Specific Targets for Pancreatic Cancer Therapy

胰腺癌治疗的具体目标

• 批准号: 7489979
• 负责人: Ramzi M. Mohammad
• 金额: $ 22.91万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7489979
• 关键词: Animal Model; Apoptosis; Apoptotic; Avidity; BCL-2 Protein; BCL2 gene; BH3 peptide; Binding; Bioinformatics; Biological; Biological Assay; Cancer cell line; Cell Death; Cell Line; Cell Survival; Cells; Cisplatin/Gemcitabine; Clinical; Data; Data Set; Development; Drug resistance; Drug usage; Enzymes; Epidermal Growth Factor Receptor; Equipment and supply inventories; Etiology; Event; Family member; Gelatinase B; Generations; Genes; Genetic Transcription; Genistein; Growth; Heterodimerization; Human; Hybrids; In Vitro; Induction of Apoptosis; Inhibitory Concentration 50; Label; Lead; MCL1 protein; Malignant Neoplasms; Malignant neoplasm of pancreas; Mitochondria; Modeling; Molecular; Mus; NF-kappa B; Neoplasm Metastasis; Newly Diagnosed; Pathway interactions; Peptides; Pharmaceutical Preparations; Play; Protein Family; Protein Overexpression; Proteins; Publishing; Radio; Rate; Research Personnel; Resistance; Role; SCID Mice; Solid Neoplasm; Standards of Weights and Measures; System; Testing; United States; Vascular Endothelial Growth Factors; Weight; Western Blotting; Work; Xenograft Model; Yeasts; angiogenesis; cDNA Library; cancer cell; cancer therapy; caspase-3; celecoxib; chemotherapy; clinical application; cyclooxygenase 2; design; enzyme activity; gemcitabine; gene repression; human MCL1 protein; improved; in vivo; inhibitor/antagonist; killings; mimetics; mortality; novel; novel therapeutics; pancreatic neoplasm; pre-clinical; pro-apoptotic protein; programs; response; small molecule; subcutaneous; tumor; tumor progression; two-dimensional

DESCRIPTION (provided by applicant): Pancreatic neoplasia is the fifth most common cancer in the United States with >28,000 newly diagnosed cases per year with an annual mortality rate of greater than 99%. Its etiology is largely unknown and no curative treatment is presently available. In pancreatic cancer, several important survival molecules such as EGFR, NF-kB, COX-2, and Bcl-xL are highly activated. NF-kB regulates many genes involved in tumor progression, survival, angiogenesis and invasion such as Bcl-xL, COX-2, VEGF, MMP-9 and uPAR. High expression of anti-apoptotic Bcl-xL protein promotes cell survival and plays a key role in pancreatic cancer progression leading to drug resistance. Its expression was found in 90% of pancreatic cancer. The overexpression of the cyclooxygenase-2 (COX-2) enzyme in pancreatic cancer has also been shown to contribute to growth, metastasis, and chemoresistance. Therefore, inhibitions of NF-kB, Bcl-2/Bcl-xL, and COX-2 should serve as a novel treatment strategy for pancreatic cancer. Our co-investigator has designed non-peptide, drug-like, cell permeable potent small molecule inhibitors (SMI) that bind to the pocket of Bcl-2 and Bcl-xL and block/disrupt their anti-apoptotic function. These SMI were tested in an in vitro binding assay and found to be potent inhibitor of the binding of Bak BH3 peptide to Bcl-2 and Bcl-xL. We have also shown that genistein inactivates NF-kB and down regulates Bcl-xL, VEGF, MMP-9, and uPAR by transcriptional inactivation. Furthermore, the inhibition of COX-2 activity and Akt/NF-kB by celecoxib could be potentially useful in killing pancreatic cancer cells. We have found that human pancreatic cancer cell lines are notable for high expression of Bcl-XL, NF-kB, and COX-2. Therefore, we hypothesize that targeting the Bcl-2/Bcl-xL survival pathway by SMI and inactivating NF-kB-induced Bcl-2/BcI-xL/COX-2 generation by genistein and celecoxib could be novel therapeutic strategies for the treatment of pancreatic cancer. Of importance to this work is that NF-kB transcriptionally regulates the expression of Bcl-xL, hence inactivation of NF-kB by genistein and treatment with SMI will lead to decreased Bcl-xL. Moreover, the addition of celecoxib may be very useful for inducing pancreatic cancer cell death by inhibiting NF-kB as well as transcription and enzyme activity of COX-2. Our Specific Aims are: 1) To investigate the molecular mechanisms by which SMI promote apoptosis in human pancreatic cancer cell lines, 2) To determine anti-tumor activity of SMI using human pancreatic cancer xenograft model, 3) To determine whether transcriptional repression of Bcl-xL by genistein will synergize SMI-induced apoptosis in vitro and in vivo, and 4) To determine the effects of inhibition of COX-2 by celecoxib on Akt/NF-kB activation in genistein-treated and SMI-treated pancreatic cancer cells in vitro and in SCID mouse xenograft models. Targeting Bcl-2/Bcl-xL, NF-kB and COX-2 could be novel therapeutic treatment of pancreatic cancer.

