Sphingosine-1-Phosphate Pathway Based Therapy for Neuroblastoma

1-磷酸鞘氨醇途径治疗神经母细胞瘤

• 批准号: 8527748
• 负责人: FERNANDO A FERRER
• 金额: $ 32.47万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-09 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8527748
• 关键词: 14-3-3 Proteins; Accounting; Angiogenic Factor; Apoptosis; Apoptotic; Award; Binding; Biological Availability; Biological Models; CCL2 gene; Caspase; Cell Death; Cell Line; Cell Survival; Cell physiology; Cells; Ceramides; Cessation of life; Chemotactic Factors; Child; Childhood Solid Neoplasm; Clinic; Collaborations; Combined Modality Therapy; Cytotoxic agent; Development; Disease; Drug usage; Equilibrium; Evaluation; Future; Generations; Genetic; Genetic Models; Goals; Grant; H218 Protein; Half-Life; Immunosuppressive Agents; Individual; Laboratories; Lipids; MYCN gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Modeling; Mus; Neuroblastoma; Pathway interactions; Process; Property; Receptor Inhibition; Regulation; Reliance; Signal Transduction; Solid Neoplasm; Specificity; Sphingosine; Sphingosine-1-Phosphate Receptor; Stromal Neoplasm; Testing; Therapeutic Effect; Topoisomerase Inhibitors; Transgenic Model; Transgenic Organisms; Translating; Treatment Efficacy; Vascular Endothelial Growth Factors; Weight; Work; Xenograft Model; Xenograft procedure; analog; angiogenesis; base; cancer therapy; cell growth; chemotherapy; computerized data processing; cytotoxicity; design; drug development; efficacy evaluation; extracellular; high risk; improved; in vivo; information gathering; inhibitor/antagonist; knock-down; macrophage; neoplastic cell; neuroblastoma cell; novel; novel strategies; prevent; research study; small hairpin RNA; sphingosine 1-phosphate; sphingosine kinase; therapeutic target; tumor; tumor microenvironment; tumor progression; tumorigenic

DESCRIPTION (provided by applicant): Neuroblastoma (NB) accounts for 15% of all childhood cancer deaths. Survival for children with high-risk disease remains below 40% and survival curves have plateaued despite treatment intensification strategies. As progress has stagnated, novel treatment approaches are clearly needed. Our laboratory has recently discovered that in NB two processes central to tumor progression are regulated by S1P signaling and can be targeted for therapeutic benefit. The first process concerns cell fate. Inside cells, the delicate balance between pro-survival (S1P) and apoptosis (sphingosine, ceramide) signaling is dictated by sphingosine kinases (SphK1 and SphK2). Neuroblastoma is unique among solid tumors we have evaluated in its disproportionate reliance on SphK2, which we strategically exploit, weighting the pendulum towards cell death. We have accomplished this by utilizing the sphingosine analogue FTY720, which is avidly and specifically phosphorylated by SphK2, thereby acting as a competitive inhibitor increasing pro-apoptotic sphingosine levels, while diminishing S1P levels. Secondly, we have found that extracellular S1P modulates tumor-stromal interactions. The S1P receptor, S1P2, contributes to the regulation of the tumor microenvironment in NB through expression of the potent macrophage chemoattractant MCP-1/CCL2, as well as, angiogenic factor VEGF. As we show, selective interference with this process disrupts establishment of a milieu favorable for tumor progression. Using this novel 2-pronged approach we demonstrate efficacy and synergy with current chemotherapy. Thus, our overarching hypothesis is that rational exploitation/inhibition of S1P signaling mechanisms will synergize with cytotoxic agents and result in enhanced treatment efficacy. This hypothesis is explored in 3 aims in this proposal. Aim 1: Evaluates the efficacy of FTY720 based NB tumor inhibition- In this aim we explore the hypothesis that FTY720 based interference with over-expressed SphK2 in NB cells results in enhanced apoptotic signaling through caspase independent pathways, favoring tumor cell death. These studies will evaluate FTY720 activity, alone and in combination with topoisomerse inhibitors across selected NB cell lines, xenografts and in the MYCN transgenic model. The mechanisms involved in FTY720 related cell death will also be defined as this information may aid in future drug development in aim 3. Aim 2: Utility of S1P2 interference/inhibition strategies for treating neuroblastoma-Through induction of MCP-1/CLL2 and VEGF, S1P2 regulates macrophage invasion and angiogenesis, respectively. We will test the therapeutic effect of S1P2 receptor inhibition alone (JTE-013) and in combination therapy against NB cell lines, xenografts and the MYCN transgenic model. To determine if S1P2 is critical to tumor development and progression we will use shRNA to knock down S1P2 in NB cell lines and xenograft models. Further, we will cross transgenic MYCN mice with S1p2-/- mice available in our lab, assessing the significance of S1P2 using a genetic model. Aim 3: Development and testing of FTY720 and JTE013 analogs with improved anti-cancer properties- The first generation drugs used in aims 1 and 2 have limitations and can be associated with off-target effects. In this final aim we will test analogs of FTY720, which are designed not to bind to the S1P receptors, to reduce immunosuppressive properties and other off target effects, yet are able to target intracellular SphK2 inhibition and thereby achieve tumor cytotoxicity. In the case of JTE-013, longer in-vivo half-life and enhanced inhibitory activity are focuses for 2nd generation compounds, several compounds designed with these endpoints are evaluated using our established models. The final aim will be performed in collaboration with Dr. Rolf Swenson of Arroyo Biosciences.

