以lncRNA为靶点的iPSC来源间充质干细胞对变应性鼻炎ILC2的免疫调控

Currently allergic rhinitis (AR) has a high incidance. Conventionally, AR inflammation is characterized as allergen-specific Th2 response. However, group 2 innate lymphoid cells (ILC2) was recently reported to be initated by the epithelial cell-derived cytokines and play the key role in the asthma inflammation via producing a lot of canonical type 2 cytokines IL-5 and IL-13.However, there were different reports about ILC2 levels in AR. We found that ILC2 levels significantly increased in AR and had a positive correlation with the severity of the diseases. We firstly reported that mesenchymal stem cells (MSC)derived from induced pluripotent stem cells (iPSC) prevented allergic rhinitis inflammation and down-regulated the function of Th2 cells. We did some pre-experiments and found that the levels of ILC2 in AR patients and AR mice were decreaed after iPSC-MSC treatment. However, it should be further studied for the details. Furthermore, the mechnisms that the effects of iPSC-MSC on ILC2 are unclear.Long non-coding RNA (lncRNA) was reported to be involved in the development and the function of T cells. We cultured ILC2 from AR patients with iPSC-MSC and did a lncRNA microarray. The lncRNAs with different expression were identified and few lncRNAs were further selected after deep data analysis and the confirmation with QT-PCR. After interfering lncRNA function, we found that iPSC-MSC regulated ILC2 function via lncRNA. Then this project will further study the effects of iPSC-MSC on ILC2 using AR patient PBMC and in allergic airway inflammation mice. With the technique of transfection, we will study the role of lncRNAs in AR inflammation and whether iPSC-MSC play their immunomodulation on ILC2 by targeting lncRNA in AR patients and mouse model of allergic airway inflammation. This projects will provide useful information to the mechnism of the immunomodulation of iPSC-MSC on ILC2.

变应性鼻炎(AR)为常见病，最近报道II 型固有淋巴细胞（ILC2）在哮喘的发病中起着和Th2细胞同等的作用，但AR的结果报道不一。我们前期发现AR患者ILC2水平升高，并报道了iPSC来源的间充质干细胞（MSC）可有效抑制AR炎症，预实验证明其可下调AR患者及小鼠ILC2水平，但尚需深入的研究，另外iPSC-MSC对ILC2的作用机制亦需进一步探讨。近几年报道长链非编码RNA(LncRNA)可影响T细胞功能，我们前期将AR患者ILC2与iPSC-MSC共培养并应用芯片筛选出差异表达的lncRNA且进一步锁定了候选转录本，通过转染初步提示iPSC-MSC可通过lncRNA调节ILC2功能。本课题拟应用AR患者标本及模型小鼠深入探讨iPSC-MSC对ILC2的作用，并探讨lncRNA在AR发病中的作用及是否是iPSC-MSC调节ILC2的靶点，将为iPSC-MSC的免疫调节提供理论依据。

变应性鼻炎（AR）及哮喘是全球公共健康问题。近期发现ILC2可以大量分泌2型细胞因子。我们前期报道了iPSC来源的间充质干细胞（MSC）可有效抑制呼吸道变应性炎症，因此具有调节ILC2功能潜力。lncRNA可调节DC的分化和Treg细胞的功能，表明可能与气道过敏性炎症有关。我们假设ILC2在呼吸道变应性炎症中起重要作用；MSC可通过调控ILC2而抑制过敏炎症且机制与lncRNA相关。本项目①应用AR及哮喘患者临床标本及动物模型探讨ILC2及其与DC、上皮细胞的相互作用在呼吸道变应性炎症发病中的作用;②探讨iPSC-MSC及其外泌体对呼吸道变应性炎症ILC2等的免疫调节作用;③以lncRNA、miRNA等为靶点的iPSC-MSC免疫调节作用的机制。结果：①阐明了AR及哮喘患者外周血中ILC2的水平升高，激素治疗后水平均出现显著下降；证明了髓系DC可以通过IL-33/ST2通路促进AR患者ILC2的活化，而浆系DC则可通过IL-6抑制ILC2功能; 发现AR患者外周血ILC2表面有白三烯受体表；证明了ILC2存在性别差异。②证明了MSC可通过激活Treg 细胞而抑制AR患者ILC2的功能；证明MSC外泌体可抑制AR患者及小鼠模型ILC2的功能; 证明了MSC外泌体可调控AR巨噬细胞功能；阐明了MSC可通过线粒体转移、miR-21/ ACVR2A 水平而调控上皮细胞的功能。③阐明了lncRNA 参与iPSC-MSC 对小鼠呼吸道变应性炎症的免疫调节，并筛选出了目标lncRNA；证明了MSC外泌体可通过miR-146a-5p而调节呼吸道变应性炎症的ILC2功能。本项目的实施完善了以ILC2为中心的呼吸道变应性炎症的发病机制，提供MSC及其外泌体这一新型生物治疗方案，为AR的防治提供了理论基础，具有较好的转化应用前景。依托本项目共发表论文19篇，培养研究生9名。