells (MSC) have promising therapeutic potential in different forms of acute lung injury (ALI), and are now in first clinical trials. However, the molecular mechanisms underlying injury- or pathogen-related beneficial effects within defined niches of the affected lung are mostly unknown. Likewise, the mechanisms by which therapeutically applied MSC of different sources, such as bone marrow or adipose tissue, and lung-resident MSC (rMSC) sense injury during viral lung infections have not been comprehensively examined. This project focuses on the functional heterogeneity of MSC and characterizes the molecular interactions between virus-infected lung cells and different subsets of MSC with dedicated anti-viral and organ repair capacity. In the first funding period, we used bulk- and single cell (sc)RNA-Seq analyses combined with advanced lung organoid modelling and in vivo infection models to demonstrate that bone marrow-derived MSC (BM-MSC), white adipose tissue-derived MSCs (WAT-MSC) and rMSC represent heterogeneous populations that differ in single cell transcriptional profiles, exerting distinct roles in anti-viral host defense, tissue protection, and lung tissue regeneration (re-alveolarization). MSC of different origin could be primed in an injury-specific way to effectively support (re)generation of the alveolar compartment, e.g. by IV-specific preconditioning and by TLR3 ligands ex vivo, and in IV- and LPS-injury models in vivo. Regarding the potent antiviral and tissue-protective effects of MSC, we identified a type I IFN-dependent cross-talk as the underlying signalling event between BM-MSC and the IV-infected AEC, both in in vitro models and after intrapulmonary delivery of BM-MSC into IV-infected WT or IFNAR-/- mice in vivo.In the upcoming funding period, we will (i) molecularly and functionally characterize MSC clusters defined by distinct gene expression profiles, which are highly supportive for (re)alveolarization in lung organoid models and for alveolar repair in in vivo viral infection models. We will determine whether e.g. SPARC, SERPINF1, IGF1 and CSF1/2 identified to be upregulated in injured lung MSC in our RNA-seq analyses account - among others - for the regenerative potential of MSC, or might even substitute for MSC supernatants. To investigate further candidate molecules; we will (ii) define the functional relevance of TLR-responses induced in MSC during IV infection and/or LPS challenge using gain- and loss-of-function approaches in vivo in lung injury models; and (iii) validate our findings in human GCP-produced clinical-grade MSC in human ex vivo models, applying strategies such as disease-specific pre-conditioning, gene transfer, and genome editing to endow human MSC with defined genetic programs and beneficial factors to ultimately use them in first-in-human trials.

间充质干细胞（MSC）在不同形式的急性肺损伤（ALI）中具有良好的治疗潜力，目前正在进行首次临床试验。然而，在受影响肺的特定生态位中，与损伤或病原体相关的有益作用的分子机制大多是未知的。同样，用于治疗的不同来源的间充质干细胞，如骨髓或脂肪组织，以及病毒性肺部感染期间肺内间充质干细胞（rMSC）感觉损伤的机制尚未得到全面研究。本项目侧重于MSC的功能异质性，并表征病毒感染的肺细胞与具有专用抗病毒和器官修复能力的MSC不同亚群之间的分子相互作用。在第一个资助期，我们使用大细胞和单细胞（sc）RNA-Seq分析结合先进的肺类器官建模和体内感染模型来证明骨髓来源的间充质干细胞（BM-MSC），白色脂肪组织来源的间充质干细胞（watt -MSC）和rMSC代表异质群体，其单细胞转录谱不同，在抗病毒宿主防御，组织保护和肺组织再生（再肺泡化）中发挥不同的作用。不同来源的间充质干细胞可以通过损伤特异性的方式激活，从而有效地支持肺泡间室的（再）生成，例如通过体外的IV特异性预处理和TLR3配体，以及体内的IV和lps损伤模型。关于MSC的强效抗病毒和组织保护作用，我们在体外模型和体内将bmp -MSC肺内输送到iv感染的WT或IFNAR-/-小鼠后，发现了I型ifn依赖的串扰作为bmp -MSC与iv感染的AEC之间潜在的信号事件。在即将到来的资助期内，我们将(i)从分子和功能上表征由不同基因表达谱定义的MSC集群，这高度支持肺类器官模型中的（再）肺泡化和体内病毒感染模型中的肺泡修复。在我们的RNA-seq分析中，我们将确定在受损肺间充质干细胞中被鉴定为上调的SPARC、serinf1、IGF1和CSF1/2是否能够解释MSC的再生潜力，或者甚至可能替代MSC上清液。进一步研究候选分子；我们将（ii）在肺损伤模型中使用获得和丧失功能的方法，定义在IV感染和/或LPS攻击期间，MSC诱导的tlr反应的功能相关性；（iii）在人类离体模型中验证我们在人类gcp生产的临床级MSC中的发现，应用疾病特异性预处理、基因转移和基因组编辑等策略，赋予人类MSC明确的遗传程序和有益因素，最终将其用于首次人体试验。