Molecular targeting of PPAR-delta in colon cancer

结肠癌中 PPAR-δ 的分子靶向

基本信息

批准号：
8657861
负责人：
Imad Shureiqi
金额：
$ 30.85万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8657861
关键词：
Agonist Angiogenic Factor Azoxymethane Binding Biological Assay Bladder Blood Vessels Breast Cancer Etiology Cancer cell line Cells Cessation of life Clinical Colon Carcinoma Colonic Adenoma Colonic Neoplasms Colonic Polyps Data Development Dinoprostone Disease Down-Regulation Dyslipidemias Electrophoretic Mobility Shift Assay Ensure Enzyme-Linked Immunosorbent Assay Epithelial Cells Event Extracellular Fluid Fluorescein Future Genetic Genetic Transcription Germ Lines Goals Growth Growth Factor Overexpression HCT116 Cells HT29 Cells Human Image Immunohistochemistry In Vitro Incidence Interleukin-8 Intervention Intestines Isothiocyanates Knock-out Label Lead Lectin Ligands Liposomes Liver Longitudinal Studies Luciferases Lung Magnetic Resonance Imaging Malignant Neoplasms Malignant neoplasm of pancreas Metastatic Neoplasm to the Liver Modeling Molecular Molecular Target Morbidity - disease rate Mus Neoplasm Metastasis Nude Mice Null Lymphocytes PECAM1 gene PPAR delta Perfusion Pharmaceutical Preparations Pharmacologic Substance Polymerase Chain Reaction Pre-Clinical Model Process Production Protein Secretion Proteins Publishing Relative (related person)Reporting Research Resistance Role Safety Signal Pathway Small Interfering RNA Staging Stromal Cells System Testing Theoretical model Therapeutic Intervention Therapy Evaluation Time Tumor Angiogenesis United States Up-Regulation VEGFA gene Vascular Endothelial Growth Factors Xenograft procedure adenoma angiogenesis base cancer cell cancer therapy chemotherapy chromatin immunoprecipitation clinical efficacy colon cancer cell line density improved in vivo inhibitor/antagonist knock-down liver xenograft mRNA Expression mortality mouse model neoplastic cell novel strategies overexpression pre-clinical promoter public health relevance success therapeutic target treatment strategy tumor tumor growth tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Our long-term goal is to help develop new molecularly targeted colon cancer treatments. Angiogenesis is critical for colonic tumorigenesis. The peroxisome proliferator-activated receptor-delta (PPAR-d) is upregulated in human colon cancer. Published data are discordant on the effects of germ line PPAR-d genetic knock out (KO) on intestinal tumorigenesis in APCMin mice and the effects of PPAR-d agonist on vascular endothelial growth factor (VEGF) expression in cancer cell lines and intestinal adenomas of APCMin mice. The impact of PPAR-d upregulation in cancer cells on angiogenesis and tumorigenesis remains to be determined. Our preliminary data show that targeted intestinal PPAR-d KO profoundly inhibited azoxymethane-induced murine colonic tumorigenesis and VEGF expression; PPAR-d re-expression in PPAR-d null HCT-116 (PPAR-d-KO) cells restored their ability to form liver metastases and enhanced VEGF and interleukin-8 (IL-8) expression; and liposomal PPAR-d siRNA inhibited tumorigenesis and PPAR-d, VEGF, and IL-8 expression in HCT-116 in mice. We hypothesize that PPAR-d overexpression in colon cancer cells upregulates VEGF and IL-8 expression to promote tumor angiogenesis and tumorigenesis. Aim 1 is to determine if PPAR-d specific expression in colon cancer cells promotes tumorigenesis and angiogenesis and upregulates VEGF and IL-8 expression, by examining the effects of PPAR-d re-expression in PPAR-d-KO cells on angiogenesis markers (e.g. CD31 immunohistochemistry; FITC-lectin assay), tumorigenesis (liver and lung metastasis formation), and VEGF and IL-8 expression (mRNA by quantitative RT-PCR, protein by ELISA) in nude mice. Aim 2 is to determine whether PPAR-d knockdown via systemic delivery of liposomal PPAR-d siRNA to colon cancer cells in vivo is sufficient to inhibit tumorigenesis and angiogenesis and downregulate VEGF and IL-8 expression, by examining the effects of liposomal PPAR-d siRNA on angiogenesis, tumorigenesis (liver and lung metastasis formation by HCT-116 and HT-29 cells in nude mice), and PPAR-d, VEGF, and IL-8 expression. Aim 3 is to determine VEGF role in PPAR-d promotion of angiogenesis and tumorigenesis, by examining in nude mice the effects of VEGF overexpression in PPAR-d-KO cells on angiogenesis and tumorigenesis; assessing the effects of VEGF liposomal-siRNA knockdown on angiogenesis and tumorigenesis promoted by PPAR-d re-expression in PPAR-d-KO cells; and comparing the effects of PPAR-d and VEGF downregulation (via liposomal siRNA) on angiogenesis and liver and lung metastasis formation by HCT-116 and HT-29 cells. Aim 4 is to determine IL-8 role in PPAR-d promotion of angiogenesis and tumorigenesis, as done for VEGF in specific Aim 3. In Aims 3 and 4, we will also investigate if PPAR-d binds to the VEGF and IL-8 promoters to enhance their transcription, using VEGF and IL-8 promoter-luciferase deletion construct assays, EMSA, and ChIP/real-time PCR in HCT- 116 cells with wild-type PPAR-d expression, PPAR-d overexpression, or PPAR-d-KO. Confirmation of the tested hypothesis could lead to developing PPAR-d-targeted therapy to inhibit tumorigenesis.

