Targeting IL-6/IL-6R/STAT3 in EMDR in Neuroblastoma

耐药性是高危神经母细胞瘤患者未能治愈的重要因素，但几乎没有 已知微环境对这种癌症耐药的贡献。在协作中， 该项目的研究人员已经证明，骨髓间充质干细胞(BMMSC)和 肿瘤相关单核/巨噬细胞()与人类有共同的相互作用途径 导致耐药的神经母细胞瘤细胞。这个途径的中心是由这些受刺激的肿瘤细胞产生的IL-6 基质细胞及其下游靶信号转导通路。我们假设这种保护允许 肿瘤细胞存活并经历额外的基因和表观遗传改变，使它们 更具抵抗力。我们还假设，结合EMDR途径的抑制剂，如IL-6/IL-6- 6R/STAT3联合化疗或靶向治疗高危神经母细胞瘤患者将有所改善 多药耐药肿瘤细胞的治疗反应和减少疾病复发。Aim1将 使用药物敏感的神经母细胞瘤细胞系和骨髓间充质干细胞、成骨前细胞和 确定单核/巨噬细胞是否对细胞毒药物产生耐药性，以确定其作用机制 并测试通路抑制剂在恢复药物敏感性方面的作用。届时，实验将被 用取自4期神经母细胞瘤患者的含有肿瘤的骨髓样本进行验证。 骨髓来源的药物敏感神经母细胞瘤细胞株长期共培养的研究 我们将测试EMDR是否促进表观遗传变化，从而导致药物状态 已经独立于微环境的抗性。AIM 2将在#年评估EMDR机制 体内应用MYCN-NA高危神经母细胞瘤(NB-Tag)转基因模型 IL-6/IL-6R/STAT3介导的EMDR将被消融。目标3将检验阻断1L-6/IL-6的假设 神经母细胞瘤中负责EMDR的6R/STATS通路增强了对化疗的反应。 这些实验最初将测试免疫调节剂来那度胺和抗IL-6R的tociluzumab 抗体以及SI PRI和JAK2的小抑制物。项目1将在测试方面与项目2协作 SDF1a/CXCR4是否在神经母细胞瘤的EMDR中发挥作用，并与项目3一起确定 S1P/S1PR1在IL-6介导的EMDR及临床前检测S1PR1和JAK2抑制剂中的作用 神经母细胞瘤模型。我们期待我们的研究将为最初的治疗提供一个新的范例。 不仅是患有高危神经母细胞瘤的儿童，还有其他癌症患者。

Drug-resistance is an important factor of failure to cure patients with high-risk neuroblastoma, and little is known of the contribution of the microenvironment to drug resistance in this cancer. In collaboration, the investigators of this project have demonstrated that bone marrow mesenchymal stem cells (BMMSC) and tumor associated monocytes/macrophages (TAM) share a common pathway of interaction with human neuroblastoma cells that leads to drug resistance. Central to this pathway is IL-6 produced by these tumor cellstimulated stromal cells and its downstream target STATS. We hypothesize that such protection allows tumor cells to survive and to undergo additional genetic and epigenetic alterations that render them even more resistant. We also postulate that combining inhibitors of EMDR pathways like IL-6/IL- 6R/STAT3 with chemotherapy or targeted therapy in patients with high-risk neuroblastoma will improve therapeutic response and decrease disease recurrence with multidrug resistant tumor cells. Aim1 will use co-cultures of drug sensitive neuroblastoma cell lines and BMMSC, pre-osteoblasts, and monocytes/macrophages to determine if they cause resistance to cytotoxic drugs, to identify mechanisms of resistance, and to test the effect of pathway inhibitors in restoring drug sensitivity. Experiments will then be validated with tumor containing bone marrow samples obtained from patients with stage 4 neuroblastoma. Using long term co-cultures of drug sensitive neuroblastoma cell lines in the presence of bone marrow-derived stromal cells we shall test whether EMDR promotes epigenetic changes that will lead to a state of drug resistance that has become independent of the microenvironment. Aim 2 will evaluate EMDR mechanism in vivo using a transgenic model of MYCN-NA high-risk neuroblastoma (NB-Tag) in which key genes involved in lL-6/IL-6R/STAT3-mediated EMDR will be ablated. Aim 3 will test the hypothesis that blocking 1L-6/IL- 6R/STATS pathways responsible for EMDR in neuroblastoma enhances the response to chemotherapy. These experiments initially will test lenalidomide, an immune modulator, and tociluzumab, an anti-IL-6R antibody as well a small inhibitors of SI PRI and JAK2. Project 1 will collaborate with Project 2 in testing whether the SDF1a/CXCR4 plays a role in EMDR in neuroblastoma and with Project 3 in determining the contribution of S1P/S1PR1 to IL-6-mediated EMDR and in testing inhibitors of S1PR1 and JAK2 in preclinical models of neuroblastoma. We anticipate that our research will provide a new paradigm for the initial treatment not only of children with high-risk neuroblastoma but also of patients with other cancers.