High fat diet induced hepatocyte exosomes-promoted hepatic inflammation and tumorigenesis

摘要 这项提案的总体目标是研究肝细胞外泌体在高脂饮食介导的肝细胞凋亡中的作用。 促进肝癌发展。此外，我们将确定是否使用新的可食用水果外泌体 作为用于将抗炎剂靶向递送至肿瘤细胞以治疗肝脏疾病的递送载体 癌我们的初步数据表明，肝细胞外泌体存在于外周血中， 肥胖小鼠和肥胖个体以及来自喂食高脂肪饮食（HFD-1）的小鼠的肝细胞外泌体。 外泌体）促进小鼠模型中的炎症反应和肝癌的生长。这些数据 肝细胞外泌体的产生与肝癌相关的炎症反应有关。 我们的初步数据表明，HFD-外泌体携带高水平的sumoylated CSN 5（sumo-CSN 5）和高水平的sumo-CSN 5。 向未成熟的骨髓细胞递送sumo-CSN 5促进了Jak 3的降解，这一过程 对骨髓细胞的分化至关重要。这指出，生产的sumo-csn 5作为一个关键的 外来体介导的炎症细胞因子诱导中的致病步骤。在肝细胞中，ErK 1/2激酶 被来自高脂喂养小鼠肝提取物上清液的可溶性因子激活， 作为CSN 5类小泛素化抑制剂的Senp 1的解离，从而促进CSN 5的产生 Sumo-CSN5这一过程被姜黄素抑制，姜黄素已知抑制ErK 1/2活化。因此，我们认为， 我们假设高脂饮食通过以下方式促进具有高水平sumo-CSN 5的外泌体的产生： 肝细胞;这些外来体被未成熟的骨髓细胞摄取，导致sumo-CSN 5- 通过增强Jak 3的降解介导免疫抑制，从而促进肿瘤生长。我们 进一步假设姜黄素治疗导致抑制CSN 5的类小泛素化的加工， 通过抑制ErK 1/2，从喂食高脂饮食的小鼠获得肝细胞。这些假设将在体外进行检验 （1）高脂肪肝细胞外泌体中的类小泛素化CSN 5对于肝细胞外泌体的形成是否是必需的。 激活的iMCs的积累和促进肝脏肿瘤进展;（2）ErK 1/2信号通路 决定了肝细胞中CSN 5的去小泛素化与小泛素化的平衡， 高脂饮食;和（3）高脂饮食诱导的肝细胞外来体介导的肝肿瘤生长促进， 通过使用新的可食用水果外来体作为用于靶向递送抗肿瘤药物的递送载体， 用于治疗癌症的炎性试剂与炎性细胞的结合。临床相关性。生成的数据 应该确定一种新的机制，将高脂肪饮食与炎症和免疫抑制反应联系起来 与肿瘤发展相关，从而确定化疗的分子靶点， 化学预防证明基于水果外泌体的姜黄素递送的安全性及其靶向 将是治疗这种使人衰弱的疾病的重要一步。

ABSTRACT The overall goal of this proposal is to study the role of hepatocyte exosomes in high fat diet mediated promotion of liver cancer development. Further, we will determine whether using novel edible fruit exosomes as a delivery vehicle for targetable delivery of an anti-inflammatory agent to tumor cells for treatment of liver cancer. Our preliminary data has indicated that hepatocyte exosomes are present in the peripheral blood of obese mice and obese individuals and the hepatocyte exosomes from mice fed a high-fat diet (HFD- exosomes) promote inflammatory responses and the growth of liver cancer in mouse model. These data implicate the production of hepatocyte exosomes in the inflammatory responses associated with liver cancer. Our preliminary data indicate that the HFD-exosomes carry high levels of sumoylated CSN5 (sumo-CSN5) and that the delivery of the sumo-CSN5 to immature myeloid promotes the degradation of Jak3, a process that is essential for differentiation of the myeloid cells. This pinpointed the production of the sumo-CSN5 as a critical pathogenic step in exosomes mediated Induction of inflammation cytokines. In hepatocyte cells, ErK1/2 kinase is activated by a soluble factor(s) from the supernatant of liver extracts of high fat-fed mice and causes the disassociation of Senp1 which acts as a suppressor of sumoylation of CSN5, thereby promoting the production of sumo-CSN5. This process is inhibited by curcumin, which is known to inhibit ErK1/2 activation. Therefore, we hypothesize that a high-fat diet promotes the production of exosomes with high levels of sumo-CSN5 by hepatocyte cells; that these exosomes are taken up by immature myeloid cells, resulting in sumo-CSN5- mediated immunosuppression by enhancing degradation of Jak3, and thus the promotion of tumor growth. We further hypothesize that curcumin treatment leads to the inhibition of processing of sumoylation of CSN5 in the hepatocyte cells from mice fed a high-fat diet by inhibition of ErK1/2. These hypotheses will be tested in vitro and in vivo to determine whether: (1) Sumoylated CSN5 in high-fat hepatocyte exosomes is essential for accumulation of activated iMCs and promotion of liver tumor progression; (2) The ErK1/2 signaling pathway determines the balance of desumoylation vs. sumoylation of CSN5 in hepatocyte cells and is promoted by a high-fat diet; and (3) the high-fat diet-induced hepatocyte exosome-mediated promotion of liver tumor growth is reversible by using novel edible fruit exosomes as a delivery vehicle for targetable delivery of an anti- inflammatory agent to inflammatory cells for treatment of cancer. Clinical Relevance. The data generated should identify a novel mechanism that links a high-fat diet to inflammatory and immunosuppressive responses associated with tumor development and thereby identify molecular targets for chemotherapy or chemoprevention. Demonstration of the safety of fruit exosomes-based delivery of curcumin and its targeting to inflammatory cells would be a significant step forward in the treatment of this debilitating disease.