描述（由申请人提供）：我们的长期目标是帮助开发新的分子靶向结肠癌治疗。血管生成对于结肠肿瘤发生至关重要。在人类结肠癌中，过氧化物酶体增殖物激活的受体 - 戴尔（PPAR-D）上调。公开的数据对种系PPAR-D遗传敲除（KO）对APCMIN小鼠的肠道肿瘤发生的影响以及PPAR-D激动剂对APCMIN小鼠癌细胞系和肠道腺瘤的血管内皮生长因子（VEGF）表达的影响。 PPAR-D上调在癌细胞中对血管生成和肿瘤发生的影响尚待确定。我们的初步数据表明，靶向肠道PPAR-D KO深刻抑制了偶氮甲烷诱导的鼠结肠肿瘤发生和VEGF表达。 PPAR-D在PPAR-D NULL HCT-116（PPAR-D-KO）细胞中的重新表达恢复了它们形成肝转移并增强VEGF和Interleukin-8（IL-8）表达的能力；脂质体PPAR-D siRNA抑制小鼠HCT-116中的肿瘤发生和PPAR-D，VEGF和IL-8表达。我们假设结肠癌细胞中的PPAR-D过表达上调VEGF和IL-8表达以促进肿瘤血管生成和肿瘤发生。目的1是通过检查PPAR-D重新表达在PPAR-D-KO细胞对血管生成标记物中PPAR-D重新表达对血管生成标记物中的影响，确定PPAR-D特异性表达是否促进了肿瘤发生和血管生成并上调VEGF和IL-8表达。裸鼠和IL-8表达（通过定量RT-PCR，蛋白质的mRNA）在裸鼠中。目的2是确定在体内通过全身传递脂质体PPAR-D siRNA向结肠癌细胞的PPAR-D敲除足以抑制肿瘤的发生和血管生成，血管生成，并下调VEGF和IL-8表达，通过检查脂质体PPAR-DNNA对脂质体PPAR-sirna对血管生成，limorigiS的作用（limorigiss）的影响（liposomal ppar-intrasewosion 3）裸鼠）和PPAR-D，VEGF和IL-8表达。 AIM 3是通过在裸鼠中检查VEGF过表达对PPAR-D-KO细胞对血管生成和肿瘤发生的影响，以确定VEGF在PPAR-D促进血管生成和肿瘤发生的作用。评估VEGF脂质体 - 丝Na敲低对PPAR-D-KO细胞中PPAR-D重新表达促进的血管生成和肿瘤发生的影响；并比较PPAR-D和VEGF下调（通过脂质体siRNA）对HCT-116和HT-29细胞的血管生成以及肝脏和肺转移形成的影响。 Aim 4 is to determine IL-8 role in PPAR-d promotion of angiogenesis and tumorigenesis, as done for VEGF in specific Aim 3. In Aims 3 and 4, we will also investigate if PPAR-d binds to the VEGF and IL-8 promoters to enhance their transcription, using VEGF and IL-8 promoter-luciferase deletion construct assays, EMSA, and ChIP/real-time PCR in HCT- 116 cells with野生型PPAR-D表达，PPAR-D过表达或PPAR-D-KO。对测试假设的确认可能会导致靶向PPAR-D靶向疗法抑制肿瘤发生。