Role of Heparan Sulfate-Degrading Sulfatases in Pancreatic Adenocarcinomas

硫酸乙酰肝素降解硫酸酯酶在胰腺腺癌中的作用

• 批准号: 7268129
• 负责人: STEVEN D ROSEN
• 金额: $ 19.89万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7268129
• 关键词: Adenocarcinoma Cell; Attention; Binding; Biological; Blocking Antibodies; Cell Line; Cell surface; Cells; Coculture Techniques; Development; Disease; Dominant-Negative Mutation; Drug Delivery Systems; Embryo; Embryonic Development; Enzymes; Erinaceidae; GAG Gene; Glucosamine; Growth; Growth Factor; Heparan Sulfate Proteoglycan; Heparitin Sulfate; Human; In Vitro; Incidence; Ligands; Malignant Neoplasms; Malignant neoplasm of pancreas; Modification; Motivation; Nature; Nude Mice; Organ; Pancreas; Pancreatic Adenocarcinoma; Pathway interactions; Pharmaceutical Preparations; Proteins; Publishing; Radiation; Rate; Relative (related person); Research; Resistance; Review Literature; Role; Sampling; Signal Pathway; Signal Transduction; Stem cells; Subfamily lentivirinae; Sulfatases; Transcript; Tumorigenicity; Work; base; beta catenin; cancer cell; cancer stem cell; cell growth; cell transformation; data mining; extracellular; inhibitor/antagonist; interest; morphogens; mortality; mutant; notch protein; novel; pancreatic neoplasm; receptor; self-renewal; small hairpin RNA; small molecule; stem; sulfation; tumor; tumor progression

DESCRIPTION (provided by applicant): Pancreatic adenocarcinoma is one of the most deadly malignancies in human for which there are very limited treatment options. There is great interest in the possible role of embryonic signaling pathways in the initiation and progression of this cancer and others. Among these pathways is canonical Wnt signaling, which is involved in the proliferation and self-renewal of stem/progenitor cells and has been speculated to have a similar role in cancer stem cells. The presence of activated beta-catenin in pancreatic tumor samples suggests that 30-40% of these tumors manifest Wnt signaling. Moreover, Matthias Hebrok and colleagues at UCSF have discovered that active Wnt signaling is required for the growth of several pancreatic adenocarcinoma cell lines in vitro. Our exploratory R21 project proposes to investigate the role of two novel sulfatases, called HSulf-1 and HSulf-2, in Wnt signaling within pancreatic adenocarcinomas and their contributions to the growth and tumorigenicity of these cancer cells. The Sulfs are extracellular enzymes which act on heparan sulfate proteoglycans (HSPGs) to remove a specific sulfation modification (glucosamine-6-O-sulfation). One known biological activity of the Sulfs is to modulate the interaction of Wnt ligands with HSPGs and to potentiate the ability of the Wnts to activate their signal transduction receptors. Our preliminary studies have found the expression of SULF1 or SULF2 transcripts in 23 of 24 pancreatic adenocarcinoma cell lines and the presence of Sulf-1 or Sulf-2 protein in 4/4 of these cell lines and 3/5 of pancreatic tumor samples. Expression of a dominant negative form (enzymatically-inactive mutant) of either Sulf-1 or Sulf-2 in 3 out of 4 of these lines resulted a reduction in Wnt signaling. Furthermore, co-culturing the same three cell lines in the presence of inactive Sulf protein produced a parallel reduction in Wnt signaling and cell growth in vitro. The proposed research will employ lentivirus-transduced shRNA expression to silence one or both Sulfs in representative pancreatic adenocarcinoma cell lines. Our Aims are: 1) To determine the effects of Sulf silencing on the growth and Wnt signaling of these cells in vitro; and 2) To determine the effects of Sulf silencing on the ability of the cells to form tumors in nude mice. Positive results from these studies should stimulate considerable interest in the Sulfs as possible targets (for small molecules or function-blocking antibodies) for the treatment of pancreatic adenocarcinoma in humans.

描述（由申请人提供）：胰腺腺癌是人类最致命的恶性肿瘤之一，治疗方案非常有限。人们对胚胎信号通路在该癌症和其他人的启动和发展中的可能作用引起了极大的兴趣。这些途径包括规范的Wnt信号传导，它参与了茎/祖细胞的增殖和自我更新，并被推测在癌症干细胞中具有相似的作用。胰腺肿瘤样品中活化的β-catenin的存在表明，这些肿瘤中有30％表现出Wnt信号。此外，UCSF的Matthias Hebrok及其同事发现，在体外几种胰腺腺癌细胞系生长需要活跃的Wnt信号传导。我们的探索性R21项目提议研究两种新型硫酸酶（称为HSULF-1和HSULF-2）在胰腺腺癌内Wnt信号传导中的作用，及其对这些癌细胞生长和肿瘤性的贡献。硫是作用于硫酸乙酰肝素蛋白聚糖（HSPG）的细胞外酶，以去除特定的硫酸化修饰（葡萄糖胺-6-O-硫化）。硫的一种已知的生物学活性是调节Wnt配体与HSPG的相互作用，并增强WNT激活其信号转导受体的能力。我们的初步研究发现，在24个胰腺腺癌细胞系中有23种Sulf1或Sulf2转录本在这些细胞系中的4/4和胰腺肿瘤样品中的3/5中，Sulf-1或Sulf-2蛋白的存在。在这些线中的4个中，有4个中有3个中的Sulf-1或Sulf-2的显性阴性形式（酶活性突变体）的表达导致Wnt信号的降低。此外，在存在非活性硫蛋白的情况下共同培养相同的三个细胞系可在体外产生Wnt信号传导和细胞生长的平行降低。拟议的研究将采用慢病毒转移的shRNA表达，使代表性胰腺腺癌细胞系中的一种或两种硫沉默。我们的目的是：1）确定硫沉默对这些细胞在体外的生长和Wnt信号传导的影响； 2）确定硫沉默的影响对细胞在裸鼠中形成肿瘤的能力。这些研究的阳性结果应刺激对硫的相当大的兴趣（对于小分子或功能阻断抗体），以治疗人类胰腺腺癌。