Role of microRNAs released by Tumor Associated Macrophages within Exosomes in the chemoresistance of Neuroblastoma

外泌体中肿瘤相关巨噬细胞释放的 microRNA 在神经母细胞瘤化疗耐药中的作用

• 批准号: 10165657
• 负责人: Muller Fabbri
• 金额: $ 1.17万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-26 至 2021-06-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10165657
• 关键词: Address; Affect; Back; Binding; Biological; Biology; Blood; Cancer Patient; Categories; Cells; Chemoresistance; Chemotherapy-Oncologic Procedure; Child; Cisplatin; Clinical; Communication; Data; Development; Diagnosis; Diagnostic radiologic examination; Disease; Drug resistance; Economic Burden; Emotional; FBXW7 gene; Family; Friends; Genes; Genomics; Goals; Grant; Growth; Human; Imaging Techniques; In Vitro; Infiltration; Inflammatory Response; Isotretinoin; Lead; MYCN gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; MicroRNAs; Molecular Target; Multi-Drug Resistance; Neoplasm Metastasis; Neuroblastoma; Outcome; Patients; Pediatric Neoplasm; Population; Primary Neoplasm; Prognosis; Proteins; Publishing; RHOB gene; Relapse; Research; Resistance; Role; SMARCA4 gene; Sampling; Scientist; Seasons; Solid; TLR8 gene; Therapeutic; Treatment Failure; Tumor-associated macrophages; Up-Regulation; c-myc Genes; cancer cell; cancer type; cell type; chemotherapy; circulating biomarkers; clinical imaging; cranium; cytokine; exosome; experimental study; high risk; high risk population; improved; in vivo; inhibitor/antagonist; loss of function; macrophage; monocyte; nanoparticle delivery; neuroblastoma cell; novel therapeutic intervention; patient response; promoter; resistance mechanism; social; standard of care; therapy resistant; tumor; tumor microenvironment; uptake

PROJECT SUMMARY Pediatric tumors represent a very heavy social and economic burden with profound emotional involvement not only for the directly affected children but also for their family and friends. Within pediatric tumors, neuroblastoma (NB) is of particular relevance, since it is the most common solid cancer in children outside of the skull and it still kills about 40% of patients diagnosed with the most aggressive forms. Therefore, there is a clear need to improve the treatment for this deadly disease. One of the strategies to achieve this goal is to overcome the emergence of resistance to chemotherapy by cancer cells. Published data show that Tumor-Associated Macrophages (TAMs) provide positive support to NB cells and increase their resistance to chemotherapy. The mechanisms that mediate this support and the communication vehicles between these cell types are poorly understood. My preliminary data have started to elucidate this, through the following findings: 1) NB cells secrete exosomes that contain microRNAs (miRs); 2) The miRs are taken up by surrounding macrophages and can bind to Toll-like receptor 8 (TLR8), triggering TLR8 activation in macrophages; 3) TLR8 stimulation causes macrophages to secrete exosomes that contain specific miRs; 4) the NB cells uptake TAM-derived exosomic miRs which silence BRG1, RHOB, and FBXW7 in NB cells. These proteins are well known inhibitors of MYC (aka as c-MYC) and MYCN expression in NB; 5) the levels of MYC and MYCN are significantly higher in NB cells co-cultured with human monocytes and in primary NBs with higher TAM infiltration. NB is a very MYC-driven tumor. It is known that MYCN increases multi-drug resistance in NB. We provide evidence that also MYC promotes NB resistance to chemotherapy both in vitro and in vivo. We also identified a panel of cytokines secreted by TAMs when co-cultured with NB cells, which could contribute to the up-regulation of MYC and MYCN in NB cells when co-cultured with human monocytes. Therefore, our hypothesis is that TAMs secrete soluble factors (exosomic miRs and cytokines) that induce up- regulation of MYC and MYCN in NB cells, increasing their resistance to chemotherapy. This proposal has three Specific Aims. In the first we will examine the mechanisms by which exosomic miRs induce resistance to therapy in NB. Then we will assess the therapeutic potential of targeting TAM-derived soluble factors (exosomic miRs and cytokines) to overcome NB resistance. Finally, we will correlate circulating TAM-derived exosomes in the blood of NB patients with the degree of TAM infiltration in the primary tumor, traditional clinical imaging techniques and clinical outcome. I have brought together a team of seasoned scientists with well- established expertise in NB and in the field of miRs and cancer therapeutics. This research will lead to the identification of new molecular targets, the development of a new strategy to overcome drug resistance in NB (and possibly other types of MYC-driven cancers), and the possible definition of new circulating biomarkers to identify subsets of NB patients suitable for an anti-TAM therapy.

