Amelioration of Doxorubicin Induced Muscle Dysfunction with Embryoinic stem cells-Derived Exosomes

利用胚胎干细胞来源的外泌体改善阿霉素引起的肌肉功能障碍

• 批准号: 10322656
• 负责人: Rakesh C Kukreja
• 金额: $ 49万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322656
• 关键词: Anti-Inflammatory Agents; Antineoplastic Agents; Apoptosis; Attenuated; C57BL/6 Mouse; CASP1 gene; Cancer Patient; Cell Death; Cell Death Induction; Cells; Clinic; Data; Development; Disease; Doxorubicin; Drug Side Effects; Fatigue; Functional disorder; Grant; Human; IL18 gene; Immune; In Vitro; Infiltration; Inflammasome; Inflammation; Inflammatory; Interleukin-1 beta; Knockout Mice; MAPK8 gene; Macrophage; Malignant Neoplasms; Mediating; MicroRNAs; Mus; Muscle; Muscle Fatigue; Muscle Weakness; Muscle function; Muscular Atrophy; Myocardium; Necrosis; Organ; Pathogenesis; Pathway interactions; Patient-Focused Outcomes; Patients; Phosphorylation; Preparation; Prevalence; Property; Reporting; Role; Signal Transduction; Skeletal Muscle; Small Interfering RNA; Soleus Muscle; Source; TLR4 gene; Teratoma; Testing; Therapeutic; Tissues; Toll-like receptors; Toxic effect; Translating; Xenograft Model; anticancer activity; cancer cell; cancer therapy; cell type; chemotherapeutic agent; chemotherapy; clinical care; common symptom; cytokine; embryonic stem cell; exosome; feasibility testing; improved; in vivo; knock-down; muscle strength; neoplastic cell; novel; p38 Mitogen Activated Protein Kinase; side effect; stem cell exosomes; systemic inflammatory response; tumor; tumor growth

Project Summary: Doxorubicin (DOX) is one of the most effective chemotherapeutic agents used for cancer treatment, but it also causes systemic inflammation and serious multi-organ side effects and muscle dysfunction in many patients. The loss of muscle strength in combination with constant fatigue is a burden on cancer patients undergoing chemotherapy. However, it is unknown whether inflammation induced cell death, known as pyroptosis (a distinct form of cell death, compared to apoptosis, and necrosis) occurs in DOX induced muscle toxicity and dysfunction (DIMT). Our preliminary data shows that treatment with DOX induces pyroptosis in Soleus muscle (SM), Sol 8 cells which is inhibited by exosomes derived from embryonic stem cells (ES-exos). We also observed increase in pro-inflammatory M1 macrophages, toll-like receptor 4 (TLR4), formation of NLRP3 inflammasome and pyroptotic specific caspase 1, IL-1β, and IL-18 post-DIMT in SM following DOX treatment. Mice treated with ES-Exos post-DMIT decreased M1 macrophages, TLR4, NLRP3 inflammasome and SM pyroptosis. Moreover, phenoswitching of M1 macrophages occurred which significantly increased anti- inflammatory M2 macrophages and reduced pyroptosis as well as adverse SM remodeling. miRNA array data shows increase in miR200c in ES-Exos. Furthermore, our preliminary data shows significantly decrease in phosphorylated p38 and JNK in DIMT following ES-Exos treatment, suggesting involvement of p38/JNK pathway. Therefore, based on this compelling data, we hypothesize that ES-Exos ameliorate DOX-induced pyroptosis and associated adverse muscle remodeling by increasing anti-inflammatory M2 macrophages thereby improving muscle function post-DIMT. We propose to test this hypothesis through the following 3 specific aims. Aim 1: To determine if increased level of M1 macrophages induce TLR4 and activates inflammasome (NLRP3) mediated pyroptosis during short and long-term post-DIMT. Aim 2 To demonstrate whether treatment with ES-Exos cause phenoswitching of M1 into M2 macrophages results in amelioration of pyroptosis and adverse SM remodeling. We will also determine the mechanisms of miR200c mediated p38 and JNK cell signaling in pyroptosis following ES-Exos treatment in DIMT. Additionally, we will examine the presence of broader range of miRNA specific for cell signaling, anti-pyroptotic, and anti-fibrotic pathways following ES-Exos treatment. Aim 3: To determine whether ES-Exos exert effects on anti-tumor efficacy of DOX in cancer while improving SM function. The proposed studies are hypothesis-driven, mechanistic, and should help define the impact of ES-Exos on M2 macrophage phenoswitching, pro- and anti-inflammatory cytokines, pyroptosis and apoptosis in the progression of DIMT. Also, we expect that ES-Exos would have no interference with the DOX-induced killing of cancer cells and inhibition of tumor growth. We believe that results obtained from these studies can easily be translated to the clinic as the cell free preparation of exosomes will not form teratomas, which is common occurrence with the use of human and mouse ES cells.

