Glycocalyx and NCOR2-Notch-Mediated Stemness in Glioblastoma Aggressiveness

糖萼和 NCOR2-Notch 介导的胶质母细胞瘤侵袭性干性

• 批准号: 8906462
• 负责人: Yekaterina Andreevna Miroshnikova
• 金额: $ 1.14万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8906462
• 关键词: Address; Adult Glioma; Aggressive behavior; Apoptosis; Automobile Driving; Behavior; Biochemical; Biocompatible Materials; Biological Assay; Bioluminescence; Biomechanics; Biophysical Process; Biophysics; Brain; Brain Stem; Breast; Cancer Biology; Carbohydrates; Caring; Cell Survival; Cell surface; Cells; Cessation of life; Clinical; Computer Simulation; Confocal Microscopy; Consensus; Critical Pathways; Deposition; Development; Diffuse; Dominant-Negative Mutation; Effectiveness; Engineering; Epithelial Cells; Focal Adhesions; Fostering; Generations; Genes; Genetic; Genetic Engineering; Genetic Transcription; Glioblastoma; Glycocalyx; Glycoproteins; Goals; HDAC3 gene; Health; Human; Hyaluronic Acid; Image; In Vitro; Infiltration; Integrins; Lasers; Lateral; Lead; Life; Link; Luciferases; Malignant Neoplasms; Mean Survival Times; Measures; Mechanics; Mediating; Messenger RNA; Mission; Modality; Modeling; Molecular; Monitor; Mus; Myosin ATPase; NCOR2 gene; Nature; Patients; Phenotype; Property; Proteins; Quality of life; Radio; Relapse; Relative (related person); Reporter; Research; Research Personnel; Resistance; Role; Scanning; Scientist; Signal Pathway; Signal Transduction; Stem cells; Survival Rate; Techniques; Testing; Therapeutic; Tissues; Traction; Tumor Cell Invasion; Up-Regulation; Work; Xenograft procedure; base; biophysical properties; brain tissue; caspase-3; cell behavior; cell growth; chemotherapy; cohort; design; hyaluronan synthase 1; improved; in vivo; loss of function; luminescence; mouse model; mutant; neoplastic cell; notch protein; novel; overexpression; prevent; public health relevance; rapid growth; receptor; research study; response; second harmonic; secretase; small hairpin RNA; stem; stemness; temozolomide; therapy resistant; tumor; tumor progression

DESCRIPTION (provided by applicant): Patients with high grade glioblastoma multiforme (GBM) have a mean survival time of 6-12 months due to the relative ineffectiveness of radio and chemotherapy and diffuse invasion into healthy brain tissue. Although the exact "apex cell" of origin for any GBM subtype remains uncertain, there is reasonable consensus that the most aggressive GBMs arise from stem cells, or at the very least from pluripotent cells. Overwhelming majority of GBM research to date has focused on GBM cell's unique genetic and biochemical signaling components with little or no focus on the biophysical features. Work proposed here aims to characterize the cell-intrinsic biophysical features of GBM cells that render them aggressive. Successful completion of this interdisciplinary work has the potential to offer a new paradigm with which to clarify the basis for enhanced survival, invasiveness, and treatment resistance of GBMs. Objectives: The long term goal of proposed work is to reveal more effective GBM treatment modalities, extend patient survival, and improve the quality of life for GBM patients by integrating physical scientist's concepts with the basic and clinical cancer biology research in order to explore the role of cell-intrinsic force in GBM aggression. Specifically, since aggressive nature of GBMs has been attributed to their stem-like properties, the proposal tests the idea that stemness and death-resistant behavior is mediated though a unique cell-intrinsic mechano-phenotype due to altered glycocalyx, the cell-associated carbohydrate layer, and interrogates the molecular mechanism responsible for its upregulation, which is hypothesized to act through NCoR2 and Notch. Overview of Approach: Proposed work tests a functional link between Notch and NCoR2 signaling and elucidates whether their combined action synergistically drives deposition of high levels glycoproteins at the cell surface which can in turn drive integrin and notch activation by biophysical means to modify survival, invasion, and treatment resistance. Techniques such as traction force, scanning angle interference, TIRF, second harmonic generation, spinning disc, and laser confocal microscopies will be used to characterize the biophysical properties of WT human GBM cells as well those with modified NCoR2 or Notch. To interrogate the mechanism sustaining these stem-like properties in vitro, genetic engineering of GBM cells and biomaterial engineering strategies will be used. Orthotopic xenograft mouse models of human GBMs will be employed to test the effectiveness of targeting NCoR2-Notch-glycocalyx circuitry in vivo.

描述（申请人提供）：由于放疗和化疗的相对无效性以及对健康脑组织的弥漫性侵袭，高级别多形性胶质母细胞瘤（GBM）患者的平均生存时间为 6-12 个月。尽管任何 GBM 亚型的确切“顶端细胞”起源仍不确定，但有合理的共识认为，最具侵袭性的 GBM 来自干细胞，或者至少来自多能细胞。迄今为止，绝大多数 GBM 研究都集中在 GBM 细胞独特的遗传和生化信号成分上，很少或根本不关注生物物理特征。这里提出的工作旨在表征 GBM 细胞的细胞固有生物物理特征，这些特征使它们具有攻击性。这项跨学科工作的成功完成有可能提供一个新的范式，以阐明提高 GBM 的生存率、侵袭性和治疗耐药性的基础。目的：拟议工作的长期目标是通过将物理科学家的概念与基础和临床癌症生物学研究相结合，揭示更有效的 GBM 治疗方式，延长患者生存期，提高 GBM 患者的生活质量，以探索其作用GBM 攻击中的细胞内在力。具体来说，由于 GBM 的攻击性归因于它们的干细胞样特性，因此该提案测试了这样的观点，即干细胞特性和抗死亡行为是通过由于改变的糖萼（细胞相关碳水化合物）而产生的独特的细胞固有机械表型介导的。层，并询问导致其上调的分子机制，假设该机制通过 NCoR2 和 Notch 发挥作用。方法概述：拟议的工作测试了 Notch 和 NCoR2 信号传导之间的功能联系，并阐明它们的联合作用是否协同驱动高水平糖蛋白在细胞表面的沉积，进而通过生物物理手段驱动整合素和 notch 激活，从而改变生存、侵袭、和治疗抵抗力。牵引力、扫描角干涉、TIRF、二次谐波发生、旋转盘和激光共焦显微镜等技术将用于表征 WT 人类 GBM 细胞以及具有修饰的 NCoR2 或 Notch 的细胞的生物物理特性。为了探究在体外维持这些干细胞样特性的机制，将使用 GBM 细胞的基因工程和生物材料工程策略。人类 GBM 的原位异种移植小鼠模型将用于测试体内靶向 NCoR2-Notch-糖萼电路的有效性。