Plant exosomes non-coding RNA-mediated anti-inflammatory mechanisms

 描述（由申请人提供）：本提案的总体目标是使用葡聚糖硫酸钠（DSS）诱导的结肠炎和其他三种肠道炎症相关小鼠模型，确定可食用植物外泌体非编码小RNA（nc-sRNA）在预防肠道炎症中的作用。虽然有证据支持大多数可食用植物作为饮食的一部分的有益效果，但特定植物性补充剂有益的证据存在争议，其潜在机制尚不清楚。挑战在于分离和表征特定天然产物中所含的基本功能单元的生物活性，以确定其对宿主的细胞和分子效应是否以及如何以有益的方式使用。我们最近发表的数据（1-5）表明，存在于许多可食用植物（包括葡萄柚和生姜）中的外泌体样纳米颗粒（ELN）具有抗炎特性。喂食葡萄柚ELN（FELN）的小鼠受到保护免于DSS诱导的小鼠结肠炎。肠巨噬细胞摄取FELN抑制IL-1β和IL-6的释放以及HO-1和IL-10表达的诱导。我们的初步数据进一步表明，FELN携带高水平的特异性nc-sRNA，通过靶向肠道巨噬细胞抑制促炎细胞因子的诱导。nc-sRNA与巨噬细胞miR 155的失活相关。与柚苷复合的特异性FELN nc-sRNA与miR 155结合，随后阻止miR 155与某些抗炎基因的3-UTR结合，从而抑制促炎细胞因子的诱导。我们还证明了生姜ELN nc-sRNAs在稳态以及炎症条件下促进固有层巨噬细胞中IL-10的产生。因此，我们假设携带nc-sRNA生物活性复合物的植物ELN被肠道巨噬细胞摄取，随后与炎性microRNA结合，从而抑制慢性炎症细胞因子的诱导，从而预防慢性肠道炎症。我们的假设将在体外和体内进行测试以确定：（1）肠巨噬细胞摄取的FELN是否通过nc-sRNA-155介导的抗炎作用导致耐受性树突状细胞（DC）的诱导，因为DC通过与肠巨噬细胞的串扰对于肠免疫耐受和稳态是至关重要的;（2）调节抗炎nc-sRNA-155分选到巨噬细胞外泌体中的因子;和（3）哪些姜ELN nc-sRNA是诱导巨噬细胞中表达的抗炎IL-10所需的。ELN复合物靶向肠道炎症细胞的证明将是理解植物界如何与哺乳动物物种相互作用以通过ELN调节抗炎反应的重要一步。所产生的数据应该为从不同类型的植物中选择特定的ELN为患者提供个性化的补充药物提供基础，并确定口服施用从不同植物分离的定制外泌体是否会对疾病的预防或治疗具有协同/累加作用。

 DESCRIPTION (provided by applicant): The overall goal of this proposal is to define the role of edible plant exosomal non-coding small RNA (nc-sRNA) in the prevention of gut inflammation using the dextran sulfate sodium (DSS) induced colitis and three other gut inflammatory related mouse models. Although evidence is supportive of the beneficial effects for most edible plants taken as part of the diet, the evidence that specific plant-based supplements are beneficial is controversial, and the underlying mechanisms are not clear. The challenge is to isolate and characterize the biological activities of the essential functional unit contained in specific naturl products to determine if and how it's cellular and molecular effects on the host can be used in a beneficial manner. Our recently published data (1-5) indicates that exosome-like nanoparticles (ELNs) present in a number of edible plants including grapefruit and ginger have anti-inflammatory properties. Mice fed grapefruit ELNs (FELNs) are protected against DSS induced mouse colitis. Up take of FELNs by intestinal macrophages inhibits the release of IL-1β and IL-6 and the induction of HO-1 and IL-10 expression. Our preliminary data further indicate that FELNs carry high levels of specific nc-sRNA that inhibit the induction of proinflammatory cytokines by targeting intestinal macrophages. The nc-sRNA is associated with inactivation of macrophage miR155. A specific FELN nc-sRNA complexed with naringin binds to miR155 and subsequently prevents miR155 binding to 3-UTRs of certain anti-inflammatory genes, thus inhibiting the induction of proinflammatory cytokines. We have also demonstrated that ginger ELN nc-sRNAs promotes the production of IL-10 in lamina propria macrophages under steady-state as well as inflammatory conditions. Therefore, we hypothesize that plant ELNs carrying the nc-sRNA bioactive complex are taken up by intestinal macrophages and subsequently bind to inflammatory microRNAs, resulting in inhibition of induction of chronic inflammation cytokines, thereby preventing chronic intestinal inflammation. Our hypothesis will be tested in vitro and in vivo to determine: (1) whether FELN taken up by intestinal macrophages leads to induction of tolerant dendritic cells (DCs) through nc-sRNA-155 mediated anti-inflammatory effects since DCs are critical for gut immune tolerance and homeostasis through cross-talk with intestinal macrophages; (2) the factors that regulate sorting of anti-inflammatory nc-sRNA-155 into macrophage exosomes; and (3) which ginger ELNs nc-sRNA(s) are required for induction of anti- inflammatory IL-10 expressed in macrophages. Demonstration of the ELNs complex targeting to gut inflammatory cells would be a significant step forward in the understanding of how the plant kingdom interacts with the mammalian species to regulate anti-inflammatory responses through ELNs. The data generated should provide a foundation for selecting specific ELNs from different types of plants for personalized complementary medicine for patients and determine whether oral administration of a customized exosome isolated from different plants will have a synergistic/additive effect on prevention or treatment of disease.