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15-LOX-1 regulation of resolving generation to modulate colon cancer

15-LOX-1 调节分解生成以调节结肠癌

基本信息

批准号：
10301423
负责人：
Imad Shureiqi
金额：
$ 11.54万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10301423
关键词：
ALOX15 gene Affect Apoptosis Arachidonate 15-Lipoxygenase Arachidonate 5-Lipoxygenase Attenuated Azoxymethane Breeding Cancerous Cell Differentiation process Cell Proliferation Cell Respiration Cells Chemoprevention Chemopreventive Agent Clinical Colitis Colon Carcinoma Colorectal Cancer Cyclin D1 Data Development Diet Diffusion Down-Regulation Enzyme Immunoassay Enzymes Epithelial Cells Event FDA approved Fish Oils Formulation Future Generations Genes Genetic Genetic Transcription Goals Health Human Hyperlipidemia Incidence Integration Host Factors Interleukin-1 beta Intervention Intestines Knock-out Knockout Mice LOX gene Length Lentivirus Lentivirus Vector Leukocytes Lipoxygenase 1 Measures Mediating Messenger RNA Metabolism Molecular Molecular Target Mus Mutation Omega-3 Fatty Acids Organoids Pathway interactions Patients Pharmacologic Substance Phase Preventive Production Proteins Quantitative Reverse Transcriptase PCR Regulation Research Shapes Signal Pathway Signal Transduction TNF gene Testing Transgenic Organisms Work base beta catenin c-myc Genes cancer chemoprevention colon cancer prevention colon tumorigenesis colonic crypt dietary supplements gain of function improved intestinal epithelium liquid chromatography mass spectrometry loss of function mouse model novel premalignant prevent reconstitution small hairpin RNA tumor tumorigenesis

项目摘要

Our long-term goal is to develop novel interventions to prevent colorectal tumorigenesis (CRT). The effects of fish oil and its omega-3 fatty acid derivatives DHA and EPA on CRT are controversial: some studies show promotion and others suppression. We found in preliminary studies that fish oil promoted while EPA inhibited colitis-associated CRT. Whether these differences are applicable to sporadic CRT is unknown. This is clinically important because DHA and EPA are widely used as dietary supplements and FDA-approved treatments. 15- lipoxygenase-1 (15-LOX-1) is a critical enzyme for DHA and EPA oxidative metabolism to generate resolvins and their precursors (e.g., 18-HEPE and RvEs, 17-HDHA and RvEs). 15-LOX-1 and resolvins suppress im- portant pathways (e.g., TNF-α, IL-1β) that potentiate aberrant Wnt/beta-catenin (Wnt/B-catenin) signaling, which is a driver of CRT. 15-LOX-1 expression is commonly lost early during human CRT. Whether 15-LOX-1 expression loss reduces resolvin production from DHA and EPA and reduces the effects of DHA and EPA on CRT is unknown. Our preliminary data show that intestinally targeted 15-LOX-1 expression (15-LOX-1-Gut mice) enhanced resolvin production, suppressed APC mutation-driven B-catenin activation and CRT, attenuat- ed DHA-induced CRT promotion, and enhanced EPA-induced CRT suppression. We hypothesize that 15-LOX- 1 expression in colonic epithelial cells is critical for DHA and EPA to generate resolvins and modulate Wnt/B- catenin signaling and CRT. Aims 1 and 2 will determine the effects of 15-LOX-1 gain (aim 1) and loss of func- tion (aim 2) in colonic epithelial cells on resolvin generation from DHA and EPA, CRT, and B-catenin signaling. For aim 1, we will use 15-LOX-1-Gut mice. For aim 2, we will breed mice with intestinal 12-S-LOX transgenic expression with 12/15-LOX knockout mice to target 12-S-LOX reconstitution to intestinal epithelial cells. In both aims, mice will be fed control, DHA, or EPA diet and treated with AOM to induce CRT. Groups will be com- pared for resolvins and their precursors, CRT, cell proliferation and apoptosis, TNF-α, IL-1β, and activated B- catenin levels, and Wnt/B-catenin target gene (c-Myc, Cyclin D1, Axin2) mRNA levels. We will also evaluate the effect of 15-LOX-1 gain of function (aim 1) and downregulation (aim 2) in human colonic cancerous and normal organoids isolated from patients' colonic crypts and cultured with various concentrations of EPA and DHA on resolvin generation, cell proliferation, cell differentiation, and Wnt/B-catenin signaling. Aim 3 will de- termine the temporal effects of 15-LOX-1 expression in colonic epithelial cells and in leukocytes on resolvin generation and CRT. 15-LOX-1 transgenic expression will be targeted to intestinal epithelial cells and induced at either initiation or progression phases of AOM-induced CRT (subaim 3A) or targeted to leukocytes prior to AOM-induced CRT (subaim 3B). Mice will be fed DHA or EPA diets and compared for end points described for aims 1 and 2. Our findings will direct efforts to develop interventions to prevent CRT based on understanding of the host factors (e.g., 15-LOX-1) that affect DHA and EPA modulation of CRT.

