Cancer Biology Research Test-Bed Unit 1: Effects of cell-intrinsic and cell-extrinsic signaling and mechanics on metastasis patterns of pediatric sarcomas

癌症生物学研究试验台单元 1：细胞内在和细胞外在信号传导和机制对儿科肉瘤转移模式的影响

• 批准号: 10684864
• 负责人: JAMES F AMATRUDA
• 金额: $ 39.11万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-24 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10684864
• 关键词: Adolescent and Young Adult; Affect; Applied Genetics; Beds; Behavior; Biological Assay; Biosensor; Bone Tissue; Buffers; Cancer Biology; Caveolins; Cell Survival; Cells; Cellular Morphology; Child; Clinical Trials; Collaborations; Complement; Data; Disease; Disseminated Malignant Neoplasm; Embryo; Enabling Factors; Engraftment; Environment; Epigenetic Process; Event; Ewings sarcoma; Failure; Fishes; Genetic; Genetic Models; Genome; Genomics; Goals; Heterogeneity; Human; Image; Imaging technology; In Situ; Intrinsic factor; Link; MAP Kinase Gene; Malignant Bone Neoplasm; Mechanics; Membrane; Metabolic; Metastatic/Recurrent; Microscopy; Morphology; Multimodal Imaging; Mus; Neoplasm Metastasis; Optics; Organ; Organism; Outcome; Oxygen; Pathway interactions; Patients; Pattern; Phenotype; Population; Primary Neoplasm; Property; Recurrence; Research; Resolution; Role; Signal Pathway; Signal Transduction; Site; Survival Rate; System; Testing; Tissues; Treatment-related toxicity; Visit; Visualization; WNT Signaling Pathway; Whole Organism; Xenograft Model; Xenograft procedure; Zebrafish; adverse outcome; cancer cell; caveolin 1; cell behavior; chemotherapy; childhood sarcoma; driver mutation; functional adaptation; genetic manipulation; high resolution imaging; imaging capabilities; in vivo; insight; mouse model; multimodality; neoplastic cell; novel strategies; optogenetics; prevent; quantitative imaging; sarcoma; soft tissue; tool; tumor; tumor microenvironment; tumor xenograft

Ewing sarcoma, a malignant tumor of bone and soft tissue affecting children, adolescents, and young adults. For the one-third of Ewing sarcoma patients who develop metastasis, the long-term survival rate remains less than 30%. Decades of clinical trials with ever-increasing intensity of chemotherapy have increased the toxicity of treatment but have not affected the poor outcome of metastatic disease. This failure to adequately treat metastases indicates that new approaches are needed to better understand the genesis of metastatic cells from the primary tumor and behavior of these cells in the in vivo microenvironment. Though there has been substantial progress in genomic profiling of tumors, these assays are unlikely to identify major determinants of metastatic behavior. This is because i) Ewing sarcomas typically have “quiet” genomes with few identifiable driver mutations; and ii) adverse outcomes may arise due to the functional adaptations of a small population of cells to the tumor microenvironment, driven by epigenetic, metabolic, morphologic or non-cell autonomous mechanisms. The broad goal of this project is thus to determine how extrinsic and intrinsic factors influence Ewing sarcoma cell fates at the metastatic site. A significant barrier to better understanding has been the lack of experimental systems that can probe heterogeneity of cell functional states, at whole-organism, single-cell and subcellular levels. Recent findings suggest that modulation of cell-mechanical features via the caveolin-1 and WNT signaling pathways may contribute to Ewing sarcoma metastasis, however the mechanisms of this adaptation are not known. As a system to visualize the heterogeneous functional properties of the metastatic cell population shed from a primary tumor, we leverage the zebrafish embryo as a host organism for human tumor xenografts. Ewing sarcoma cells readily engraft into zebrafish embryos and directly interact with the microenvironment of fully functional organs. The optical clarity of the fish allows us to perform multi-modal imaging to 1) identify host tissues associated with recurrent metastatic events; 2) define morphologic changes in cells undergoing metastatic adaptation in vivo; and 3) probe the activity of cancer cell signaling pathways in metastatic cells at subcellular resolution. Using powerful genetic tools and specific biosensors, we will exploit the quantitative imaging technology developed by TDU-1 to probe the role of caveolin-1 and WNT-dependent signaling in Ewing sarcoma metastasis. These studies will be complemented by parallel assays in mouse models and human Ewing sarcoma tumors, enabled by collaboration with TDU-2. Ultimately, these findings will inform strategies aimed at preventing or eliminating metastasis via targeting signaling mechanisms. We will determine effects of microenvironmental interactions on morphology and signaling of metastatic tumor cells; test the contribution of Caveolin-1 to metastatic cell adaptation to host environments; and probe the role of WNT signaling in Ewing sarcoma metastasis in genetic models. Ultimately these results will inform novel strategies to prevent or ameliorate metastasis in patients with Ewing sarcoma.

尤因肉瘤，一种影响儿童、青少年和年轻人的骨和软组织恶性肿瘤。 三分之一的尤文肉瘤患者发生转移，长期生存率仍然低于 数十年的临床试验和不断增加的化疗强度增加了毒性。 治疗但没有影响转移性疾病的不良结果。 转移表明需要新的方法来更好地了解转移细胞的起源 原发肿瘤和这些细胞在体内微环境中的行为已经存在很大的差异。 尽管肿瘤基因组分析取得了进展，但这些测定不太可能确定转移的主要决定因素 这是因为 i) 尤文肉瘤通常具有“安静”的基因组，几乎没有可识别的驱动因素。 突变；和 ii) 由于一小部分细胞的功能适应，可能会出现不良结果。 由表观遗传、代谢、形态或非细胞自主机制驱动的肿瘤微环境。 因此，该项目的总体目标是确定外在和内在因素如何影响尤文肉瘤 缺乏实验是更好地理解转移部位细胞命运的一个重大障碍。 可以探测整个生物体、单细胞和亚细胞细胞功能状态异质性的系统 最近的研究结果表明，通过 Caveolin-1 和 WNT 信号传导调节细胞机械特征。 途径可能有助于尤文肉瘤转移，但这种适应机制并不 作为一种可视化转移细胞群异质功能特性的系统。 从原发性肿瘤中，我们利用斑马鱼胚胎作为人类肿瘤异种移植物的宿主生物。 肉瘤细胞很容易植入斑马鱼胚胎并直接与斑马鱼的微环境相互作用 鱼的光学清晰度使我们能够进行多模态成像，以 1) 识别宿主。 与复发性转移事件相关的组织；2) 定义经历的细胞的形态变化 体内转移适应；3) 探测转移细胞中癌细胞信号通路的活性 使用强大的遗传工具和特定的生物传感器，我们将利用定量方法。 TDU-1 开发的成像技术用于探究 Caveolin-1 和 WNT 依赖性信号传导在尤文中的作用 这些研究将通过小鼠模型和人类尤因的平行测定来补充。 通过与 TDU-2 合作，这些发现最终将为针对肉瘤肿瘤的策略提供信息。 我们将确定通过靶向信号机制预防或消除转移的效果。 微环境相互作用对转移性肿瘤细胞形态和信号传导的影响； Caveolin-1 促进转移细胞适应宿主环境；并探讨 WNT 信号传导在 Ewing 中的作用； 最终，这些结果将为预防或预防肉瘤转移提供新的策略。 改善尤文肉瘤患者的转移。