Cell Penetrating Peptide Inhibitor of FoxM1 in Hepatocellular Carcinoma Treatment

FoxM1 细胞穿膜肽抑制剂在肝细胞癌治疗中的应用

• 批准号: 8018494
• 负责人: Pradip Raychaudhuri
• 金额: $ 27.79万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-25 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8018494
• 关键词: Abnormal Cell; Albumins; Amino Acid Sequence; Amino Acids; Antibodies; Apoptosis; Arginine; Biological Assay; Birth; Boxing; Breeding; Bromodeoxyuridine; Cell Nucleolus; Cell Proliferation; Cell division; Cells; Complementary DNA; Detection; Development; Diethylnitrosamine; E2F1 gene; Enhancers; Exhibits; Frequencies; Future; Gene Expression; Gene Targeting; Genotype; Goals; Growth; HRAS gene; Hepatic; Hepatocyte; Human; Injection of therapeutic agent; Kupffer Cells; Liver; Liver neoplasms; Lung; Lung Neoplasms; MAP Kinase Gene; MAPK Signaling Pathway Pathway; Malignant Neoplasms; Malignant neoplasm of liver; Methods; Mus; Necrosis; Neoplasm Metastasis; Oncogenic; Organ; Peptides; Phenobarbital; Prealbumin; Primary carcinoma of the liver cells; Promoter Regions; Protein p53; Proteins; Recurrence; Reporting; Research Personnel; Resistance; Rosa; Signal Transduction; Simian virus 40; Stimulus; Stromelysin 1; System; Technology; Testing; Time; Transforming Growth Factors; Transgenes; Transgenic Mice; Transgenic Organisms; Translational Research; Tumor Promoters; Tumor Suppressor Proteins; Western Blotting; Wild Type Mouse; angiogenesis; aurora B kinase; base; c-myc Genes; cancer cell; chromatin immunoprecipitation; collagenase 3; cyclin A2; design; effective therapy; hepatic necrosis; human PLK1 protein; in vivo; inhibitor/antagonist; mouse model; neoplastic cell; postnatal; prevent; programs; promoter; recombinase; response; stellate cell; survivin; transcription factor; tumor

DESCRIPTION (provided by applicant): Human hepatocellular carcinoma (HCC) is the fifth most common cancer, yet it is among the most lethal cancers world wide because late detection and high frequency of tumor recurrence render current HCC therapy ineffective. We previously showed that mouse hepatocytes deficient (-/-) in the proliferation-specific Forkhead Box ml (Foxm1) transcription factor are highly resistant to developing Diethylnitrosamine (DEN)/Phenobarbital (PB) induced liver cancer and that FoxM1 transcriptional activity is inhibited by amino acids 26 to 44 from the ARF tumor suppressor. We also developed transgenic (TG) mice in which the Rosa26 promoter was used to drive ubiquitous expression of the human FoxM1b cDNA transgene. We have developed a new mouse model of aggressive metastatic liver cancer. After 33 weeks of DEN/PB exposure, ARF-/- Rosa-26 FoxM1b TG mouse livers are necrotic and develop aggressive HCC that metastasized to the lungs. In Aiml, we propose to further characterize the proliferation, development and metastasis of this aggressive liver cancer and determine whether these hepatic tumors exhibit necrosis and activation of hepatic Kupffer and stellate cells. In preliminary studies, we pharmacologically reduced Foxm1 activity in HCC in vivo by subjecting DEN/PB treated wild type (WT) mice to daily injections of a cell penetrating ARF 24 to 46 peptide. After 4 weeks of this ARF peptide treatment. HCC regions display reduced cell proliferation and angiogenesis with a selective apoptosis of HCC. However, whether this ARF 26-44 peptide is also effective in preventing metastasis of liver cancer to the lung remains to be determined. In Aim2, we propose to test the hypothesis that treatment of ARF -/- Rosa26 FoxM1b TG liver tumors with the cell penetrating ARF 26-44 peptide will inhibit HCC growth and lung metastasis. We have developed a new TG mouse line that conditionally expresses activated H-Ras in postnatal hepatocytes. We will use these mice to examine whether activated H-Ras stimulates progression of DEN induced liver tumors in both Rosa26-FoxM1b TG mice and ARF -/- Rosa26-FoxM1b TG mice and whether the cell penetrating ARF 26-44 peptide is an effective treatment to limit growth and progression of these liver tumors. Completion of the proposed studies will characterize new mouse models of aggressive metastatic liver cancer and determine whether the cell penetrating WT ARF 26-44 peptide is an effective treatment to prevent metastasis of liver cancer to the lung. These studies will facilitate development of a rational design for future translational research in the treatment of human liver cancer with this ARF peptide inhibitor of the FoxM1 transcription factor.

描述（由申请人提供）：人类肝细胞癌（HCC）是第五大最常见的癌症，但它是世界上最致命的癌症之一，因为较晚的检测和肿瘤复发的高频率使得当前的 HCC 治疗无效。我们之前表明，增殖特异性 Forkhead Box ml (Foxm1) 转录因子缺陷 (-/-) 的小鼠肝细胞对二乙基亚硝胺 (DEN)/苯巴比妥 (PB) 诱导的肝癌具有高度抵抗力，并且 FoxM1 转录活性受到 ARF 肿瘤抑制因子的氨基酸 26 至 44 的抑制。我们还开发了转基因 (TG) 小鼠，其中使用 Rosa26 启动子来驱动人类 FoxM1b cDNA 转基因的普遍表达。我们开发了一种新的侵袭性转移性肝癌小鼠模型。 DEN/PB 暴露 33 周后，ARF-/- Rosa-26 FoxM1b TG 小鼠肝脏坏死并发展为侵袭性 HCC，并转移至肺部。在Aiml中，我们建议进一步表征这种侵袭性肝癌的增殖、发展和转移，并确定这些肝脏肿瘤是否表现出肝枯否细胞和星状细胞的坏死和激活。在初步研究中，我们通过每天向 DEN/PB 处理的野生型 (WT) 小鼠注射细胞穿透性 ARF 24 至 46 肽，从药理学角度降低了体内 HCC 中的 Foxm1 活性。经过 4 周的 ARF 肽治疗。 HCC 区域显示细胞增殖和血管生成减少，并伴有 HCC 选择性凋亡。然而，这种ARF 26-44肽是否也能有效预防肝癌肺转移仍有待确定。在 Aim2 中，我们建议测试以下假设：用细胞穿透性 ARF 26-44 肽治疗 ​​ARF -/- Rosa26 FoxM1b TG 肝肿瘤将抑制 HCC 生长和肺转移。我们开发了一种新的 TG 小鼠系，其在出生后肝细胞中条件性表达激活的 H-Ras。我们将使用这些小鼠来检查激活的 H-Ras 是否会刺激 Rosa26-FoxM1b TG 小鼠和 ARF -/- Rosa26-FoxM1b TG 小鼠中 DEN 诱导的肝肿瘤的进展，以及细胞穿透 ARF 26-44 肽是否是限制这些肝肿瘤生长和进展的有效治疗方法。拟议研究的完成将表征侵袭性转移性肝癌的新小鼠模型，并确定细胞穿透性 WT ARF 26-44 肽是否是预防肝癌肺部转移的有效治疗方法。这些研究将有助于为未来利用 FoxM1 转录因子 ARF 肽抑制剂治疗人类肝癌的转化研究制定合理的设计。