我们的长期目标是开发新的干预措施来预防结直肠肿瘤发生（CRT）。的影响鱼油及其 omega-3 脂肪酸衍生物 DHA 和 EPA 对 CRT 的影响存在争议：一些研究表明晋升和他人压制。我们在初步研究中发现，鱼油具有促进作用，而 EPA 具有抑制作用结肠炎相关的 CRT。这些差异是否适用于偶发性 CRT 尚不清楚。这是临床上重要的是因为 DHA 和 EPA 被广泛用作膳食补充剂和 FDA 批准的治疗方法。 15- 脂氧合酶-1 (15-LOX-1) 是 DHA 和 EPA 氧化代谢产生分解素的关键酶及其前体（例如 18-HEPE 和 RvE、17-HDHA 和 RvE）。 15-LOX-1 和解析素抑制 im- 增强异常 Wnt/β-连环蛋白 (Wnt/B-连环蛋白) 信号传导的重要途径（例如 TNF-α、IL-1β），这是CRT的驱动程序。 15-LOX-1 表达通常在人类 CRT 早期丢失。是否15-LOX-1 表达缺失会减少 DHA 和 EPA 分解素的产生，并降低 DHA 和 EPA 对 CRT 未知。我们的初步数据表明，肠道靶向 15-LOX-1 表达（15-LOX-1-Gut 小鼠）增强了消退素的产生，抑制了 APC 突变驱动的 B-连环蛋白激活和 CRT，减弱了 ed DHA 诱导的 CRT 促进，并增强 EPA 诱导的 CRT 抑制。我们假设 15-LOX- 结肠上皮细胞中的 1 表达对于 DHA 和 EPA 生成分解素和调节 Wnt/B- 至关重要连环蛋白信号传导和 CRT。目标 1 和 2 将确定 15-LOX-1 增益（目标 1）和功能丧失的影响结肠上皮细胞中 DHA 和 EPA、CRT 和 B-连环蛋白信号转导生成解析素的影响（目标 2）。对于目标 1，我们将使用 15-LOX-1-Gut 小鼠。对于目标 2，我们将培育肠道 12-S-LOX 转基因小鼠使用 12/15-LOX 敲除小鼠进行表达，以将 12-S-LOX 重建靶向肠上皮细胞。在两者中为了达到这个目的，小鼠将被喂食对照、DHA 或 EPA 饮食，并用 AOM 治疗以诱导 CRT。团体将由与消解素及其前体、CRT、细胞增殖和凋亡、TNF-α、IL-1β 和活化的 B- 连环蛋白水平和 Wnt/B-连环蛋白靶基因（c-Myc、Cyclin D1、Axin2）mRNA 水平。我们也会评估 15-LOX-1 功能增强（目标 1）和下调（目标 2）对人类结肠癌和结肠癌的影响从患者结肠隐窝中分离出的正常类器官，并与不同浓度的 EPA 和 DHA 对分解素生成、细胞增殖、细胞分化和 Wnt/B-连环蛋白信号传导的影响。目标 3 将消除终止结肠上皮细胞和白细胞中 15-LOX-1 表达对 resolvin 的时间影响一代和 CRT。 15-LOX-1转基因表达将靶向肠上皮细胞并诱导在 AOM 诱导的 CRT (subaim 3A) 的起始或进展阶段或在之前针对白细胞 AOM 诱导的 CRT (subaim 3B)。小鼠将被喂食 DHA 或 EPA 饮食，并比较以下描述的终点目标 1 和 2。我们的研究结果将指导我们在理解的基础上制定预防 CRT 的干预措施影响 CRT 调节 DHA 和 EPA 的宿主因子（例如 15-LOX-1）。