描述（由申请人提供）：胰腺肿瘤是美国第五大常见癌症，每年有> 28，000例新诊断病例，年死亡率大于99%。 其病因在很大程度上尚不清楚，目前尚无治愈性治疗方法。 在胰腺癌中，几种重要的存活分子如EGFR、NF-kB、考克斯-2和Bcl-xL高度活化。 NF-kB调节许多参与肿瘤进展、存活、血管生成和侵袭的基因，如Bcl-xL、考克斯-2、VEGF、MMP-9和uPAR。 抗凋亡Bcl-xL蛋白的高表达促进细胞存活，并在胰腺癌进展中发挥关键作用，导致耐药性。 在90%的胰腺癌中有表达。 胰腺癌中环氧合酶-2（考克斯-2）的过度表达也被证明有助于生长、转移和化学抗性。 因此，NF-kB、Bcl-2/Bcl-xL和考克斯-2的抑制应作为胰腺癌的新治疗策略。 我们的合作研究者设计了非肽类、药物样、细胞可渗透的有效小分子抑制剂（SMI），其结合Bcl-2和Bcl-xL的口袋并阻断/破坏其抗凋亡功能。 在体外结合测定中测试这些SMI，发现它们是巴克BH 3肽与Bcl-2和Bcl-xL结合的有效抑制剂。 我们还发现染料木黄酮通过转录失活使NF-kB失活并下调Bcl-xL、VEGF、MMP-9和uPAR。 此外，塞来昔布对考克斯-2活性和Akt/NF-kB的抑制可能对胰腺癌细胞的杀伤有潜在的作用。 我们发现人胰腺癌细胞系中Bcl-XL、NF-κ B和考克斯-2的高表达是值得注意的。 因此，我们推测，通过SMI靶向Bcl-2/Bcl-xL存活通路和通过genistein和塞来昔布灭活NF-κ B诱导的Bcl-2/Bcl-xL/考克斯-2生成可能是治疗胰腺癌的新的治疗策略。 这项工作的重要性是，NF-κ B转录调节Bcl-xL的表达，因此，通过染料木素和SMI治疗NF-κ B的失活将导致Bcl-xL的减少。 此外，塞来昔布的加入可能通过抑制NF-κ B以及考克斯-2的转录和酶活性而对诱导胰腺癌细胞死亡非常有用。 我们的具体目标是：1）研究SMI促进人胰腺癌细胞系凋亡的分子机制，2）使用人胰腺癌异种移植模型确定SMI的抗肿瘤活性，3）确定金雀异黄素对Bcl-xL的转录抑制是否会在体外和体内协同SMI诱导的凋亡，和4）确定塞来昔布抑制考克斯-2对体外和SCID小鼠异种移植模型中染料木素处理和SMI处理的胰腺癌细胞中Akt/NF-kB活化的影响。 以Bcl-2/Bcl-xL、NF-κ B和考克斯-2为靶点的治疗可能成为胰腺癌治疗的新途径。