描述（由申请人提供）：神经母细胞瘤（NB）占所有儿童癌症死亡的15％。高风险疾病儿童的生存率仍低于40％，尽管采用了治疗加强策略，但生存曲线仍然存在。随着进步的停滞，显然需要新的治疗方法。我们的实验室最近发现，在NB中，肿瘤进展的两个过程受S1P信号的调节，可以针对治疗益处。第一个过程涉及细胞命运。里面 细胞，促生阳性（S1P）和凋亡（鞘氨醇，神经酰胺）信号之间的微妙平衡是由鞘氨醇激酶（SPHK1和SPHK2）决定的。神经母细胞瘤在我们对SPHK2的不成比例依赖中评估的实体瘤中是独一无二的，我们从策略上利用了它，将摆的摆缘加权细胞死亡。我们通过利用鞘氨醇的类似物FTY720来实现这一目标，该鞘氨酸类似物FTY720被SPHK2狂热，专门磷酸化，从而充当竞争性抑制剂，从而增加了促凋亡的鞘氨酸水平，同时降低了S1P水平。其次，我们发现细胞外S1P调节肿瘤 - 基质相互作用。 S1P受体S1P2通过表达有效的巨噬细胞化学吸引剂MCP-1/CCL2以及血管生成因子VEGF来促进NB中肿瘤微环境的调节。如我们所示，对这一过程的选择性干扰破坏了有利于肿瘤进展的环境。使用这种新颖的2种方法，我们证明了与当前化学疗法的功效和协同作用。因此，我们的总体假设是，S1P信号传导机制的理性剥削/抑制作用将与细胞毒性剂协同作用，并提高治疗功效。该提议的3个目标中探讨了这一假设。 AIM 1：评估基于FTY720的NB肿瘤抑制的功效 - 在此目的中，我们探讨了以下假设：基于FTY720在NB细胞中基于FTY720对过表达的SPHK2的干扰导致通过caspase独立途径增强了凋亡信号传导，从而增强了肿瘤细胞死亡。这些研究将单独评估FTY720活性，并与跨选定的NB细胞系，异种移植物和MYCN转基因模型中的拓扑异构体抑制剂结合使用。 FTY720相关细胞死亡中涉及的机制也将被定义为AIM 3中的未来药物开发。AIM2：S1P2干扰/抑制策略的实用性，用于治疗神经母细胞瘤的直接诱导MCP-1/Cll2和VEGF，S1P2，S1P2，S1P2，S1P2，S1P2调节了巨噬细胞的入侵和血管生成。我们将单独测试S1P2受体抑制作用（JTE-013）以及针对NB细胞系，异种移植物和MYCN转基因模型的治疗作用。为了确定S1P2对于肿瘤发展和进展至关重要，我们将使用SHRNA在NB细胞系和异种移植模型中击倒S1P2。此外，我们将使用实验室中的S1P2 - / - 小鼠穿越转基因MYCN小鼠，并使用遗传模型评估S1P2的重要性。 AIM 3：FTY720和JTE013的开发和测试具有改进的抗癌特性 - 目标1和2中使用的第一代药物具有局限性，并且可能与脱靶效应有关。在这个最终目标中，我们将测试FTY720的类似物，这些类似物的设计不绑定到 S1P受体，以降低免疫抑制特性和其他关闭靶标作用，但能够靶向细胞内SPHK2抑制，从而实现肿瘤细胞毒性。在JTE-013的情况下，较长的体内半衰期和增强的抑制活性是第二代化合物的重点，使用我们既定的模型评估了使用这些端点设计的几种化合物。最终目标将与Arroyo Biosciences的Rolf Swenson博士合作进行。