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: 9755370
• 负责人: Muller Fabbri
• 金额: $ 31.01万
• 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 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9755370
• 关键词: Address; Affect; Back; Binding; Biological; Biology; Blood; Cancer Patient; Categories; Cells; 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; Neuroblastoma; Outcome; Patients; Pediatric Neoplasm; Pharmaceutical Preparations; Population; Primary Neoplasm; Proteins; Publishing; RHOB gene; 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; outcome forecast; 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.

项目摘要 儿童肿瘤是一个非常沉重的社会和经济负担， 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）是特别相关的，因为它是儿童以外最常见的实体癌。 它仍然杀死了大约40%的被诊断为最具攻击性形式的患者。因此有 显然需要改善对这种致命疾病的治疗。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细胞提供了阳性支持， 增加他们对化疗的抵抗力。调解这种支持和沟通的机制 这些细胞类型之间的媒介物知之甚少。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，并能与Toll样受体8（TLR 8）结合，触发TLR 8活化， 3）TLR 8刺激导致巨噬细胞分泌含有特异性miR的外泌体; 4） NB细胞摄取TAM衍生的外泌体miR，其使NB细胞中的BRG 1、RHOB和FBXW 7沉默。这些 蛋白质是MYC（又名c-MYC）和MYCN在NB中表达的众所周知的抑制剂; 5）MYC的水平 和MYCN在与人单核细胞共培养的NB细胞中以及在与人单核细胞共培养的原代NB中显著更高。 较高的TAM浸润。NB是一种非常MYC驱动的肿瘤。已知MYCN增加多药耐药性 在NB。我们提供的证据表明，MYC也促进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， 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），其诱导细胞增殖。 调节NB细胞中的MYC和MYCN，增加其对化疗的抗性。该提案有三个 具体目标。在第一部分中，我们将研究外泌体miRs诱导抗肿瘤的机制。 NB治疗然后我们将评估靶向TAM衍生的可溶性因子的治疗潜力 （外泌体miR和细胞因子）来克服NB抗性。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中TAM浸润程度有关，传统临床 成像技术和临床结果。我召集了一队经验丰富的科学家- 在NB以及miR和癌症治疗领域建立了专业知识。这项研究将导致 确定新的分子靶点，开发克服NB耐药性的新策略 (and可能是其他类型的MYC驱动的癌症），以及新的循环生物标志物的可能定义， 鉴定适合于抗TAM治疗的NB患者的子集。