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，骨骼和软组织的恶性肿瘤，影响儿童，青少年和年轻人。为了 三分之一的尤因肉瘤患者发生转移，长期生存率仍然少于 30％。数十年的临床试验具有不断增加的化学疗法强度的毒性增加了 治疗但没有影响转移性疾病的不良结果。这种无法充分治疗 转移表明需要采用新的方法来更好地了解从 这些细胞在体内微环境中的主要肿瘤和行为。虽然有很大的 肿瘤基因组分析的进展，这些测定不可能鉴定转移性的主要决定剂 行为。这是因为i）尤因肉瘤通常具有几乎可识别驱动器的“安静”基因组 突变； ii）由于少数细胞群体对 肿瘤微环境由表观遗传学，代谢，形态学或非细胞自主机制驱动。 因此，该项目的广泛目标是确定外在因素和内在因素如何影响Ewing肉瘤 转移部位的细胞命运。更好地理解的一个重大障碍是缺乏实验 可以探测细胞功能状态的异质性的系统，在全虫，单细胞和亚细胞上 水平。最近的发现表明，通过小窝蛋白-1和Wnt信号调节细胞力学特征 途径可能有助于尤因肉瘤转移，但是这种适应的机制不是 已知。作为可视化转移细胞群的异质功能特性的系统 从原发性肿瘤中，我们利用斑马鱼的胚胎作为人类肿瘤Xenographictics的宿主生物。尤因 肉瘤细胞容易植入斑马鱼胚胎，并直接与完全相互作用 功能器官。鱼的光学清晰度使我们能够执行多模式成像至1）识别宿主 与复发转移事件相关的组织； 2）定义经历细胞的形态变化 体内转移适应； 3）探测转移细胞中癌细胞信号通路的活性 亚细胞分辨率。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 TDU-1开发的成像技术是为了探测Caveolin-1和Wnt依赖性信号在EWING中的作用 肉瘤转移。这些研究将通过鼠标模型和人类尤因中的并行测定完成 肉瘤肿瘤，通过与TDU-2合作实现。最终，这些发现将为针对的策略提供信息 通过靶向信号传导机制来防止或消除转移。我们将确定 关于转移性肿瘤细胞形态和信号传导的微环境相互作用；测试 Caveolin-1对宿主环境的转移细胞适应；并探测Wnt信号在EWING中的作用 遗传模型中的肉瘤转移。最终，这些结果将为预防或 Ewing肉瘤患者的改善转移。