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：
9187612
负责人：
Imad Shureiqi
金额：
$ 36.6万
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：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9187612
关键词：
ALOX15 gene Affect Apoptosis Arachidonate 15-Lipoxygenase Arachidonate 5-Lipoxygenase Attenuated Azoxymethane B cell differentiation Breeding Cancerous Cell Differentiation process Cell Proliferation Cell Respiration Cells Chemoprevention Chemopreventive Agent Colitis Colon Carcinoma Colorectal Cancer Cyclin D1 Data Development Diet Diffusion Down-Regulation Enzyme Immunoassay Enzymes Epithelial Cells Event FDA approved Figs - dietary Fish Oils Formulation Future Generations Genetic Goals Health Human Hyperlipidemia Incidence Integration Host Factors Interleukin-1 beta Intervention Intestines Knock-out Knockout Mice LOX gene Length 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 Premalignant Preventive Production Proteins Regulation Research Shapes Signal Pathway Signal Transduction Staging Subfamily lentivirinae TNF gene Testing Transgenic Organisms Work base beta catenin c-myc Genes cancer chemoprevention colon tumorigenesis colonic crypt dietary supplements feeding gain of function improved liquid chromatography mass spectrometry loss of function mouse model novel 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.

我们的长期目标是制定新的干预措施，以防止结直肠肿瘤发生（CRT）。效果 CRT上的鱼油及其Omega-3脂肪酸衍生物DHA和EPA是有争议的：一些研究表明晋升和其他抑制。我们在初步研究中发现，鱼油促进了EPA抑制结肠炎相关的CRT。这些差异是否适用于零星CRT是未知的。这是临床上的重要的是因为DHA和EPA被广泛用作饮食补充剂和FDA批准的治疗方法。 15- Lipoxyganase-1（15-LOX-1）是DHA和EPA氧化物代谢的关键酶，产生溶异胞菌素及其前体（例如18-Hepe和Rves，17-HDHA和RVE）。 15-LOX-1和Resolvins抑制Im- 潜在的异常Wnt/beta-catenin（Wnt/b-catenin）信号传导潜在的异常Wnt/beta-catenin（例如TNF-α，IL-1β）的途径（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期间丧失。是否15-lox-1 表达损失减少了DHA和EPA的解决产生，并降低了DHA和EPA对 CRT是未知的。我们的初步数据表明，靶向15-lox-1表达（15-lox-1 gut）小鼠）增强了分解产生，抑制APC突变驱动的B-catenin激活和CRT，attenuat- ED DHA诱导的CRT促进，并增强了EPA诱导的CRT抑制。我们假设15-lox- 1在结肠上皮细胞中的表达对于DHA和EPA产生分辨率和调节Wnt/b-至关重要 Catenin信号传导和CRT。目标1和2将确定15-LOX-1增益（AIM 1）的影响和弹性损失结肠上皮细胞中的TION（AIM 2）在DHA和EPA，CRT和B-catenin信号传导中产生的溶质产生。对于AIM 1，我们将使用15-LOX-1螺旋小鼠。对于AIM 2，我们将用肠道12-S-lox转基因繁殖小鼠用12/15-lox基因敲除小鼠的表达，以靶向12-S-Lox重构对肠上皮细胞。在这两个中目标，小鼠将得到控制，DHA或EPA饮食，并用AOM治疗以诱导CRT。小组将是甲状腺素及其前体，CRT，细胞增殖和凋亡，TNF-α，IL-1β和活化的B- Catenin水平和Wnt/B-catenin靶基因（C-MYC，Cyclin D1，Axin2）mRNA水平。我们还将评估人类结肠取消和从患者的结肠加密群中分离出的正常器官，并用各种浓度的EPA培养 DHA在分解生成，细胞增殖，细胞分化和Wnt/b-catenin信号传导上。 AIM 3将脱离终止15-LOX-1表达在结肠上皮细胞和白细胞中的临时作用对Resolvin 一代和CRT。 15-LOX-1转基因表达将针对肠上皮细胞，并诱导在AOM诱导的CRT（subaim 3a）的主动性或进展阶段，或针对白细胞 AOM诱导的CRT（Subaim 3b）。小鼠将被喂食DHA或EPA饮食，并比较描述的终点目标1和2。我们的发现将指导努力制定干预措施，以防止基于理解的CRT 影响CRT的DHA和EPA调制的宿主因子（例如15-LOX-1）。