Carbohydrate- Dependent Epithelial Cancer Metastasis

有充分的文献证明，上皮细胞的恶性转化与结构性炎症有关。 细胞表面碳水化合物的变化。许多临床研究已经将那些与癌症相关的 碳水化合物抗原，临床预后差，包括转移。碳水化合物依赖 已经观察到癌症转移，但是该过程的潜在机制尚未确定。 我们先前鉴定了IELLQAR（l-肽），其模拟抗Lewis A抗体的表位， 筛选肽展示噬菌体文库。当将l-肽注射到野生型小鼠中时， 抑制sLex-dependent黑色素瘤肺定植。然而，肺定植也发生在突变体中， 缺乏E-和P-选择素和L-肽的小鼠抑制了定植，排除了 在这个过程中选择。这些发现表明l-肽干扰定殖确实 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）和膜联蛋白AI（Anxal）。 PI还研究了高尔基体加工α-甘露糖苷酶II（Mil）和甘露糖苷酶的体内作用 llx（墨西哥）。Mil/MX双无效小鼠胚胎显示无复合A型/-聚糖， 表明Mil和MX一起负责复合型N-聚糖的合成。密尔/MX 出生后不久，双无效是致命的。根据这些调查结果，具体目标是： (1)使用条件性Mil/MX双无效测定体内基于N-聚糖的L-选择素配体活性 小鼠内皮细胞和造血细胞特异性Mil/MX双敲除（Tie 2-Cre：Mil ftaxp/Mil ftaxp/MX-/-） 将产生并分析高内皮微静脉（HEV）中的L-选择素配体活性，（2）至 确定Sfrs和Anxal的碳水化合物结合活性，（3）确定是否发生靶向凋亡， Anxal绑定。Anxal已被鉴定为肿瘤特异性内皮标志物，我们鉴定了IFpeptide， 其优先结合Anxal。因此，我们将开发一种靶向细胞凋亡的方法， 使用IF-肽的小鼠肿瘤血管内皮细胞，和（4）确定Anxal 在内皮细胞表面表达促进血管生成，并确定潜在的机制 血管内皮依赖性内皮细胞活化，包括与硫酸乙酰肝素的结合。 这些研究将为更好地了解其机制提供信息 碳水化合物依赖性癌症转移，并帮助开发新的治疗策略， 上皮癌

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.