NATURAL KILLER CELL BASED IMMUNOTHERAPY

我们的长期目标是开发用于高危神经母细胞瘤的免疫疗法， 自然杀伤（NK）细胞与抗肿瘤单克隆抗体（例如，anfi-GD 2）根除 原发性和转移性疾病。我们假设：1）NK抗体的抗肿瘤功效 依赖性细胞毒性（ADCC）取决于细胞因子的数量、持久性和抗肿瘤功能。 肿瘤微环境中的NK细胞。2)神经母细胞瘤细胞和单核吞噬细胞 合作创造一个微环境，其中包括IL-6和TGFB 1，这是促肿瘤和 免疫抑制因此，治疗策略必须使ADCC和细胞因子的NK活化最大化。 分泌（例如，Ifni）和最小化微环境抑制（例如，IL-6和TGFB 1）。我们的具体 目的如下：1）鉴定肿瘤细胞和单核吞噬细胞之间的相互作用， 肿瘤生长和/或抑制抗肿瘤NK功能。2)开发组合疗法， 通过最大化NK细胞的抗肿瘤活性，同时消除NK细胞的促肿瘤活性， 单核吞噬细胞研究设计和方法。在目标1中。我们将使用现有的微阵列 数据库以获得对来自患者的肿瘤的炎症相关基因表达的进一步了解（例如，白介素- 6/STAT 3和TGFB途径），然后用蛋白质阵列分析验证发现。接下来，我们将使用 了解肿瘤细胞和单核细胞之间的相互作用，增加肿瘤生长的体外模型 抑制NK细胞对ADCC和Ifni分泌的活化。最后，我们将使用体内模型， NOD/SCID小鼠中的局部和播散性疾病，包括肿瘤细胞的生物发光成像 单核细胞和NK细胞，以进一步了解单核细胞如何促进肿瘤生长， 它们引起对NK ADCC的抗性。在目标2中，我们将开发组合疗法， NK ADCC，同时否定肿瘤细胞的促肿瘤和免疫抑制作用， 单核吞噬细胞在这里，我们将使用体外模型来开发药物策略，例如 免疫调节剂来那度胺用于最大化NK ADCC诱导和细胞因子分泌， 调节单核细胞的促肿瘤功能。这一目标将包括产生微阵列数据，用于 连通性映射旨在识别用于免疫调节的新试剂。最有前途的战略 然后将与我们的体内NOD/SCID模型进行测试，其中包括成像，最小和建立 传播性和局部性疾病。这些实验将为临床试验开发提供基础。

Our long-term goal is to develop immunotherapy for high-risk neuroblastoma that realizes the full potential of natural killer (NK) cells combined with antitumor monoclonal antibodies (e.g., anfi-GD2) to eradicate primary and metastatic disease. We hypothesize the following: 1) Anti-tumor efficacy of NK antibody dependent cellular cytotoxicity (ADCC) depends upon the quantity, persistence, and anti-tumor function of NK cells in tumor microenvironments. 2) Neuroblastoma cells and mononuclear phagocytes reciprocally cooperate to create a microenvironment milieu, which includes IL-6 and TGFB1, that is pro-tumor and immunosuppressive. Thus, therapeutic strategies must both maximize NK activation for ADCC and cytokine secretion (e.g., Ifni) and minimize microenvironment suppression (e.g., IL-6 and TGFB1). Our Specific Aims are as follows: 1) Identify interactions between tumor cells and mononuclear phagocytes that promote tumor growth and/or suppress anti-tumor NK functions. 2) Develop combinatorial therapy for established and minimal disease by maximizing anti-tumor activities of NK cells while abrogating pro-tumor activities of mononuclear phagocytes. Research Design and Methods. In Aim 1. we shall use our existing microarray database to gain further insight into inflammation-related gene expression of tumors from patients (e.g., IL- 6/STAT3 and TGFB pathways) and then validate findings with protein array analyses. Next, we shall use in vitro models to understand interactions between tumor cells and monocytes that increase growth of tumor cells and suppress activation of NK cells for ADCC and Ifni secretion. Finally we shall use In vivo models of local and disseminated disease in NOD/SCID mice that include bioluminescent imaging of tumor cells monocytes, and NK cells to further understand how monocytes promote tumor growth and to determine if they cause resistance to NK ADCC. In Aim 2, we shall develop combinatorial therapy that maximizes antitumor NK ADCC while negating pro-tumor and immunosuppressive contributions from tumor cells and mononuclear phagocytes. Here, we shall use In vitro models to develop strategies with drugs such as the immune modulator lenalidomide for maximizing induction of NK ADCC and cytokine secretion while modulating the pro-tumor functions of monocytes. This aim will include generating microarray data for use in connectivity mapping aimed at identifying new agents for immune modulation. The most promising strategies then will be tested with our in vivo NOD/SCID models, which include imaging, of minimal and established disseminated and local disease. These experiments will provide the basis for clinical trial development.