Carbohydrate- Dependent Epithelial Cancer Metastasis

碳水化合物依赖性上皮癌转移

• 批准号: 7534124
• 负责人: Michiko Fukuda
• 金额: $ 22.9万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7534124
• 关键词: Anabolism; Annexin A1; Antibodies; Antigens; Apoptosis; Binding; Birth; Carbohydrates; Cell surface; Cells; Clinical; Clinical Research; Collaborations; Complex; Data; Embryo; Endothelial Cells; Epithelial; Epithelial Cells; Epitopes; Goals; Golgi Apparatus; Hematopoietic; Heparitin Sulfate; Heterogeneous Nuclear RNA; High Endothelial Venule; Isoenzymes; Kinetics; Knock-out; Knockout Mice; L-Selectin; Laboratories; Ligands; Lung; Malignant - descriptor; Malignant Neoplasms; Mannosidase; Mediating; Methods; Minor; Monoclonal Antibodies; Mus; Mutant Strains Mice; Neoplasm Metastasis; Oligosaccharides; P-Selectin; Peptide Phage Display Library; Peptide Receptor; Peptides; Personal Satisfaction; Polysaccharides; Process; Proteomics; RNA Splicing; Research; Role; Screening procedure; Selectins; Surface; Surface Plasmon Resonance; Tumor-Associated Carbohydrate Antigens; Wild Type Mouse; angiogenesis; base; cancer cell; in vivo; knockout gene; lung melanoma; mRNA Precursor; mannosyl-oligosaccharide 1,3-1,6-alpha-mannosidase; novel therapeutics; outcome forecast; tumor; tumor growth

It is well documented that malignant transformation of epithelial cells is associated with structural changes in cell surface carbohydrates. Many clinical studies have correlated those cancer-associated carbohydrate antigens with poor clinical prognosis, including metastasis. Carbohydrate-dependent cancer metastasis has been observed, but the mechanisms underlying this process are not yet defined. We previously identified IELLQAR (l-peptide) that mimics the epitope of anti-Lewis A antibody by screening peptide displaying phage libraries. When l-peptides were injected into wild type mice, they inhibited sLex-dependent melanoma lung colonization. However, lung colonization also occurs in mutant mice lacking both E- and P-selectins and l-peptide inhibits the colonization, excluding the involvement of these selectins in this process. These findings suggest that l-peptide interference with colonization does not require E- and P-selectins. Our preliminary data show that the l-peptide receptors are pre-mRNA splicing factor (Sfrs) and AnnexinAI (Anxal). The PI also studied the in vivo roles of the Golgi processing a-mannosidase II (Mil) and alphamannosidase llx (MX). Mil/MX double null mouse embryos showed no complex type A/-glycans, indicating that Mil and MX together are responsible for the complex type N-glycan synthesis. Mil/MX double nulls were lethal shortly after birth. Based on these findings, the specific aims are: (1) To determine A/-glycan-based L-selectin ligand activity in vivo using conditional Mil/MX double null mice. Endothelial and hematopoietic cell-specific Mil/MX double knockouts (Tie2-Cre:Mllftaxp/floxp/MX-/-) will be generated and analyzed for L-selectin ligand activity in high endothelial venules (HEVs), (2) to define carbohydrate-binding activity of Sfrs and Anxal, (3) to determine if targeted apoptosis occurs by Anxal binding. Anxal has been identified as a tumor-specific endothelial marker, and we identified IFpeptide, which binds preferentially to Anxal. Thus we will develop a method to target apoptosis to endothelial cells of tumor vasculature in the mouse using IF-peptide, and (4) to determine if Anxal expressed on the endothelial surface promotes angiogenesis and identify mechanisms underlying Anxal-dependent endothelial cell activation including binding to heparan sulfate. These studies will provide information for better understand of the mechanisms underlying carbohydrate-dependent cancer metastasis and help for developing new therapeutic strategies against epithelial cancer.

有充分证据表明，上皮细胞的恶性转化与结构性改变有关。 细胞表面碳水化合物的变化。许多临床研究已将这些与癌症相关的 碳水化合物抗原具有不良的临床预后，包括转移。碳水化合物依赖性 已经观察到癌症转移，但这一过程的机制尚未明确。 我们之前通过以下方法鉴定了模仿抗路易斯 A 抗体表位的 IELLQAR（l 肽）： 筛选肽展示噬菌体文库。当L-肽注射到野生型小鼠体内时，它们 抑制 sLex 依赖性黑色素瘤肺部定植。然而，肺部定植也发生在突变体中 缺乏E-和P-选择素以及L-肽的小鼠抑制定植，排除以下因素的参与 这些选择素在此过程中。这些发现表明，L-肽干扰定植确实 不需要E-和P-选择素。我们的初步数据表明，L-肽受体是前mRNA 剪接因子（Sfrs）和AnnexinAI（Anxal）。 PI 还研究了高尔基体加工 α-甘露糖苷酶 II (Mil) 和 α 甘露糖苷酶的体内作用 llx (MX)。 Mil/MX 双无效小鼠胚胎未显示复杂的 A/- 型聚糖， 表明 Mil 和 MX 一起负责复杂型 N-聚糖的合成。军用/MX 双空基因在出生后不久就会致命。根据这些发现，具体目标是： (1) 使用条件 Mil/MX 双无效测定体内基于 A/-聚糖的 L-选择素配体活性 老鼠。内皮细胞和造血细胞特异性 Mil/MX 双敲除 (Tie2-Cre:Mllftaxp/floxp/MX-/-) 将生成并分析高内皮小静脉 (HEV) 中的 L-选择素配体活性，(2) 定义 Sfrs 和 Anxal 的碳水化合物结合活性，(3) 以确定是否发生靶向凋亡 踝关节束缚。 Anxal 已被鉴定为肿瘤特异性内皮标志物，我们鉴定了 IF 肽， 它优先与 Anxal 结合。因此，我们将开发一种靶向细胞凋亡的方法 使用 IF-肽对小鼠肿瘤脉管系统的内皮细胞进行分析，并且 (4) 确定 Anxal 是否 在内皮表面表达促进血管生成并确定潜在机制 Anxal 依赖性内皮细胞激活，包括与硫酸乙酰肝素的结合。 这些研究将为更好地理解潜在机制提供信息 碳水化合物依赖性癌症转移并有助于开发新的治疗策略 上皮癌。