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.

项目概要： 阿霉素 (DOX) 是用于癌症治疗的最有效的化疗药物之一，但它也 导致许多患者出现全身炎症、严重的多器官副作用和肌肉功能障碍。 肌肉力量的丧失加上持续的疲劳对癌症患者来说是一种负担 化疗。然而，尚不清楚炎症是否会诱导细胞死亡，即细胞焦亡（一种 与细胞凋亡和坏死相比，不同形式的细胞死亡发生在 DOX 诱导的肌肉毒性和 功能障碍（DIMT）。我们的初步数据表明，DOX 治疗会导致比目鱼肌焦亡 肌肉 (SM)、Sol 8 细胞，受胚胎干细胞 (ES-exos) 来源的外泌体抑制。我们 还观察到促炎性 M1 巨噬细胞、Toll 样受体 4 (TLR4) 以及 NLRP3 形成的增加 DOX 治疗后 SM 中 DIMT 后的炎症小体和焦亡特异性 caspase 1、IL-1β 和 IL-18。 DMIT 后接受 ES-Exos 治疗的小鼠 M1 巨噬细胞、TLR4、NLRP3 炎性体和 SM 减少 细胞焦亡。此外，M1巨噬细胞发生表型转换，显着增加抗- 炎症性 M2 巨噬细胞并减少焦亡以及不良 SM 重塑。 miRNA 阵列数据 显示 ES-Exos 中 miR200c 增加。此外，我们的初步数据显示， ES-Exos 处理后 DIMT 中 p38 和 JNK 磷酸化，表明 p38/JNK 参与其中 途径。因此，基于这些令人信服的数据，我们假设 ES-Exos 可以改善 DOX 诱导的 通过增加抗炎 M2 巨噬细胞导致焦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 个方面来检验这一假设 具体目标。目标 1：确定 M1 巨噬细胞水平增加是否会诱导 TLR4 并激活 炎症小体（NLRP3）在 DIMT 后短期和长期期间介导细胞焦亡。目标 2 展示 ES-Exos 治疗是否会导致 M1 巨噬细胞表型转换为 M2 巨噬细胞，从而改善 焦亡和不良 SM 重塑。我们还将确定 miR200c 介导的 p38 和 DIMT 中 ES-Exos 处理后焦亡中的 JNK 细胞信号传导。此外，我们将检查 存在更广泛的细胞信号转导、抗焦亡和抗纤维化途径特异的 miRNA ES-Exos 治疗后。目标3：确定ES-Exos是否对抗肿瘤功效产生影响 DOX在癌症中同时改善SM功能。拟议的研究是假设驱动的、机械的和 应该有助于确定 ES-Exos 对 M2 巨噬细胞表型转换、促炎和抗炎的影响 DIMT 进展中的细胞因子、细胞焦亡和细胞凋亡。此外，我们预计 ES-Exos 不会有 干扰 DOX 诱导的癌细胞杀伤和抑制肿瘤生长。我们相信 从这些研究中获得的结果可以很容易地转化为临床，因为无细胞制剂 外泌体不会形成畸胎瘤，而畸胎瘤在使用人类和小鼠 ES 细胞时很常见。