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）：预计今年胰腺癌将在美国导致超过 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，以及缺乏治愈的治疗选择。传统癌症治疗的一个并发症是它们很容易错过一小部分细胞，称为干细胞，这些细胞已被证明与肿瘤的增殖能力有关。异常的蛋白质糖基化与癌症的发生和进展有关。特别是，具有唾液酸和硫酸基团的酸性聚糖经常被改变。有趣的是，最近发现的胰腺癌干细胞的细胞表面标志物是糖蛋白。然而，由于与不稳定、异质和支链聚糖的结构测定相关的巨大分析挑战，产生新型诊断和治疗（包括癌症疫苗）所需的详细的癌症相关聚糖结构很少。质谱 (MS) 可以提供灵敏且准确的聚糖分析。然而，酸性分子质谱的主要挑战是电离效率低。第二个挑战是确定糖分枝和特异性连接。此外，唾液酸和硫酸根基团极其不稳定，进一步损害电离并使硫酸根定位变得困难。该应用重点开发新的 MS 方法，用于识别和结构表征胰腺癌干细胞独特表达的聚糖（潜在生物标志物）。具体来说，氧化锆和二氧化钛表面化学将用于从复杂混合物中富集唾液酸化和硫酸化聚糖，从而大大改善纳米级正相液相色谱傅里叶变换离子回旋共振（FTICR）MS对它们的检测。为了确定这些聚糖的结构，我们将分别利用金属辅助电子捕获解离、电子脱离解离和真空紫外光解离来增加糖跨环裂解，从而提供连锁信息，从而可以确定支链结构，并确定硫酸盐位置。 公众健康相关性：胰腺癌是美国的主要死亡原因，五年生存率<5%。这项研究的重点是开发新方法，以改进胰腺癌干细胞表面碳水化合物（糖分子）的检测和结构测定，胰腺癌干细胞是肿瘤内负责其生长和增殖能力的细胞子集。众所周知，碳水化合物在癌症中会发生改变，因此成为癌症疫苗开发的有希望的目标。