项目概要 小儿肿瘤带来了非常沉重的社会和经济负担，并且具有深刻的情感影响 不仅是为了直接受影响的儿童，也是为了他们的家人和朋友。在儿科肿瘤中， 神经母细胞瘤（NB）尤其重要，因为它是除其他国家以外的儿童中最常见的实体癌症 它仍然杀死了大约 40% 被诊断患有最具侵袭性类型的患者。因此，有一个 显然需要改进这种致命疾病的治疗方法。实现这一目标的策略之一是 克服癌细胞对化疗产生的耐药性。 已发表的数据显示，肿瘤相关巨噬细胞（TAM）为 NB 细胞和 增加他们对化疗的抵抗力。调解这种支持和沟通的机制 人们对这些细胞类型之间的媒介知之甚少。我的初步数据已经开始阐明这一点， 通过以下发现：1）NB细胞分泌含有microRNA（miR）的外泌体； 2) miR 是 被周围的巨噬细胞吸收，并可以与 Toll 样受体 8 (TLR8) 结合，触发 TLR8 激活 巨噬细胞； 3）TLR8刺激导致巨噬细胞分泌含有特定miR的外泌体； 4） NB 细胞摄取 TAM 衍生的外泌体 miR，从而沉默 NB 细胞中的 BRG1、RHOB 和 FBXW7。这些 蛋白质是众所周知的 NB 中 MYC（又名 c-MYC）和 MYCN 表达的抑制剂； 5) MYC水平 和 MYCN 在与人单核细胞共培养的 NB 细胞以及与人单核细胞共培养的原代 NB 中显着较高 较高的 TAM 渗透。 NB 是一种高度由 MYC 驱动的肿瘤。众所周知，MYCN 会增加多重耐药性 在NB。我们提供的证据表明 MYC 还可以促进 NB 在体外和体内对化疗的耐药性。 我们还鉴定了 TAM 与 NB 细胞共培养时分泌的一组细胞因子，这可以 当与人单核细胞共培养时，有助于 NB 细胞中 MYC 和 MYCN 的上调。 因此，我们的假设是 TAM 分泌可溶性因子（外泌体 miR 和细胞因子），从而诱导上行- 调节 NB 细胞中的 MYC 和 MYCN，增加其对化疗的抵抗力。这个提案有三点 具体目标。首先，我们将研究外泌体 miR 诱导耐药性的机制。 NB 中的治疗。然后我们将评估针对 TAM 衍生可溶性因子的治疗潜力 （外泌体 miR 和细胞因子）克服 NB 耐药性。最后，我们将循环 TAM 衍生的 NB患者血液中的外泌体与原发肿瘤TAM浸润程度、传统临床 影像技术和临床结果。我召集了一支经验丰富的科学家团队 建立了 NB 以及 miR 和癌症治疗领域的专业知识。这项研究将导致 识别新的分子靶点，制定克服NB耐药性的新策略 （以及可能其他类型的 MYC 驱动的癌症），以及新的循环生物标志物的可能定义 确定适合抗 TAM 治疗的 NB 患者亚群。