Novel Approaches for Structural Determination of Cancer Stem Cell Glycans

癌症干细胞聚糖结构测定的新方法

• 批准号: 7852331
• 负责人: KRISTINA HAKANSSON
• 金额: $ 23.62万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7852331
• 关键词: Binding; Biological Markers; Cancer Vaccine Related Development; Cancer Vaccines; Cancer cell line; Cancerous; Carbohydrates; Cause of Death; Cell Separation; Cell membrane; Cell surface; Cells; Cessation of life; Charge; Chemistry; Collection; Complex; Complex Mixtures; Complication; Coupled; Data; Detection; Development; Disease; Dissociation; Distant; Electrons; Electrospray Ionization; Flow Cytometry; Fourier transform ion cyclotron resonance; Future; Gases; Generations; Genomics; Glycoproteins; Human; Inorganic Sulfates; Ions; Knowledge; Link; Liquid Chromatography; Location; Lymphatic System; Malignant Neoplasms; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Membrane; Membrane Glycoproteins; Metals; Methods; Modeling; Molecular; Neoplasm Metastasis; Oligosaccharides; Organ; Pancreatic Adenocarcinoma; Phase; Phosphopeptides; Photons; Play; Polysaccharides; Population; Proliferating; Protein Glycosylation; Proteins; Proteomics; Reaction; Reporting; Research; Research Personnel; Resistance; Role; Screening for cancer; Sialic Acids; Sorting - Cell Movement; Stem cells; Structure; Surface; Surface Antigens; Survival Rate; Techniques; Therapeutic; Therapeutic Agents; Tissues; Titania; United States; Unspecified or Sulfate Ion Sulfates; Vaccines; Vacuum; Xenograft Model; Zirconium; base; cancer cell; cancer stem cell; cancer therapy; divalent metal; glycosylation; improved; ionization; killings; metal oxide; method development; mortality; nano; nano-electrospray; nanoscale; novel; novel diagnostics; novel strategies; outcome forecast; public health relevance; sugar; tumor; tumor progression; tumor xenograft; tumorigenic; ultraviolet; zirconium oxide

DESCRIPTION (provided by applicant): Pancreatic cancer is expected to cause >32,000 deaths in he United States this year. This high mortality is largely due to lack of reliable methods for early tumor detection, and lack of treatment options that produce a cure. One complication with traditional cancer treatments is that they can easily miss the small subset of cells, termed stem cells, which have been shown to be responsible for a tumor's ability to proliferate. Aberrant protein glycosylation is linked to the onset and progression of cancer. Particularly, acidic glycans with sialic acid and sulfate groups are often altered. Interestingly, recently identified cell surface markers of pancreatic cancer stem cells are glycoproteins. However, due to tremendous analytical challenges associated with structural determination of labile, heterogeneous and branched glycans, detailed cancer-associated glycan structures, which are required for generation of novel diagnostics and therapeutics, including cancer vaccines, are scarce. Mass spectrometry (MS) can provide sensitive and accurate glycan analysis. However, a major challenge in acidic molecule MS is low ionization efficiency. A second challenge is the determination of saccharide branching and specific linkage. Also, sialic acids and sulfate groups are extremely labile, further compromising ionization and rendering sulfate localization difficult. This application focuses on developing novel MS approaches for identifying and structurally characterizing glycans uniquely expressed by pancreatic cancer stem cells (potential biomarkers). Specifically, zirconia and titania surface chemistry will be utilized to enrich sialylated and sulfated glycans from complex mixtures, thereby greatly improving their detection by nano-scale normal phase liquid chromatography Fourier transform ion cyclotron resonance (FTICR) MS. For structural determination of these glycans, we will utilize metal-assisted electron capture dissociation, electron detachment dissociation, and vacuum ultraviolet photodissociation, respectively, to increase sugar cross-ring cleavage, which provides linkage information and therefore allows determination of branched structures, and to determine sulfate location. PUBLIC HEALTH RELEVANCE: Pancreatic cancer is a major death cause in the United States with a five-year survival rate of <5%. This research focuses on developing novel approaches for improved detection and structural determination of carbohydrates (sugar molecules) present on the surface of pancreatic cancer stem cells, that is, a subset of cells within a tumor that is responsible for its ability to grow and propagate. Carbohydrates are known to be altered in cancer and therefore constitute promising targets for cancer vaccine development.

描述（由申请人提供）：胰腺癌预计将在今年在美国造成32,000人死亡。这种高死亡率很大程度上是由于缺乏可靠的早期肿瘤检测方法，并且缺乏产生治愈方法的治疗选择。传统癌症治疗的一种并发症是，它们很容易错过一小部分细胞，称为干细胞，这些细胞已被证明是导致肿瘤增殖能力的原因。异常的蛋白质糖基化与癌症的发作和进展有关。特别是，含唾液酸和硫酸盐基团的酸性糖通常会改变。有趣的是，最近发现的胰腺癌干细胞的细胞表面标记是糖蛋白。然而，由于与不稳定，异质和分支聚糖的结构确定相关的巨大分析挑战，详细的癌症相关的聚糖结构是稀缺的新型诊断和治疗剂（包括癌症疫苗）所必需的。质谱（MS）可以提供灵敏，准确的聚糖分析。但是，酸性分子MS的主要挑战是低电离效率。第二个挑战是确定糖分支和特定连锁。同样，唾液酸和硫酸盐基团极不稳定，进一步损害了电离和使硫酸盐定位难以进行。该应用的重点是开发新的MS方法，用于识别和结构表征胰腺癌干细胞（潜在的生物标志物）独特表达的聚糖。具体而言，将利用氧化锆和钛表面化学来富集复杂混合物中的硫化和硫化的聚糖，从而通过纳米尺度正常相液相色谱傅立叶转化离子转化离子环体共振（FTICR）MS大大改善它们的检测。为了确定这些聚糖的结构性测定，我们将分别利用金属辅助电子捕获分离，电子脱离分离和真空紫外线光解异步来增加糖横环的裂解，从而提供链​​接信息，从而提供链​​接信息，从而允许确定分支结构，并确定分支结构，以及到达分支结构，以及用于分支结构的裂解。确定硫酸盐位置。 公共卫生相关性：胰腺癌是美国的主要死亡原因，五年生存率<5％。这项研究重点是开发新的方法，以改善存在于胰腺癌干细胞表面上的碳水化合物（糖分子）的结构测定，即肿瘤中细胞的子集，该子集负责其生长和繁殖能力。已知碳水化合物在癌症中发生改变，因此构成了癌症疫苗发育的有希望的靶标。