项目摘要： 阿霉素（DOX）是用于癌症治疗的最有效的化学治疗剂之一，但也是 导致许多患者的全身性炎症和严重的多器官副作用和肌肉功能障碍。 肌肉力量与持续疲劳结合的肌肉力量是对正在进行的癌症患者的伯嫩 化学疗法。然而，尚不清楚炎症是否诱导细胞死亡，称为凋亡（a 与细胞凋亡相比，细胞死亡的不同形式发生在DOX诱导的肌肉毒性和 功能障碍（DIMT）。我们的初步数据表明，用DOX治疗可诱导比目鱼的凋亡 肌肉（SM），SOL 8细胞，被源自胚胎干细胞（ES-EXOS）的外泌体抑制。 还观察到促炎性M1巨噬细胞的增加，Toll样受体4（TLR4），NLRP3的形成 DOX治疗后，SM的炎性体和凋亡特异性caspase 1，IL-1β和IL-18。 用ES-EXOS治疗的小鼠DMIT减少了M1巨噬细胞，TLR4，NLRP3炎症体和SM 凋亡。此外，发生了M1巨噬细胞的现象开关，这显着增加了抗 - 炎症性M2巨噬细胞和降低的投入性和广告SM重塑。 miRNA阵列数据 显示ES-EXOS中MIR200C的增加。此外，我们的初步数据显示 ES-EXOS处理后，DIMT的磷酸化p38和JNK，表明p38/jnk参与 路径。因此，基于此引人入胜的数据，我们假设ES-ES-exos可以改善DOX诱导 通过增加抗炎的M2巨噬细胞，凋亡和相关的不良肌肉重塑 从而改善dimt后的肌肉功能。我们建议通过以下3来检验这一假设3 具体目标。目标1：确定M1巨噬细胞的水平是否增加诱导TLR4并激活 炎症体（NLRP3）在DIMT短期和长期介导的凋亡。瞄准2证明 ES-EXO的处理是否导致M1进行M2巨噬细胞的现象，从而改善 凋亡和不良SM重塑。我们还将确定miR200C介导的p38和 DIMT的ES-EXOS治疗后，JNK细胞信号传导。此外，我们将研究 存在针对细胞信号，抗凋亡和抗纤维化途径的更广泛的miRNA范围 经过ES-EXOS处理。目标3：确定ES-EXO是否对抗肿瘤有效性产生影响 在改善SM功能的同时，癌症中的DOX。拟议的研究是由假设驱动的，机械和 应该有助于定义ES-EXO对M2巨噬细胞运转，促和抗炎的影响 DIMT进展中的细胞因子，凋亡和凋亡。另外，我们希望ES-Exos没有 干扰DOX诱导的癌细胞杀死和抑制肿瘤生长。我们相信 从这些研究中获得的结果可以轻松地转化为诊所，因为 外泌体不会形成畸胎瘤，这是使用人和小鼠ES细胞的常见发生。