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

耐药是导致高危神经母细胞瘤患者无法治愈的重要因素， 已知微环境对这种癌症耐药性的贡献。在合作中， 该项目的研究人员已经证明，骨髓间充质干细胞（BMMSC）和 肿瘤相关单核细胞/巨噬细胞（TAM）与人类肿瘤细胞相互作用的共同途径 导致耐药性的神经母细胞瘤细胞。该途径的中心是由这些肿瘤细胞刺激产生的IL-6。 基质细胞及其下游靶点STATS。我们假设，这种保护允许 肿瘤细胞存活，并经历额外的遗传和表观遗传改变，使它们 甚至更有抵抗力我们还假设，联合使用EMDR通路的抑制剂，如IL-6/IL-10， 6 R/STAT 3在高危神经母细胞瘤患者中配合化疗或靶向治疗会有所改善 治疗反应和减少多药耐药肿瘤细胞的疾病复发。aim 1将 使用药物敏感性神经母细胞瘤细胞系和BMMSC、前成骨细胞的共培养物，和 单核细胞/巨噬细胞，以确定它们是否引起对细胞毒性药物的耐药性，以确定 抗性，并测试途径抑制剂在恢复药物敏感性中的作用。实验将在 用从患有4期神经母细胞瘤的患者获得的含有肿瘤的骨髓样品进行验证。 使用药物敏感性神经母细胞瘤细胞系在骨髓源性细胞因子存在下的长期共培养， 我们将测试EMDR是否会促进表观遗传变化，从而导致药物状态， 已经独立于微环境的抵抗力。目的2将评估EMDR机制， 体内使用MYCN-NA高危神经母细胞瘤（NB-Tag）的转基因模型，其中参与 IL-6/IL-6 R/STAT 3介导的EMDR将被消融。目的3将检验阻断1 L-6/IL-12表达的假设。 6 R/STATS通路负责神经母细胞瘤的EMDR增强对化疗的反应。 这些实验最初将测试免疫调节剂来那度胺和抗IL-6 R的tociluzumab 抗体以及SI PRI和JAK 2的小抑制剂。项目1将与项目2合作进行测试 SDF 1a/CXCR 4是否在神经母细胞瘤的EMDR中发挥作用，以及与项目3在确定 S1 P/S1 PR 1对IL-6介导的EMDR的贡献以及在临床前试验中测试S1 PR 1和JAK 2的抑制剂 神经母细胞瘤模型。我们期望我们的研究将为初始治疗提供新的范例 不仅是高危神经母细胞瘤患儿，也包括其他癌症患者。

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.