Proj 3 - Targeting the Pro-tumorigenic Microenvironment

总结/摘要 项目4的总体目标是通过以下方法发现和利用治疗抵抗的外在机制： 关注肿瘤相关巨噬细胞（TAM）和肿瘤相关成纤维细胞的贡献 肿瘤微环境（TME）中的TAF。我们的总体假设是，TAMs和TAFs合作 在创造一个有利的致瘤环境，最终导致治疗的出现， 抵抗和免疫逃逸。我们还假设，随着肿瘤的治疗，TME在其 组合物和功能变得越来越有利于治疗抗性。这个假设是基于 根据我们小组发表的初步数据， 在NB的炎症亚型中，在诊断时与复发的高风险和极差的 预后我们也有证据表明，TAM和TAF在暴露于肿瘤细胞时， 通过旁分泌产生促肿瘤细胞因子的增殖、存活和耐药性， 在肿瘤细胞中激活信号传导途径的趋化因子，如STAT 3和ERK。我们的项目有三个目标。 目标1，将审查技术援助机制和技术援助机制之间的合作机制，检验以下假设： MYCN扩增的肿瘤不产生TAM化学引诱物CCL-2/MCP-1，TAF是一个来源， 这个趋化因子我们还将研究细胞因子和趋化因子的贡献，在共培养产生的， TAMs、TAFs和NB细胞以及它们在NB细胞中激活的信号传导通路导致增加的细胞内信号传导。 增殖和生存。目的2，将检查继发于化疗的TME景观的变化， 同基因小鼠NB模型（与项目2），并验证通过Core获得的患者肿瘤样本中的数据 B。通过检测长期暴露于TAM/TAF的NB细胞中转录组的变化， 其潜在的表观遗传起源（与项目3），目标2还将确定脆弱性，以防止耐药性， 化疗或靶向治疗（项目1）。目标3，然后将这些发现转化为临床前 肿瘤模型我们将测试最有前途的药物靶向TAF，TAM，或 它们在肿瘤细胞中激活的途径与化疗或免疫疗法相结合（与项目5）， 在免疫缺陷小鼠以及鼠细胞系中使用人NB系和患者来源的异种移植物 在免疫活性小鼠中（项目2），然后将提出最有效的药物用于早期阶段 NANT（核心B）的临床试验。因此，项目4为实现这一总体目标作出了独特的贡献。 PPG通过其对TME和非自主机制的关注， 抵抗和免疫逃逸。

SUMMARY/ABSTRACT The overall objective of Project 4 is to discover and exploit extrinsic mechanisms of therapy resistance by focusing on the contribution of tumor-associated macrophages (TAMs) and tumor-associated fibroblasts (TAFs) in the tumor microenvironment (TME). Our overarching hypothesis is that TAMs and TAFs cooperate in creating a favorable tumorigenic environment that ultimately leads to the emergence of therapeutic resistance and immune escape in NB. We also postulate that as tumors are treated, the TME is altered in its composition and function to become increasingly favorable to therapeutic resistance. This hypothesis is based on published and preliminary data from our group demonstrating that TAMs and TAFs are abundantly present in an inflammatory subtype of NB at diagnosis associated with a high risk of recurrence and extremely poor prognosis. We also have evidence that TAMs and TAFs when exposed to tumor cells stimulate their proliferation, survival and drug-resistance via the paracrine production of pro-tumorigenic cytokines and chemokines that activate in tumor cells signaling pathways such as STAT3 and ERK. Our project has 3 aims. Aim 1, will examine mechanisms of cooperation between TAMs and TAFs, testing the hypothesis that in MYCN amplified tumors that do not produce the TAM chemoattractant CCL-2/MCP-1, TAFs are a source of this chemokine. We will also examine the contribution of cytokines and chemokines generated in co-culture of TAMs, TAFs and NB cells and the signaling pathways they activate in NB cells leading to increased proliferation and survival. Aim 2, will examine changes in the TME landscape secondary to chemotherapy in syngeneic murine NB models (with Project 2) and validate the data in patient tumor samples obtained via Core B. By examining changes in the transcriptome that occur in NB cells chronically exposed to TAM/TAF and their potential epigenetic origin (with Project 3), aim 2 will also identify vulnerabilities to prevent resistance to chemotherapy or targeted therapy (with Project 1). Aim 3, will then translate these discoveries in pre-clinical tumor models. We will test the therapeutic efficacy of the most promising agents targeting TAFs, TAMs, or pathways they activate in tumor cells in combination with chemotherapy or immunotherapy (with Project 5), using human NB lines and patient-derived xenotransplants in immunodeficient mice as well as murine cell lines in immunocompetent mice (with Project 2), The most effective agent(s) will then be proposed for early phase clinical trials to the NANT (Core B). Thus Project 4 brings a unique contribution to the overall objective of this PPG through its focus on the TME and on non-autonomous mechanisms leading towards therapeutic resistance and immune escape.