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: 10374652
• 负责人: JAMES F AMATRUDA
• 金额: $ 33.77万
• 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/10374652
• 关键词: Adolescent and Young Adult; Affect; Beds; Behavior; Biological Assay; Biosensor; Buffers; Cancer Biology; Caveolins; Cell Survival; Cells; Cellular Morphology; Child; Clinical Trials; Collaborations; Complement; Connective and Soft Tissue Neoplasm; Data; Disease; Disseminated Malignant Neoplasm; Embryo; Enabling Factors; 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 to; 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; WNT Signaling Pathway; Whole Organism; Xenograft Model; Xenograft procedure; Zebrafish; adverse outcome; cancer cell; caveolin 1; cell behavior; cell stroma; 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; 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.

尤文肉瘤，一种影响儿童、青少年和年轻人的骨和软组织恶性肿瘤。为 三分之一的尤文肉瘤患者发生转移，长期生存率仍然低于 百分之三十数十年的临床试验，化疗强度不断增加， 治疗，但没有影响转移性疾病的不良结局。这种未能充分治疗 转移表明，需要新的方法来更好地了解转移细胞的起源， 原发性肿瘤和这些细胞在体内微环境中的行为。尽管有大量的 尽管在肿瘤基因组谱分析方面取得了进展，但这些测定不太可能确定转移性肿瘤的主要决定因素。 行为这是因为i）尤因肉瘤通常具有“安静”的基因组，几乎没有可识别的驱动因子 突变;和ii）由于小细胞群的功能适应性， 肿瘤微环境，由表观遗传、代谢、形态学或非细胞自主机制驱动。 因此，本项目的主要目标是确定外在和内在因素如何影响尤文肉瘤 转移部位的细胞命运。更好地理解的一个重要障碍是缺乏实验性的 系统可以探测细胞功能状态的异质性，在整个生物体，单细胞和亚细胞 程度.最近的研究结果表明，通过小窝蛋白-1和WNT信号转导调节细胞力学特征， 通路可能有助于尤文肉瘤转移，然而，这种适应的机制不是 知道的作为一个系统，以可视化的异质性功能特性的转移细胞群体脱落 从原发性肿瘤，我们利用斑马鱼胚胎作为人类肿瘤异种移植的宿主生物体。尤因 肉瘤细胞很容易移植到斑马鱼胚胎中，并直接与完全生长的微环境相互作用。 功能器官。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）定义经历转移的细胞中的形态学变化 体内转移适应;和3）探测转移性细胞中癌细胞信号传导途径的活性， 亚细胞分辨率使用强大的遗传工具和特定的生物传感器，我们将利用定量的 由TDU-1开发的成像技术，用于探测小窝蛋白-1和WNT依赖性信号传导在Ewing中的作用 肉瘤转移这些研究将通过在小鼠模型和人尤因中的平行测定来补充。 肉瘤肿瘤，通过与TDU-2合作实现。最终，这些发现将为旨在 通过靶向信号传导机制预防或消除转移。我们将确定 微环境相互作用对转移性肿瘤细胞的形态和信号传导的影响;测试 Caveolin-1对转移性细胞适应宿主环境的影响;并探讨WNT信号转导在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 改善尤文肉瘤患者的转移。